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+{"text": "Members of the EGF-CFC family play essential roles in embryonic development and have been implicated in tumorigenesis. The TGF\u03b2 signals Nodal and Vg1/GDF1, but not Activin, require EGF-CFC coreceptors to activate Activin receptors. We report that the TGF\u03b2 signaling antagonist Lefty also acts through an EGF-CFC-dependent mechanism. Lefty inhibits Nodal and Vg1 signaling, but not Activin signaling. Lefty genetically interacts with EGF-CFC proteins and competes with Nodal for binding to these coreceptors. Chimeras between Activin and Nodal or Vg1 identify a 14 amino acid region that confers independence from EGF-CFC coreceptors and resistance to Lefty. These results indicate that coreceptors are targets for both TGF\u03b2 agonists and antagonists and suggest that subtle sequence variations in TGF\u03b2 signals result in greater ligand diversity. TGF\u03b2 family members and their receptors are involved in setting up the left-right body axis early in development. This article clarifies the role of Lefty and elucidates the molecular basis for signaling diversity between the family members The analysis of whole-genome sequences has revealed that most signaling systems consist of multiple ligands that converge on a relatively small set of receptors and pathway-specific transcription factors. In the case of human transforming growth factor-\u03b2 (TGF\u03b2) signaling, 42 TGF\u03b2s converge on seven type I receptors, five type II receptors, and two classes of Smad signal transducers (reviewed in oep) lack all endoderm and most mesoderm, similar to the double mutants for the zebrafish Nodal-related genes cyclops and squint (sqt)  . Moreove mutants . During  mutants .The EGF-CFC protein Cripto is highly overexpressed in human epithelial cancers, such as breast and colon carcinomas , or zebrafish Vg1 (a chimeric molecule containing the Xenopus Activin\u03b2B prodomain fused to the mature domain of zebrafish Vg1) in zebrafish embryos (goosecoid (gsc). Activin\u03b2B, Activin\u03b2A, Sqt, and Vg1-induced ectopic gsc expression in wild-type embryos  expression was only weakly induced , receptors (mouse ActRIIB[KR]/Myc and human Alk4[KR]/Flag), and a coreceptor (mouse Cripto/Flag) in  embryos . Similar embryos , Sqt for embryos A. In con embryos A. Since  embryos , Lefty s embryos B. Since  embryos , these t embryos C. To detep zebrafish embryos and analyzed the ectopic induction of the downstream genes ntl and gsc. Sqt, Vg1, and Activins induced these genes in wild-type embryos, allowing us to determine which chimeric ligands were active. Activins, but not Sqt or Vg1, were active in MZoep mutants, allowing us to test which sequences conferred EGF-CFC coreceptor dependence or independence.The finding that TGF\u03b2 ligands that activate Activin receptors can be grouped into a EGF-CFC coreceptor-dependent class that is susceptible to inhibition by Lefty  and a class that is independent of EGF-CFC proteins and resistant to Lefty (Activin) prompted us to examine which sequences underlie this ligand diversity. We therefore generated chimeras between Activins (EGF-CFC-independent) and Sqt or Vg1 (EGF-CFC-dependent) Figures  and 5. Aoep embryos  were sufficient to confer EGF-CFC independence. Further dissection of this region into loop alone (SqtAct\u03b2B[loop]) or \u03b28 alone (SqtAct\u03b2B[\u03b28]) yielded no or much weaker activity in MZoep mutants as compared with wild-type embryos  and Vg1Act\u03b2B[loop\u03b28] (Vg1 with an Activin\u03b2B loop-\u03b28 region) both induced gsc expression in MZoep mutants with similar efficiencies as in wild-type embryos  and Vg1. Sqt embryos D. These  embryos E and sugAct\u03b2B[loop\u03b28] can signal in an EGF-CFC-independent manner in vivo, suggesting that this chimeric protein might bind to ActRIIB and Alk4 receptors in the absence of EGF-CFC coreceptors. To test this idea, we coexpressed and immunoprecipitated epitope-tagged ligands , receptors (ActRIIB[KR]/Myc and Alk4[KR]/Flag), and a coreceptor (Cripto/Flag) in Xenopus embryos  in Sqt did not enhance activity in MZoep mutants. These results suggest that multiple residues contribute to coreceptor independence, with the type II receptor-binding interface being an essential determinant.An alignment of the loop-\u03b28 region of EGF-CFC-dependent and EGF-CFC-independent TGF\u03b2s A revealson assay . All conoep mutants. Chimeras containing the Sqt Finger 2 domain . The addition of Finger 1 in SAS or ActSqt[Finger1-loop\u03b28] relieved the inhibitory effect of the loop-\u03b28 region of Sqt in wild-type embryos , previous studies have suggested that heterologous Nodal, Lefty, and EGF-CFC proteins have similar activities in zebrafish . For exaLefty1 and left-side specific Lefty2 (\u0394ASELefty2) mutants have been thought to be caused by inappropriate spread of Nodal signaling  fused to an HA-epitope/XhoI fragment (YPYDVPDYALE) and followed by the mature chimeric ligand into pcDNA3 vector. S denotes Sqt; A denotes Activin\u03b2B. The boundaries for Sqt mature ligand domains are as indicated: Finger 1 (Asn263 to Cys325), Heel (Pro326 to Cys358), and Finger 2 . The boundaries for Xenopus Activin\u03b2B mature ligand domains are as indicated: Finger 1 (Cys215 to Cys299), Heel (Pro300 to Cys335), and Finger 2 (Ile336 to Ala370). Full-length chimeras were generated by incorporating the indicated regions into Sqt, Activin\u03b2B, or zebrafish Vg1, which were then subcloned into the pT7TS vector , loop (Phe348 to Ile354), \u03b28 , and \u03b29 (Met357 to Ala370). The Xenopus Activin\u03b2A loop-\u03b28 region sequence is FDRNNNVLKTDIAD . The zebrafish Vg1 loop-\u03b28 region is from Try332 to Asp345. pcDNA3-zebrafish Vg1/HA, pcDNA3-Squint/HA, pCS2-Alk4(KR)/Flag (a kinase-defective mutant of human Alk4 with Lys234 to Arg234 substitution), pCS2-ActRIIB(KR)/Myc (a kinase-defective mutant of mouse ActRIIB with Lys217 to Arg217 substitution), and pCS2-Cripto/Flag have been described elsewhere  Sqt/Activin\u03b2B chimeras were generated by subcloning the prodomain of S vector . The boulsewhere .Xenopus embryos at the one- to two-cell stage were used for injections into the animal pole.Plasmids were linearized and sense strand-capped mRNA was synthesized using the mMESSAGE mMACHINE system . Zebrafish embryos were dechorinated by pronase treatment and injected between the one- and four-cell stage. gsc and ntl  M2Bio dissecting microscope. In situ hybrization was performed as described previously , using R and ntl .LacZ mRNA. The A3-luc reporter DNA construct (25 pg)   was alsoXenopus embryos were harvested at stage 10. For chemical cross-linking of proteins, animal halves were incubated in PBS with 10 mM 3,3\u2032-dithiobis(sulfo-succinimidyl propionate) (DTSSP)  and incubated for 2 h on ice. Coimmunoprecipitation was performed as described previously (eviously . Purifieeviously . Sampleshttp://www.ncbi.nlm.nih.gov/entrez/query.fc-gi?db=Nucleotide) accession numbers for the sequences discussed in this paper are Activin\u03b2A (Q9W6I6), Activin\u03b2B (Q91350), human ActRIIB (P08476), mouse ActRIIB (P27040), rat ActRIIB (P38445), Alk4 (Z22536), BMP7 (P23359), Cripto (P51865), Cryptic (P97766), cyclops (P87358), GDF1 (P20863), goosecoid (P53544), Lefty1 (Q9W6I6), Lefty2 (P57785), mLefty1 (Q64280), Nodal (P43021), no tail (Q07998), one-eyed pinhead (O57516), Pitx2 (P97474), squint (O13144), VEGF-D (P97946), and Vg1 (P09534).The GenBank ("}
+{"text": "Most animals (including humans) show a high level of bilateral symmetry: on the surface, the right side of our body resembles the left. A closer and deeper look, however, reveals an underlying asymmetry. The heart, for example, is on the left side in most humans, and the liver on the right. This left-right asymmetry develops early on in the embryo, and research in the past few years has revealed some of the molecular and cellular mechanisms that establish the left-right axis, which conveys positional information to cells in the growing embryo. We know that the formation of the axis relies on \u201ccrosstalk\u201d between cells, which involves long-range signaling molecules (or ligands) and cell-surface receptors on cells that receive the signal.The molecules involved in the formation of the left-right axis during embryogenesis, along with their functions, are conserved among vertebrates. They include members of the Transforming Growth Factor beta (TGF-\u00df) family\u2014such as the agonists (or ligands) Nodal, Vg1/GDF, and activin, and the antagonist (a molecule that interferes with agonist/ligands) Lefty\u2014on the signaling side and members of the EGF-CFC family\u2014such as the activin receptor and its coreceptors\u2014on the receiving side. The EGF-CFC proteins play important roles in early vertebrate embryogenesis; mutations in these genes in the zebrafish (and mouse) result in a range of developmental defects, including problems in left-right axis specification. While ligand-stimulation of the activin receptor by Nodal and Vg1/GDF requires the EGF-CFC coreceptors, activin can activate the activin pathway without a coreceptor. Lefty\u2014being an antagonist\u2014can block activation of the activin receptor, though it is not clear how.Through genetic and biochemical studies in zebrafish and frog embryos, Simon Cheng, Alex Schier, and colleagues have now clarified a piece of this very complex signaling puzzle by demonstrating that Lefty inhibits a subset of TGF-\u00df signals\u2014Nodal and Vg1/GDF but not activin\u2014by blocking EGF-CFC coreceptors. They went on to show that a short, specific region of the signal molecules\u2014accounting for less than 4% of the entire protein\u2014determines whether the signals activate the activin receptor in an EGF-CFC coreceptor-dependent or -independent fashion and therefore governs susceptibility to Lefty. These findings suggest that subtle sequence differences between related signals can dramatically influence their function.Gene families are thought to arise from gene duplications, and the studies described here illustrate how members of the same gene families can gain diverse roles by specific interactions with coreceptors and antagonists. Additional studies will be necessary to reveal the structural basis for the observed diversity."}
+{"text": "Esophageal dilatation is the most widely used treatment option for the management of esophageal strictures. Complications include bleeding, brain abscess, esophageal perforation and bacteremia. Nasopharyngeal gangrenous abscess after the esophageal dilatation is very rare. Endonasal endoscopic surgery was performed to treat the lesion and a successful result was obtained.Escherichia coli. Symptoms improved after the operation and treatment with antibiotics.A 59-year-old woman with a previous history of dilatation for esophageal stricture was admitted with a low-grade fever, headache, neck pain and cranial nerve abnormalities including sixth nerve palsy. Imaging studies aroused suspicion of necrotic retropharyngeal tumor with clivus, condylar process and cavernous sinus invasion. Biopsy with a pharyngosope was performed by an ENT doctor. The pathology showed acute necrotic inflammation, tissue granulation and bacteria colonies. Navigation with endonasal endoscopic surgery was chosen to treat the skull base and nasopharyngeal abscess. Bacterial culture showed A nasopharyngeal gangrenous abscess with extension to the skull base in the case of esophageal reconstruction after esophageal dilatation is extremely rare. Physicians dealing with esophageal stricture should keep in mind that a nasopharyngeal abscess is a potential complication of esophageal dilatation. Escherichia coli after esophageal reconstruction with esophageal dilatation. The patient was successfully treated by a minimally invasive procedure with endonasal endoscopic approach to treat the abscess. The clinical course and the mechanisms of the abscess formation are discussed.The most common procedure used for the esophageal stricture is esophageal dilatation which has dose associated risks. Esophageal perforation is the worst complication. The perforation rate is reported to range between 0.1 and 0.4%. The rateA 59-year-old woman had a history of esophageal benign tumor after esophagectomy and reconstruction with gastric tube, 4 years ago. She had past history of neck pain radiating to the left shoulder and occipital region. Severe neck pain limited her range of motion. She had been admitted to a hospital with the suspicion of cervical disk herniation with cervical radiculopathy.Two months before admission, she complained of dysphagia, and esophageal dilatation was performed due to the stricture. One month later, she developed diplopia, slurred speech and dysphasia. Low-grade fever and whitish rhinorrhea were also noticed. On neurological examination, the patient was alert and oriented. She had severe neck pain, left sixth nerve palsy, mildly slurred speech and dysphagia. Other cranial nerves, including III, IV, V, had no defect. Muscle strength was normal in upper extremities. The deep-tendon reflexes were normal bilaterally. Sensory examination showed nothing unusual. Laboratory studies were within the normal range. A plain chest film showed a supradiaphragmatic gastric bulb. C-reactive protein levels were elevated to 3.6mg/dL . Magnetic resonance imaging (MRI) of sellar region disclosed a thickening of the nasopharyngeal wall with irregular contrast enhancement. Ill-defined enhancement was seen over the skull base including the left cavernous sinus, left parasellar region, clivus, bilateral petrosal apex, condylar process of the occipital bone and C1 vertebra . NasophaE.coli. Two weeks of Flomoxef therapy was given and the patient was discharged 22 days after operation and she presented with only mild left sixth nerve palsy. Two months later, MRI scan with DWI sequences showed residual abscesses in the left cavernous sinus and parasellar region [We took an endonasal endoscopic approach to treat the suprasellar and nasopharyngeal lesion. Gray brownish necrotic tissue of soft to firm nature was removed . Patholor region .Streptococcus viridans, probably originating as a contaminant from the oropharynx.[E. coli, which is different from previous reports.Esophageal dilatation is the most widely used option for the treatment of esophageal strictures. It is associated with bacteremia in 22 -72% of the procedures2 whereas,opharynx. However,opharynx. The caseStreptococcus pyogenes followed by Staphylococcus aureus.[E. coli infections have been reported only in a few cases.[E. coli infection in this case included esophageal stricture related to the supradiaphragmatic gastric bulb, esophageal dilatation complicated with retropharyngeal abscess and contiguous spread of inflammation to the cavernous sinus, parasellar region and clivus. Abducens palsy has been reported as a common complication of the clivus and paraclival pathology.[Retropharyngeal abscesses are uncommon but are potentially lethal. Abscesses in adults are usually secondary to the trauma of upper aerodigestive tract caused by foreign bodies or iatrogenic instrumentation such as dental procedures, feeding tube insertion, oral tracheal intubation, esophagoscopy, etc. The most commonly isolated pathogen is s aureus. In literew cases. None of athology.A minimally invasive endonasal endoscopic surgery was commonly used for approached the abscesses over the clivus, sellar, parasellar region and cavernous sinus. The major advantage of this operation is that it provides the most direct anatomical route to the lesion without transversing the major neurovascular structures. A stereoA nasopharyngeal gangrenous abscess in the case of esophageal reconstruction with extension to the skull base following esophageal dilatation is extremely rare. Tissue sampling is important to differentiate an infectious disease from malignancy. In endonasal endoscopic surgery, the endoscope allows us an extended view over the skull base around the sellar region and onto the surrounding structures. Physicia"}
+{"text": "Controlling and limiting the expression of short hairpin RNA (shRNA) by using constitutive or tissue-specific polymerase II (pol II) expression can be a promising strategy to avoid RNAi toxicity. However, to date detailed studies on requirements for effective pol II shRNA expression and processing are not available. We investigated the optimal structural configuration of shRNA molecules, namely: hairpin location, stem length and termination signal required for effective pol II expression and compared it with an alternative strategy of avoiding toxicity by using artificial microRNA (miRNA) scaffolds.Highly effective shRNAs targeting luciferase (shLuc) or Apolipoprotein B100 (shApoB1 and shApoB2) were placed under the control of the pol II CMV promoter and expressed at +5 or +6 nucleotides (nt) with reference to the transcription start site (TSS). Different transcription termination signals (TTS), namely minimal polyadenylation (pA), poly T (T5) and U1 were also used. All pol II- expressed shRNA variants induced mild inhibition of Luciferase reporters carrying specific targets and none of them showed comparable efficacy to their polymerase III-expressed H1-shRNA controls, regardless of hairpin position and termination signal used. Extending hairpin stem length from 20 basepairs (bp) to 21, 25 or 29 bp yielded only slight improvement in the overall efficacy. When shLuc, shApoB1 and shApoB2 were placed in an artificial miRNA scaffold, two out of three were as potent as the H1-shRNA controls. Quantification of small interfering RNA (siRNA) molecules showed that the artificial miRNA constructs expressed less molecules than H1-shRNAs and that CMV-shRNA expressed the lowest amount of siRNA molecules suggesting that RNAi processing in this case is least effective. Furthermore, CMV-miApoB1 and CMV-miApoB2 were as effective as the corresponding H1-shApoB1 and H1-shApoB2 in inhibiting endogenous ApoB mRNA.Our results demonstrate that artificial miRNA have a better efficacy profile than shRNA expressed either from H1 or CMV promoter and will be used in the future for RNAi therapeutic development. RNA interference (RNAi) is an evolutionary conserved mechanism for regulating gene expression. It plays an important role in defense against viruses but also in development and in normal functioning of the cell ,2. The nBy introducing molecules that constitute substrates for the endogenous RNAi pathway disease-related mRNA and thus protein translation can be inhibited. RNAi in cells can be induced in different ways. Generally this is achieved by delivery of 20\u201325\u2009bp-long small interfering RNAs (siRNAs)  which seTo date, shRNA expression from pol III promoters is the most common way to induce RNAi in cells, which provides potent and stable target inhibition. Several pol III promoters are being used for expression of shRNAs, such as H1 or U6, and transcription initiation and termination sites together with the structural requirements for effective expression of the hairpins are well described ,8. Howevin vivo toxicity related to high shRNA expression levels from a pol III promoter was abolished when the shRNA was expressed from the liver-specific pol II hAAT promoter and U1 termination signal was used [This toxicity problem may be circumvented by using weaker pol II promoter to express shRNAs or by embedding siRNA into artificial miRNA scaffolds. The CMV promoter has been the first pol II promoter shown to express active shRNA and initial requirements for this approach have been established: the shRNA has to be juxtaposed to the transcription start site (TSS) within 6 nt) and followed by the minimal polyadenylation signal (pA) . When thwas used . Incorpo nt and fwas used .In the current study we focused on the structural requirements to determine the optimal configuration for pol II shRNA expression and compared it to the pol II expression of artificial miRNA. To date, no detailed studies on the hairpin positions, transcription termination signal (TTS), and shRNA stem length requirements for efficient pol II shRNA expression have been performed. Varying positions of the hairpin relative to the TSS +5 or +6, stem length of 20, 21, 25, 29\u2009bp and different TTS  have been tested for a shRNA targeting luciferase (shLuc) and expressed from the pol II CMV promoter. shLuc at +6 location relative to TSS, with stem length of 20 nt with either pA or U1 termination signal was found to yield optimal results. However, the average luciferase inhibition of 40% was still lower than the control pol III H1-shRNA, which inhibited luciferase up to 90%. Additionally, two shRNAs targeting ApoB (shApoB1 and shApoB2) were found inactive when expressed from CMV using the same configuration. In contrast, when the three shRNAs were incorporated in an artificial miRNA scaffold, two were equally active to their respective H1-shRNA controls while expressing much less siRNA molecules. This indicates that using artificial miRNA scaffolds has a greater therapeutic potential than direct expression of shRNAs from pol II promoters.Previously, we have identified a highly effective siRNA potent in knocking down luciferase when expressed as shRNA from the pol III H1 promoter (shLuc) . To miniThe length of the stem in the shRNA has been found to be important for Dicer processing resulting in enhancement of the silencing potential of a given hairpin . Dicer rin vitro and in vivo the expression of ApoB that is the structural protein of low density lipoprotein cholesterol (LDL-C) [Previously, H1-shApoB1 and H1-shApoB2 have been used to inhibit  (LDL-C)  . In conclusion, we and others have shown that pol II expression of artificial miRNA scaffolds is a more robust and promising approach than pol II shRNA expression, probably due to the specific structural characteristics of the miRNA.An alternative to optimizing pol II expression of shRNA is to use pre-miRNA scaffolds and replace the mature miRNA sequence with siRNA targeting a gene of interest. Cellular miRNAs are naturally expressed from pol II promoters ,27. Therof shRNA . Similarof shRNA . In a prAn additional advantage of the use of artificial miRNAs is that several precursors can be expressed as clusters from longer transcripts, allowing simultaneous targeting of multiple genes . This feet al., manuscript submitted). The siRNAs originating from the H1-shRNA were more heterogeneous in cleavage sites and length compared to the products originating from the CMV-miRNA scaffold supporting the notion that Dicer cleavage is less precise than the combination of Drosha and Dicer. Unfortunately, NGS data from pol II-expressed shRNA, which would allow verifying the processed siRNA variants and their abundance, are still not available.Quantification of the siRNA molecules expressed from the H1-shRNA, CMV-shRNA or CMV-miRNA constructs was highly specific and revealed differences in the amount of processed siRNA molecules per cell. As expected, highest siRNA amounts were detected when the shRNAs were expressed from the strong H1 promoter. However, when the same shRNAs were expressed from the CMV promoter, 12- to 125-fold less siRNA molecules were detected indicating that either the shRNA was not efficiently transcribed from the CMV promoter or that there was impaired siRNA processing by the RNAi machinery. Surprisingly, the amount of processed siRNA from the CMV-miRNA constructs did not correlate with efficacy. For example, CMV-miLuc yielded 1591 siLuc molecules per cell and was completely ineffective in target knockdown while CMV-miApoB2 yielded only 56 siApoB2 molecules per cell and was highly efficient. A possible explanation for this discrepancy is that the small RNA TaqMan assay detects only one variant of the guide strand and although identical predicted siRNA sequences were incorporated in the CMV-shRNA and CMV-miRNA scaffolds, they can still be processed differently. Indeed, our newest data based on Next Generation Sequencing (NGS) of small RNAs from cells transfected with H1-shApoB2 and CMV-miApoB2 indicates differential processing and different mature sequences from the two scaffolds . The vector contains a stretch of five thymidines (T5), which acts as the transcription termination signal. Panel of shRNAs were designed to act as positive controls: one shRNAs was designed that target Firefly luciferase (H1-shLuc), two shRNAs were made that target different regions of human ApoB gene (H1-shApoB1 and H1-shApoB2). Additionally two negative controls shRNA were deigned: one that do not target any gene in human and mouse genome (H1-shScr) and one that target the GFP (H1-shGFP). All the sequences of oligonucleotides used in this study are listed in Additional file CMV-shRNA with different termination signals were designed as followed. First different TTS  were annealed from complementary oligonucleotides and ligated into the AccI and SphI sites of pVD23 which includes the CMV promoter. Next, plasmids were digested with SacI and NheI and complementary oligonucleotides containing shRNA targeting GFP (shGFP) or Luciferase (shLuc) were inserted. shLuc oligonucleotides were designed to allow the hairpin transcription to start at +5 or +6 nucleotides after the TSS, which was considered as +1. The final constructs were CMV\u2009+\u20095shLuc-T5, CMV\u2009+\u20096shLuc-T5, CMV\u2009+\u20095shLuc-pA and CMV\u2009+\u20096shLuc-pA. For U1 termination signal only CMV\u2009+\u20096shLuc-U1 was created. As negative controls CMV-shGFP-T5, CMV-shGFP-pA and CMV-shGFP-U1 were made. GFP oligonucleotides were designed that hairpin started at +6 from the TSS.CMV-shRNAs with different stem length of the hairpin were designed as followed. CMV-shLuc20, CMV-shLuc21, CMV-shLuc25 or CMV-shLuc29 were made by annealing complementary oligonucleotides that create the stem-loop structure of 20, 21, 25 and 29\u2009bp respectively and ligating them into the BamHI and XhoI sites of the pSilencer 4.1 CMV vector . Constructs contained either the pA or the U1 termination signal. To create the CMV-shApoB1 and CMV-siApoB2, siRNA sequences from shApoB1 and shApoB2 were cloned by annealing complementary oligonucleotides and ligating them into the BamHI and XhoI sites of the pSilencer 4.1 CMV vector with pA termination signal. As the negative control, pSilencer 4.1 CMV control  plasmid was used, named CMV-shScr.To create the artificial miRNA expressing vectors, siRNA sequences from shLuc, shApoB1 and shApoB2 were cloned into the pri-mir-155 backbone of pcDNA6.2-GW/EmGFP-miR  by annealing complementary oligonucleotides and ligation into BamHI- and XhoI-digested pcDNA6.2 plasmid and named respectively CMV-miLuc, CMV-miApoB1and CMV-miApoB2. As the negative control pcDNA6.2-GW/EmGFP-miR-neg control  was used, named CMV-miScr. The Mfold program was used to determine the secondary structure of all RNA transcripts .Reporter constructs that were used for luciferase knockdown are Firefly luciferase pGL4  and pRL Renilla  under control of the CMV promoter. Luc-ApoB1 reporter, containing in its 3\u2019 UTR a fragment of 239 nucleotides of human ApoB  sequence was made, by cloning of the ApoB sequence, that was PCR amplified with primers pr565f and pr566r in the XhoI and NheI sites of pGL4 plasmid. For The Luc-ApoB2 containing in the 3\u2019UTR of siCheck2 vector  1851 nucleotides of the last exon from the human ApoB has been described previously.The human embryonic kidney (HEK)293\u2009T and mouse hepatoma Hepa1-6 cell lines were maintained in Dulbecco\u2019s modified Eagle\u2019s medium  containing 10% fetal calf serum, 100 U/ml penicillin and 100 U/ml streptomycin, at 37\u00b0C and 5% CO2. For luciferase assays, endogenous ApoB knockdown assays, and siRNA expression analysis, cells were seeded in 96- or 24-well plates at a density of 3*104 or 1.2*105 cells per well, respectively, in DMEM one day prior transfection. Transfections were performed with Lipofectamine 2000 reagent  according to the manufacturer\u2019s instructions.Cells were co-transfected with 100\u2009ng shRNA or miRNA expressing plasmid and respectively 2,5\u2009ng Firefly luciferase or Luc-ApoB1 and 0,5\u2009ng Renilla or 50\u2009ng Luc-ApoB2 reporter. Transfected cells were assayed at 48\u2009hr post-transfection in 20\u2009\u03bcl 1x passive lysis buffer  Firefly and Renilla were measured luciferase activities with the Dual-Luciferase Reporter Assay System . For luciferase knockdown calculations, H1-shLuc was used as a positive control. CMV-shScr, CMV-shGFP or CMV-miScr served as negative controls and were set at 100%. For luciferase- and ApoB1- and ApoB1 targeting constructs Firefly luciferase expression was normalized to Renilla luciferase expression. For ApoB2-targeting constructs Renilla luciferase expression was normalized to Firefly luciferase expression since the siCheck2 plasmid was used.in vitro, Hepa1-6 cells were transfected with 1\u2009\u03bcg shApoB1, shApoB2, miApoB1 or miApoB2 and total RNA was isolated from cells 48\u2009hr post-transfection using the Nucleospin kit . Genomic DNA (gDNA) was removed by DNase treatment using TURBO DNase . First strand cDNA was reverse transcribed using random hexamer primers with the Dynamo kit . Real time PCR amplification was performed with ApoB- and beta actin-specific primers  using 10\u2009ng RNA isolated from cells transfected with shRNA or miRNA expression plasmids and 3\u2009\u03bcl custom-made specific RT-stem-loop primers  according to the manufacturer\u2019s instructions. The TaqMan assay was done in 20\u2009\u03bcl using 1,33\u2009\u03bcl cDNA, 1-\u03bcl custom-made siRNA-specific primer with FAM-labeled fluorogenic probe (Applied Biosystems) and 10\u2009\u03bcl TaqMan 2\u00d7 Universal PCR Master Mix (Applied Biosystems). For determining the assay specificity, 10\u2009ng of the indicated oligo or expression plasmid was added to the TaqMan reaction. Amplification of the beta actin gene was used as a RNA quality and loading control. siRNA copy number per cell was calculated based on the amplification plot of a dilution series of synthetic siRNA standards . Tenfold dilutions (100\u2009pg \u2013 1\u2009fg) of the guide strand of the synthetic siRNA oligo were used to create a standard line. The copy number of each dilution was calculated according to the formula 1\u2009mol\u2009=\u20096.02\u2009\u00d7\u200910per cell .PM, FB, HP, SvD and PK are employees of uniQure biopharma b.v. AK and TR are former employees of uniQure biopharma b.v. The authors declare that they have no competing interests.PM carried out the experiments, designed the study and prepared the manuscript; AK, FB, HP, SvD and TR participated in the study design and drafting of the manuscript; PK designed and interpreted the research and participated in preparing of the manuscript. All authors read and approved the final manuscript.Table S1. Oligonucleotides used in this study.Click here for fileTable S2. Determination of siApoB1 RT primers and TaqMan probe specificity on siApoB1, shApoB1 hairpin precursor, shApoB1- and miApoB1-expression plasmid and siApoB2.Click here for fileTable S3. Determination of siApoB2 RT primers and TaqMan probe specificity on siApoB2, shApoB2 hairpin precursor, shApoB2- and miApoB2-expression plasmid and siApoB1.Click here for file"}
+{"text": "Chronic musculoskeletal (MS) pain is common in patients with chronic kidney disease (CKD) undergoing haemodialysis. However, epidemiological data for chronic MS pain and factors associated with chronic MS pain in patients with early- or late-stage CKD who are not undergoing dialysis are limited.A cross-sectional study to evaluate the prevalence of chronic MS pain and factors associated with chronic MS pain in patients with early- and late-stage CKD who were not undergoing dialysis, was conducted. In addition, the distribution of pain severity among patients with different stages of CKD was evaluated.Of the 456 CKD patients studied, 53.3% (n\u2009=\u2009243/456) had chronic MS pain. Chronic MS pain was independently and significantly associated with hyperuricemia as co-morbidity, as well as with the calcium\u2009\u00d7\u2009phosphate product levels. In CKD patients with hyperuricemia, chronic MS pain showed a negative, independent significant association with diabetes mellitus as a co-morbidity . However, in the CKD patients without hyperuricemia as a co-morbidity, chronic MS pain showed an independent significant association with the calcium\u2009\u00d7\u2009phosphate product levels . Furthermore, stage-5 CKD patients seemed to experience more severe chronic MS pain than patients with other stages of CKD.Chronic MS pain is common in CKD patients. Chronic MS pain was independently and significantly associated with hyperuricemia as co-morbidity, and with the calcium\u2009\u00d7\u2009phosphate product levels in early- and late-stage CKD patients who were not on dialysis. The worldwide prevalence of chronic kidney disease (CKD) continues to increase . CKD patCKD patients have a poor quality of life . This maIt has been reported that 82% of CKD patients undergoing dialysis had chronic pain, . ChronicDespite the understanding of prevalence and risk factors associated with chronic MS pain in CKD patients undergoing dialysis, the epidemiologic data for factors associated with chronic MS pain in early- and late-stage CKD patients who are not on dialysis, are limited. In addition, information about the prevalence of non-steroidal anti-inflammatory drugs (NSAID) or Chinese herbs used for pain relief in CKD patients is also limited. The aims of this study were to determine the prevalence of chronic MS pain in CKD patients and to identify the factors associated with chronic MS pain.2 [determined by the CKD Epidemiology Collaboration (CKD-EPI) creatinine equation], in two separate measurements within an interval of 3\u00a0months  and waist circumference), history of smoking, alcohol and betel nut consumption, current use of NSAIDs or Chinese herbal medicines, and the presence of diabetes, hypertension, coronary artery disease (CAD), peripheral arterial disease (PAD), congestive heart failure (CHF), stroke, hyperuricemia, systemic lupus erythematosus (SLE), urolithiasis, polycystic kidney disease (PKD), rheumatoid arthritis (RA), spondylarthropathies, spine osteoarthritis (OA), and back pain. Current use of diuretics or allopurinol was also collected. Baseline haematological and biochemical data of these patients were also collected. Smoking was defined as the consumption of more than 10 cigarettes per day for at least one year. Alcohol use was defined as daily consumption of more than 80\u00a0mg of alcohol/day for more than one year. Current use of NSAIDs was defined by self-reported use of ibuprofen, naproxen, sulindac, piroxicam, indomethacin, tolmetin, or diclofenac (with brand names and combination formulas identified) daily or nearly every day, for the last 30\u00a0days. Current use of Chinese herbs was defined by self-reported use of any Chinese herb daily or nearly every day, for the last 30\u00a0days. Diabetes mellitus was defined as a fasting glucose level \u2265126\u00a0mg/dL or use of any hypoglycaemic medication. Hypertension was considered to be present if the patient was on antihypertensive medicine, or if blood pressure was >140/90\u00a0mm Hg. The presence of CAD was confirmed by coronary angiography, and CHF was diagnosed on the basis of the Framingham criteria for heart failure . PAD wasFull laboratory profiles were obtained. The laboratory parameters included levels of blood urea nitrogen, serum creatinine (Scr), haemoglobin, albumin, calcium (Ca), phosphate (P), intact parathyroid hormone (iPTH), total cholesterol, triglycerides, uric acid, and high-sensitivity C-reactive protein (hs-CRP). Serum creatinine levels were assessed by spectrophotometric analysis using a modified kinetic Jaffe reaction.p\u2009<\u20090.05 in the univariate analysis were selected. All analyses were performed by using the commercially available statistics software, SPSS version 15.0 for Windows.Descriptive statistics were expressed as means with standard deviation or percentage frequency, as appropriate. All variables were tested for normal distribution by a Kolmogorov-Smirnov test. The Student\u2019s t-test or Mann\u2013Whitney U test was applied to compare means of continuous variables. Categorical data were tested using the chi-square test. Binary logistic regression was appropriately used to test the factors associated with chronic MS pain. To identify independent associations, stepwise multivariate regression analysis was used, and variables including age, gender, and Mean age of the patients was 63.3\u2009\u00b1\u200914.1\u00a0years and 42.5% (n\u2009=\u2009194/456) of patients were male. The NSAID and Chinese herb use by CKD patients was around 7.8% (n\u2009=\u200940/456), and 9.6% (n\u2009=\u200944/456), respectively. Approximately 30.7% (n\u2009=\u2009140/456) of patients had DM as a co-morbidity (Table\u00a0CKD patients were divided into three groups: early CKD (CKD stage 1\u20132), CKD stage 3\u20134, and CKD stage 5, in order to compare the percentage of patients with chronic MS pain between groups. It was found that similar prevalence of chronic MS pain was noted in early CKD, CKD stage 3\u20134, and CKD stage 5   of chronic MS pain, moderate pain about 28.4% (n\u2009=\u200969/243), and mild pain only accounted for 15.6% (n\u2009=\u200938/243) of chronic MS pain. Mild chronic MS pain was noted in 17.8% (n\u2009=\u200921/118) of CKD patients with stage 1\u20132 disease, 13.3% (n\u2009=\u200915/113) of CKD patients with stage 3\u20134 disease, and 16.7% (n\u2009=\u20092/12) of CKD patients with stage 5 disease (p\u2009=\u20090.594). Moderate chronic MS pain was noted in 28% (n\u2009=\u200933/118) of CKD patients with stage 1\u20132 disease, 31.9% (n\u2009=\u200936/113) of CKD patients with stage 3\u20134 disease, and 0% (n\u2009=\u20090/12) of CKD patients with stage 5 disease (p\u2009<\u20090.001). Severe chronic MS pain was found in 54.2% (n\u2009=\u200964/118) of CKD patients with stage 1\u20132 disease, 54.9% (n\u2009=\u200962/113) of CKD patients with stage 3\u20134 disease, and 83.3% (n\u2009=\u200910/12) of CKD patients with stage 5 disease (p\u2009<\u20090.001).2/mL2, p\u2009=\u20090.039) were also linked with the presence of chronic MS pain. Chronic MS pain was not associated with age, smoking, alcohol/betel nut consumption, or co-morbidities such as hypertension, CAD, PAD, CHF, stroke, SLE, urolithiasis, PKD, RA, spondylarthropathies, spine OA, and diuretics use. In particular, there was no significant difference between current NSAID and Chinese herb use in patients with and without chronic MS pain . The presence of chronic MS pain was also not associated with renal function , albumin, Ca, P, iPTH, cholesterol, triglycerides, uric acid, or hs-CRP.Compared with patients who experienced no chronic MS pain, those who reported chronic MS pain were more likely to be male, have a higher BMI and waist circumference, higher systolic blood pressure, and were less likely to have DM as a co-morbidity. In particular, chronic MS pain in CKD was related to hyperuricemia and back pain co-morbidities. One hundred seventy two patients  who had hyperuricemia had chronic MS pain, but only 71 patients  without hyperuricemia had chronic MS pain (p\u2009<\u20090.001)  in our CKD study patients with chronic MS pain as compared to the report of Pham et al. (23.2%) . This maThe present study revealed that 53.3% patients had chronic MS pain, with a mean eGFR around 65\u00a0ml/min. However, Cohen et al.  reportedThe results of the present study indicate a correlation between the co-morbidity of hyperuricemia, the calcium\u2009\u00d7\u2009phosphate product levels, and the co-morbidity of DM with chronic MS pain in CKD patients. This study raises the importance of uric acid control and gout prevention to decrease the possibility of chronic MS pain, thereby improving the quality of life in CKD patients. In addition, the study found that the calcium\u2009\u00d7\u2009phosphate product levels were an important factor for chronic MS pain, especially in CKD patients without hyperuricemia. Results of this study demonstrate that the calcium\u2009\u00d7\u2009phosphate product levels are an important laboratory parameter to consider while treating CKD patients with chronic MS pain and without hyperuricemia. Additionally, it was demonstrated that NSAID or Chinese herb use was not very common in CKD patients. Low prevalence of NSAID and Chinese herb use (7.8% and 9.6%) may indicate either good patient education by healthcare professionals, or an underestimation or inaccurate reporting of NSAID and Chinese herb use by patients.This study had several limitations. The information regarding pain relief agents was lacking. Furthermore, the colchicine use for gout relief in study patients was not collected. In addition, quality of life was not evaluated in this study. However, we believe that the results provide a solid basis for studies that will further explore these relationships. The ultimate goal should be a better understanding and treatment of chronic MS pain in CKD patients.In summary, the present study demonstrated that chronic MS pain was independently and significantly associated with hyperuricemia and the calcium\u2009\u00d7\u2009phosphate product levels. In addition, in CKD patients with hyperuricemia, chronic MS pain was negatively, independently, and significantly associated with DM. However, in CKD patients without the co-morbidity of hyperuricemia, chronic MS pain was independently and significantly associated with the calcium\u2009\u00d7\u2009phosphate product levels.The authors declare no financial competing or other competing interest.H-J H participated in data collection, statistical analysis and manuscript preparation; C-H Y, K-H H, and C-C L participated in statistical analysis; I-W W, M-J H, C-Y S, C-C C, Y-C C helped with data collection; M-F H, C-Y C, C-Y H, and C-J T participated in data interpretation; M-S W participated in study design, coordination, and helped to draft the manuscript. All authors read and approved the final manuscript.Heng-Jung Hsu and Chiung-Hui Yen are Co-first authors.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2369/15/6/prepub"}
+{"text": "There are over a half a million copies of L1 retroelements in the human genome which are responsible for as much as 0.5% of new human genetic diseases. Most new L1 inserts arise from young source elements that are polymorphic in the human genome. Highly active polymorphic \u201chot\u201d L1 source elements have been shown to be capable of extremely high levels of mobilization and result in numerous instances of disease. Additionally, hot polymorphic L1s have been described to be highly active within numerous cancer genomes. These hot L1s result in mutagenesis by insertion of new L1 copies elsewhere in the genome, but also have been shown to generate additional full length L1 insertions which are also hot and able to further retrotranspose. Through this mechanism, hot L1s may amplify within a tumor and result in a continued cycle of mutagenesis.We have developed a method to detect full-length, polymorphic L1 elements using a targeted next generation sequencing approach, Sequencing Identification and Mapping of Primed L1 Elements (SIMPLE). SIMPLE has 94% sensitivity and detects nearly all full-length L1 elements in a genome. SIMPLE will allow researchers to identify hot mutagenic full-length L1s as potential drivers of genome instability. Using SIMPLE we find that the typical individual has approximately 100 non-reference, polymorphic L1 elements in their genome. These elements are at relatively low population frequencies relative to previously identified polymorphic L1 elements and demonstrate the tremendous diversity in potentially active L1 elements in the human population.The online version of this article (doi:10.1186/s12864-015-1374-y) contains supplementary material, which is available to authorized users. Mobile elements have been extremely successful at colonizing human genomes. As much as two thirds of human genomic content is either made up of mobile elements or mobile element relics ,2. The tL1 elements have been amplifying in mammalian genomes since before the divergence of placental and marsupial mammals 170 million years ago . Since bActive full length L1 is a ~6\u00a0kb long element that codes for two open reading frames (ORFs): ORF1 which encodes a protein with nucleic acid chaperone and RNA binding properties, and ORF2 which encodes a protein with endonuclease (EN) and reverse transcriptase (RT) activities ,16. L1 mAn estimated 10-30% of new L1 insertions are full length with the potential for further retrotransposition . NegativStudies that have looked at full-length L1 elements in the reference build of the human genome have indicated that the bulk of retrotransposition occurs from a handful of hot L1 elements . AdditioIn this report, we describe a NGS-based method to identify full length L1 elements in human genomes that we call Sequencing Identification and Mapping of Primed L1 Elements (SIMPLE). Using SIMPLE we have identified 228 polymorphic L1 elements in seven independent individuals. SIMPLE has 94% sensitivity and is robustly able to identify nearly all full length L1 elements in a human genome, making it possible to identify hot L1s capable of causing high degrees of insertional mutagenesis.Various anchored PCR strategies have been effectively employed to identify mobile element insertions in a whole-genome setting. They all share the use of one primer from within the known sequence of the element, and use different strategies to target a second primer outside of the element. However, there are several significant drawbacks to most of those methods. Those that ligate a linker to a restriction site located randomly outside of the mobile element insertion only create one amplified fragment size from each element. Thus, it is impossible to tell whether duplicate sequences are independent ligations or PCR duplicates. A similar argument can be made for the use of an arbitrary primer to prime outside the mobile element. Furthermore, restriction sites located at different lengths away from the element will amplify with differential efficiencies, as will different arbitrary primers. Thus, these methods require pooling of multiple experiments using different restriction enzymes or primers to come close to saturating the potential insertions. In addition, even those protocols that utilize randomly sheared DNA to apply anchors still use a major ligation step in their preparation. These ligation steps, such as those used in the preparation of Illumina libraries, lead to low levels of chimeric fragment ligation between genomic fragments that can confound data analysis .Sequencing Identification and Mapping of Primed L1 Elements (SIMPLE) is a unique L1-detection method based on the principles of random shearing combined with t-linker ligation-mediated PCR that takes advantage of the massive parallelization offered by high throughput NGS technology . A primeHomo sapiens, and thus represent L1 elements universally present in all human genomes at a diploid level. We employed SIMPLE to determine how many known, fixed full-length L1 elements on three randomly selected chromosomes we could detect. We limited our initial analysis to three chromosomes to allow a more exhaustive manual analysis of any variants. Of the 543 fixed full-length L1 elements on three randomly selected chromosomes, SIMPLE successfully detected the vast majority with an average read depth of ten independent (having a different linker location) reads per element. Of these, 511/543 (94%) were detected by at least one SIMPLE read, with 502/543 (92%) detected by more than three independent SIMPLE events (reads)  we assessed the detection rate of previously described polymorphic full-length L1 elements on the same three chromosomes ,25-27. WTo determine the extent of human full length L1 polymorphism between unrelated individuals, we applied SIMPLE to seven individual genomic DNA samples from non-related Caucasian individuals to assess the degree of full-length L1 polymorphism between these individuals. For ease of analysis, SIMPLE reads mapping to known full-length L1 elements were discarded, and only those reads that did not represent reference elements were taken into account for analysis.Using SIMPLE, we detected 228 non-reference full-length L1 elements in the genomes of seven unrelated individuals. The range of non-reference, full-length L1 elements varied from a low of 73 per individual to a high of 134 per individual with an average of 96 non-reference full length L1 elements per individual  SIMPLE-detected L1 polymorphisms by 5\u2019 junction PCR and confirmed the insertions by Sanger sequencing of the 5\u2019 junction  because the vast majority of new L1 insertions are 5\u2019 truncated ,20,36,37de novo somatic L1 insertion events in tumors or other samples. Because DNA fragmentation for SIMPLE occurs by random shearing of genomic DNA by sonication, SIMPLE is able to detect L1 elements that may be located too distally from restriction enzyme cut sites used by other high-throughput sequencing methods [Full length polymorphic L1 elements are responsible for virtually all of the disease-causing retroelement insertions and the bulk of retroelement mobilization observed in tumors. We have developed a robust tool to identify full-length polymorphic L1 elements in individual genomes, which also has potential to be applied to the detection of  methods ,26. Addi methods . Those rUsing SIMPLE, we have determined the typical individual contains about 100 non-reference, full-length L1 elements. Based on previous estimates, we expect 63/100 of these elements to be active, with 54/100 exhibiting particularly high levels of activity . These 1It is interesting to note that although all seven individuals assayed in this study were Caucasian, there is still significant variation between individuals with respect to non-reference, full-length L1 elements. We anticipate, however, that analysis of diverse populations will yield similar results in terms of absolute numbers to those reported here. We would expect an individual from any single population to harbor the average 100 non-reference full length L1 elements with the only difference being in the identity of the particular elements found in that individual. Two individuals from similar backgrounds will share a greater number of non-reference elements than two individuals from diverse backgrounds, but both individuals will have ~100 non-reference full length L1 elements each regardless of their background.SIMPLE represents a useful tool not only for both population and tumor-based studies of L1 insertion polymorphism. The vast majority of L1 retrotransposition is likely to occur due to the approximately 100 polymorphic L1 loci in each individual. Work looking at L1 mobilization in tumors has identified a number of tumor types that appear to support L1 mobilization  as well as others that do not  -27,39,40Fibroblast cell lines GM01631, GM01632, GM05510, GM05568, GM16094, GM15983, and GM08207 were obtained from the Coriell Institute . Cell lines were maintained in EMEM supplemented with non-essential amino acids, sodium pyruvate, and 10% fetal bovine serum. DNA oligonucleotides and duplex linkers were obtained from Integrated DNA Technologies . Oligonucleotide sequences used in this study are presented in Additional file Genomic DNA from fibroblast cell lines was extracted using the DNEasy Blood and Tissue Kit from Qiagen . DNA was sheared to approximate 750-1200\u00a0bp using a Diagenode BioRuptor on High, 30s on/30s off for 12\u00a0minutes. 50\u00a0ng of sheared gDNA was subject to a primer extension reaction using Taq polymerase and a L1 5\u2019 UTR specific primer (L15\u2019UTRP1) which sits ~100\u00a0bp from the start of the L1 element. Phosphorylated duplex T-linkers  were ligated using T4 DNA ligase. First round PCR was performed for 20\u00a0cycles using primers L15\u2019UTRP1 and LinkerP1. One-million fold dilution of PCR I was performed and 1uL of this dilution was subjected to a nested PCR using primers L15\u2019UTRP2 and LinkerP1 for 25\u00a0cycles. PCR products were run on a gel and a gel slice at ~500-700\u00a0bp was extracted using the Qiagen Gel Extraction kit. Following extraction, the final library was amplified using Phusion polymerase  for 12\u00a0cycles as per the Illumina library generation protocol, and gel purified to yield the final 500-700\u00a0bp library.Illumina sequencing libraries were quantified by qPCR and checked for quality by Agilent BioAnalyzer trace at Elim Biopharmaceuticals . 100\u00a0bp paired end Illumina sequencing was performed on an Illumina HiSeq 2000 by Elim Biopharmaceuticals .Reference repetitive element annotation files were downloaded from UCSC genome browser as GTF files and used to annotate the GRCh37 (hg19) reference genome in CLC Genomics Workbench . Two custom GFF annotation files were also generated: 1) representing known polymorphic elements and 2) representing all known full length L1 elements plus 600\u00a0bp of upstream genomic flanking sequence.To identify and annotate 5\u2019 L1 regions of the genome, the first 300 base pairs of the L1.3 consensus sequence was aligned to the human reference genome (CRCh37/hg19) via NCBI BLAST . Initial search parameters were relaxed to increase sensitivity for older, more mutated elements . Blast output was generated in tabular format. Because many of the hits obtained from this search consisted of isolated smaller regions (20-30\u00a0bp) that were independent of any identifiable L1 element, we further filtered for those hits where alignments were made across\u2009>=250\u00a0bp of the 300\u00a0bp query. This greatly improved our specificity while maintaining sensitivity to older, more mutated elements. Tabular format blast results were converted to GFF via Perl scripting, and an additional 600\u00a0bp of flanking sequence was added upstream and downstream to each matching position.Annotation of 3\u2019 L1 ends and flanking sequenced was performed using a similar strategy as above, except that initial identification of 3\u2019 end locations was based on the UCSC Genome Browser hg19 RepeatMasker table track, which allowed for subfamily classification based on all available sequence from the insertion.de novo full length L1 elements.FASTQ sequencing files were analyzed for quality using FastQC. PCR duplicate reads were removed using custom in-house Perl scripting. Sequence read mappings were performed using CLC Genomics Workbench. Single end genomic flank reads were mapped using a two-pronged mapping strategy. First, reads that either mapped to within 600\u00a0bp of known, fixed full length L1 elements or those reads that did not map unambiguously were discarded, as they represented either known elements or unmappable reads. The remaining reads were then mapped uniquely to an annotated GRCh73/hg19 reference genome to identify novel polymorphic or potentially Confirmed novel L1 elements were genotyped to determine the allele frequency of the insert on a DNA panel of 80 diverse individuals  obtained from the Coriell Institute for Medical Research, Camden, NJ. The efficacy of each primer pair was initially assessed through PCR using human DNA cell line HeLa (ATCC CCL-2) and/or chimpanzee DNA cell line Clint (S006006), confirming the size of the predicted PCR products.BLAT was used to locate candidate L1 sequences and flanking DNA these sequences in the hg 19 human reference genome, and add 1,000\u00a0bp of flanking sequence both upstream and downstream. Sequences were then screened through RepeatMasker confirming the absence of candidate L1 inserts from the hg 19 sequence and localizing unique genomic regions for primer design. For the most part, prospective PCR primers flanking each candidate L1 locus were designed using Primer3 software. Once screened through the BLAT genome browser, primers were selected for PCR if they were predicted to amplify a single locus. In addition, a virtual PCR was performed for each locus using the in silico function of BLAT to compute the expected PCR product size, annealing temperature, and further verify that only one locus would be amplified. For candidate L1 loci directly flanked by substantial stretches (more than 1\u00a0kb) of repetitive genomic sequence, primers were designed manually. In such instances, flanking sequences (up to 1\u00a0kb) were screened using the BLAT genome browser and aligned with closely reported matches  in BioEdit (Ibis Biosciences). Point mutations, insertions, and/or deletions specific to the candidate L1 loci were manually positioned toward the 3\u2019 end of the primers. In addition, internal primers, purposed to anneal to the 3\u2019 end of a full length human specific L1 element, were available .PCR assays were performed in four stages. (1) An empty site PCR analysis was initially conducted using HeLa DNA. External primers flanking each candidate L1 locus were used to amplify PCR products matching the predicted empty site band lengths previously generated using BLAT\u2019s in silico PCR feature. This assay was capable of amplifying PCR products no larger than 1,500\u00a0bp. As a result, alleles lacking the L1 insertion were exclusively amplified. Neither alleles possessing an L1 insertion nor unsuitable primer pairs amplified PCR products. Thus it was impossible to distinguish suitable primers pairs flanking L1 inserts that were homozygous present in HeLa DNA from an unsuccessful reaction (failure to amplify an existing empty site). Therefore, given the human specific nature of the predicted L1 inserts, chimpanzee DNA was used in a control PCR analysis examining each primer pair that failed to amplify an empty site during the initial PCR. (2) An internal primer test was conducted to verify the presence of novel L1 inserts and verify that they were full length. These PCR analyses were performed using DNA samples from the individual(s) in which the inserts were originally identified, an external primer flanking the predicted 3\u2019 end of the L1, and a 3\u2019 internal primer . (3) A lStages 1, 2, and 4 were performed in 25\u00a0\u03bcL reactions containing 25\u00a0ng of template DNA, 200 nM of each oligonucleotide primer, 1.5\u00a0mM MgCl2, 10\u00d7 PCR buffer , 0.2\u00a0mM deoxyribonucleotide triphosphates and 1 to 2 U Taq DNA polymerase. PCR reactions were performed as follows: initial denaturation at 94\u00b0C for 60\u00a0seconds, followed by 32\u00a0cycles of denaturation at 94\u00b0C for 30\u00a0seconds, 30\u00a0seconds at primer annealing temperature (determined previously with HeLa DNA), and extension at 72\u00b0C for 30\u00a0seconds. PCR reactions were terminated with a final extension at 72\u00b0C for 2\u00a0minutes. Fractionation of 20\u00a0\u03bcL of each PCR product was performed in a horizontal gel chamber on a 2% agarose gel containing 0.2\u00a0\u03bcg/mL ethidium bromide for 45\u00a0minutes at 200\u00a0V. UV-fluorescence was used to visualize the amplified DNA fragments.Stage 3 was performed using Takara LA-Taq (long amplification) DNA polymerase . These PCR reactions were performed in 25\u00a0\u03bcL reactions containing 25\u00a0ng of template DNA and LA-Taq PCR reagents according to the manufacturer\u2019s suggested protocol: initial denaturation at 94\u00b0C for 1\u00a0minute and 20\u00a0seconds, followed by 32\u00a0cycles of denaturation at 94\u00b0C for 20\u00a0seconds, 20\u00a0seconds at the optimized annealing temperature, and a long extension step at 68\u00b0C for eight minutes and 30\u00a0seconds. These long-amplification reactions were terminated with a final extension at 68\u00b0C for 10\u00a0minutes. PCR products were size fractionated on a 1% agarose gel for 150\u00a0minutes to 180\u00a0minutes at 150\u00a0V. UV-fluorescence was used to visualize the DNA fragments.Library generation for 3\u2019-SIMPLE was performed in the same manner as 5\u2019-SIMPLE with the following modifications. Primer extension used L1HsP1 primer. First round PCR was 22\u00a0cycles rather than 20 and used primers L1HsP1 and LinkerP1. Second round PCR used primers L1HsP2 and LinkerP1. All remaining steps of 3\u2019-SIMPLE library generation were identical to those for 5\u2019-SIMPLE.Raw reads were processed for quality and duplicate removal in the same manner as for 5\u2019-SIMPLE. A similar two-pronged mapping strategy was used for 3\u2019-SIMPLE involving mapping reads to known L1Hs elements and then taking unmapped reads and mapping unambiguously to an annotated GRCh37/hg19 genome."}
+{"text": "Impervious encapsulation around Ahmed glaucoma valve (AGV) results in surgical failure raising intraocular pressure (IOP). Dysregulation of extracellular matrix (ECM) molecules and cellular factors might contribute to increased hydraulic resistance to aqueous drainage. Therefore, we examined these molecules in failed AGV capsular tissue. Immunostaining for ECM molecules  and cellular factors (\u03b1SMA and TGF\u03b2) was performed on excised capsules from failed AGVs and control tenon\u2019s tissue. Staining intensity of ECM molecules was assessed using Image J. Cellular factors were assessed based on positive cell counts. Histopathologically two distinct layers were visible in capsules. The inner layer  showed significant decrease in most ECM molecules compared to outer layer. Furthermore, collagen III (p = 0.004), decorin (p = 0.02), lumican (p = 0.01) and chondroitin sulfate (p = 0.02) was significantly less in inner layer compared to tenon\u2019s tissue. Outer layer labelling however was similar to control tenon\u2019s for most ECM molecules. Significantly increased cellular expression of \u03b1SMA (p = 0.02) and TGF\u03b2 (p = 0.008) was detected within capsular tissue compared to controls. Our results suggest profibrotic activity indicated by increased \u03b1SMA and TGF\u03b2 expression and decreased expression of proteoglycan (decorin and lumican) and glycosaminoglycans (chondroitin sulfate). Additionally, we observed decreased collagen III which might reflect increased myofibroblast contractility when coupled with increased TGF\u03b2 and \u03b1SMA expression. Together these events lead to tissue dysfunction potentially resulting in hydraulic resistance that may affect aqueous flow through the capsular wall. Glaucoma drainage devices are useful in treating refractory glaucoma . CommercThe success of drainage devices surgery depends on the formation and maintenance of a permeable capsule around the episcleral plate, through which the aqueous percolates into surrounding tissues by simple diffusion , 5, 6. TThe tissue related factors that determine the permeability of the capsule have been investigated and although better understood, still remain unclear . Active The hydraulic resistance of interstitium influences many aspects of body fluid physiology including fluid drainage from anterior chamber of the eye. Such resistance is attributed to the nature of extracellular matrix that includes the collagens, proteoglycans and glycosaminoglycans (GAGs) . We hypoAll patients were seen at King Khaled Eye Specialist Hospital, Riyadh, Saudi Arabia. The medical records of patients who underwent revision of Ahmed glaucoma valve implant  for uncontrolled IOP with maximal tolerated medical therapy were reviewed retrospectively to obtain clinical information where available. Failure was defined as intraocular pressure that was above target levels on maximum medical therapy as determined by the treating physician. Inclusion criteria included patients with poorly controlled IOP above target as determined by the treating physician (range 22\u201340 mm Hg) where excision of the capsule was deemed, in the opinion of the physician to be beneficial in controlling IOP. Exclusion criteria included neovascular glaucoma or glaucoma where additional factors may influence the tissue response. The revisions were performed at KKESH between year 1995 and 2010. The Ahmed valve revision in all patients involved excision of the varying amounts of capsule surrounding the implant. Archived paraffin embedded tissue blocks from the capsules excised during the revision were retrieved from pathology archives for light microscopy and immunohistochemistry. Control tenon\u2019s tissue obtained during primary insertion of Ahmed valves was processed and embedded in paraffin for the study. Archived tissue blocks were utilized for the study and data was accessed and analysed anonymously.The study was approved by the Institutional Review Board of King Khaled Eye Specialist Hospital and followed the principles established in the Declaration of Helsinki. The IRB waived the need for consent.Formalin-fixed paraffin embedded (FFPE) tissue from excised capsules and control tenon\u2019s tissue were sectioned at 5 \u03bcm thickness and mounted on coated glass slides. The sections were stained with haematoxylin and eosin for histological evaluation and thickness of the two layers measured. The thickest area of the inner and outer wall was identified and measured in an area that was well demarcated. The measurement was done using Olympus CELLSENS software measurement tools . Indirect immunohistochemistry was performed using a Dako automated stainer . Briefly, deparaffinised slides were treated with antigen retrieval solution  in the Pt link module  processed for staining and the reaction was visualized by using Envision Flex Visualization system . For some samples limited tissue was available during staining, and repeat staining was also not possible in these cases. Hence only those cases were considered where data could be reliably reported and specified accordingly in the results for each antibody. Appropriate positive and negative control tissue were used. Digital images were captured using Olympus BX 53 Microscope . The photographs were taken at a standardized exposure time to evaluate and quantify color intensity using ImageJ-color-deconvolution. The deconvolution method has been used to separate the brown DAB chromogen from hematoxylin counterstain on a microscope slide and the measurement of color intensity of specific stains , 32. BriStatistical Package for Social Studies (SPSS-19) IBM Chicago, USA) was used for statistical analysis. Descriptive statistics were used to report demographic characteristics. The intensity of labelling measured was reported as median, 25% quartile and range. The label intensity between cases and control was compared using non parametric analysis and validated by Kruskal Wallis Test. A two sided \u2018p\u2019- value of less than 0.05 was considered as statistically significant.We studied 14 excised AGV capsules and 8 normal tenon\u2019s tissues as controls. The clinical profile of these subjects is summarized in The intensity of immunostaining for all the examined markers in capsules and controls is detailed in We also observed increased cellular expression of \u03b1SMA (p = 0.02) and TGF\u03b2 (p = 0.008) in the fibroblasts within the capsules compared to the controls . The disFibrosis of the capsule surrounding a glaucoma implant remains the main cause of suboptimal pressure control and failure of drainage procedures . In thisHistologic findings in the capsular tissue were similar to previous reports which described an outer more vascular layer with loosely arranged matrix and an inner relatively avascular layer with dense connective tissue , 34. TheIncreased expression of \u03b1SMA, an indicator of activated myofibroblasts which are key effector cells of fibrosis, was seen in our samples as reported , 12. TheAccompanying increased expression of TGF\u03b2 in our samples was the decrease in ECM molecule decorin. Proteoglycan decorin is a naturally occurring TGF\u03b2 antagonist  that preFurthermore, decrease in decorin and lumican might influence the assembly of collagen in the inner capsular layer. In normal cornea lumican maintains orderly collagen fibril arrangement that is vital for corneal transparency \u201358. In cWe recognize that our study has limitations and further investigations will be needed in this regard. Capsules from functional AGV devices would probably be ideal second comparative controls for this study. However, functional capsules are not commonly excised and were not available for this study. Also, capsules examined in our study were mostly late excisions and expression changes in early stages of shunt failure could not be determined. It has been suggested that thickness and permeability of capsule around the implant is regulated by inflammatory/proliferative and apoptotic processes occurring during wound remodelling of the capsular tissue as a result of exposure to glaucomatous aqueous humor , 6. HoweECM has structural functions and increased expression of ECM molecules could be expected to explain the hydraulic resistance. Paradoxically, we found decreased expression of proteoglycans (decorin and lumican), GAGs (chondroitin sulfate) and collagen III. Together with increased TGF\u03b2 and \u03b1SMA this decrease of ECM molecules might indicate interplay of molecules potentially sustaining a profibrotic environment leading to myofibroblast contractility and tissue dysfunction. Such a role for the ECM as regulators of cell signalling is receiving increasing support , 70 and S1 Table(DOCX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file."}
+{"text": "Carlina vulgaris L. were examined for the chlorogenic acid content, mineral content, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity. Two populations originated from natural nonmetallicolous habitats (NN (populations from Nasi\u0142\u00f3w) and NP (populations from Pi\u0144cz\u00f3w)), and one metallicolous population (MB) was collected from Boles\u0142aw waste heap localized at the place of former open-cast mining of Ag-Pb and Zn-Pb ores dating back to the 13th century and 18th century, respectively. The level of Zn, Pb, Cd, Fe, Ni, and Mn was significantly higher in the root and leaves of MB plants as a result of soil contaminations compared to those of the NN and NP ones. The highest antioxidant potency has been showed by the plants growing in a nonmetallicolous habitat. The flower head extracts obtained from the nonmetallicolous populations also contained the largest amount of chlorogenic acid, whereas the lowest was determined in the roots . These studies provide important information on the influence of a habitat on the quality of herbal materials and the content of the biologically active primary and secondary metabolites.The methanol extracts from three populations of  Free radicals that are constantly generated in the human body cause oxidative stress. The ratio of free radicals may be increased by the factors such as drugs, chlorinated compounds, deficiency of natural antioxidants, tobacco, and pollution. Despite naturally occurring antioxidant systems in the human body, free radicals cause lipid, protein, and DNA oxidation. These damages at the molecular level may influence the etiology of diseases, such as cancer, atherosclerosis, diabetes, neurodegenerative disorders, and aging-related diseases. Some evidence indicates that the diet rich in antioxidants may be protective against abovementioned diseases \u20133.One of the special sources of antioxidants is plant-based natural phenolic compounds. Phenolic compounds, known as nonnutritional ingredients in food, constitute one of the most widely occurring groups of phytochemicals with a wide range of physiological properties. They are components of many herbs, fruits, and vegetables, which are associated with the health benefits after their consumption. A clinical trial and epidemiological studies have established that a dietary intake of fruits is strongly associated with a reduced risk of the civilization diseases. In the human body, they play as antiallergenic, antiatherogenic, anti-inflammatory, antimicrobial, antioxidant, and antithrombotic agents. However, their health benefits also depend on their absorption and metabolism, which in turn are determined by their structure, molecular size and solubility in cell wall structure, location of glycosides in cells, and binding of phenolic compounds within the food matrix .Carlina L. genus (Asteraceae) which comprises over 30 species found in Europe and Asia are rich in antioxidants. Since hundreds of years, some of them have been used in traditional medicine in many countries, for example, in Italy, in Spain, in Hungary, in the Balcan countries, and in Poland. They are used for medicinal and nutritional purposes. They act as diuretic, diaphoretic, stomachic, or antibiotic agents. Extracts are used externally in the treatment of skin diseases. Regarding the chemical composition of Carlina spp., reports are scarce and include mainly pentacyclic triterpenes and essential oil. The Strzemski et al.'s previous studies revealed that the Carlina spp. contain a high amount of triterpenes  [Carlina spp. growing wildly in Poland. As part of a program to search for bioactive constituents from Carlina species, this study was focused on the establishment of phenolic compounds , mineral content, and antioxidant properties of Carlina vulgaris L. Moreover, the differences between the populations from a natural habitat and from a postindustrial area contaminated with heavy metals were investigated.Plants belonging to the ic acid) \u20139. ThereCarlina vulgaris L. (Asteraceae) were collected from three different sites in Poland. Two populations were from Nasi\u0142\u00f3w  and Pi\u0144cz\u00f3w , and one population  originated from a contaminated metalliferous area in Boles\u0142aw. The coordinates of sites and exemplary plant photos are presented in Lebensgeschichte der Gold - und Silberdisteln\u201d [Polish Plants\u201d [Fifty-two specimens of disteln\u201d  and \u201cPol NP, n = , and oneChlorogenic acid standard (\u226595%), 2,2\u2032-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin\u2013Ciocalteau reagent, trolox, aluminum chloride, suprapure nitric acid (65%), and solution of metal standards (1000\u2009ppm) were purchased from Sigma . Methanol, trifluoroacetic acid (TFA), and HPLC-grade acetonitrile were from Merck . Water was deionized and purified by Ultrapure Millipore Direct-Q\u00ae 3UV-R .3\u2009:\u2009H2O  in TOPwave apparatus . Mineralization parameters are given in Table S1 in Supplementary Material available online at https://doi.org/10.1155/2017/1891849. The analysis of metal content was conducted using a high-resolution continuum source atomic absorption spectrometer (HR CS AAS)  in an electrothermal graphite furnace mode for Ag, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, and Pb and using a C2H2/air flame technique for Zn. The validation parameters are summarized in Table S2.Dried and pulverized roots, leaves, and flower heads (0.1000\u2009g) were mineralized using 10\u2009mL of mixture HNOMethanol extracts were prepared according to the previously published procedure . Dried aThe assay was carried out using a Bio-Rad Benchmark Plus microplate spectrometer . Antioxidant capacities of methanol extracts were determined with the use of 2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and expressed as trolox equivalent per gram of air-dry weight (mg TE/g ADW). The total phenolic content (TPC) was established using the Folin\u2013Ciocalteau reagent, and total flavonoid content (TFC) was analyzed based on the reaction with aluminum chloride. TPC and TFC were expressed as equivalent of gallic acid (mg GAE/g ADW) and rutin (mg RUE/g ADW), respectively. All experiments were performed in triplicate.\u03bcm particle size) . The condition of extract separation was based on literature [Chromatographic determination was performed on VWR Hitachi Chromaster 600 chromatograph set coupled with a diode array detector (DAD)  and C18 column Kinetex  was applied to the evaluation of difference between the populations. Differences were determined using Fisher's least significance difference test at the 0.05 probability level. Statistical analysis was carried out using Statistica ver. 12 .Since the environmental conditions may affect significantly the composition and antioxidant properties of a plant, in our work, the three populations from different sites were investigated. Two of them originated from natural habitats , and one  was collected from Boles\u0142aw waste heap localized at the place of former open-cast mining of Ag-Pb and Zn-Pb ores dating back to the13th century and 18th century, respectively. The analysis of soil from Boles\u0142aw revealed the increased level of Zn, Cd, and Pb  or inhibMicroelements and toxic metals were determined in the investigated plant populations. The results are presented in C. vulgaris showed the ability to accumulate Fe and Cu, especially in the root. The level of Fe reached even to 2000\u2009mg/kg and Cu content was above 50\u2009mg/kg, while the values in the other plants usually ranged from 75 to 400 and from 5 to 20\u2009mg/kg, respectively. In turn, the high variation of heavy metal content between the metallicolous population from Boles\u0142aw (MB) and the nonmetallicolous populations (NN and NP) has been noticed. As expected, the level of Zn, Pb, Cd, Fe, Ni, and Mn was significantly higher in the root and leaves of MB plants as a result of soil contaminations compared to those of the NN and NP. Particularly, high differences were observed for the content of Pb, Zn, and Cd; the concentration was from several to even few hundredfold higher in the MB population. In MB flower heads, the amount of these metals was also increased; however, it was about 10-fold lower than that in MB leaves and roots. The restricted translocation of heavy metals to generative plant organs is common phenomena occurring in numerous species [For most metals, the differences of their content between the populations from natural habitats were only slight or statistically insignificant. The amount of Zn, Cr, Mo, Co, and Mn was in the range typically found in plants . However species . The dif species . The con species . The difMetal ions have a diverse influence on plants. Cadmium and lead belong to the typical toxic components which cause a cellular damage and disturb cellular homeostasis , 19, 20.The excess of metals in a growth environment may cause the adverse process in a plant, such as chlorosis, necrotic leaf spots, and the other morphological alterations. The tolerance on a decreased level of metals is highly dependent on the plant species, cultivars, or genotypes within a species . In our The diameter and number of flower heads and leaves were similar for all tested populations ; however, the other morphological parameters differed significantly. MB plants were about 2-fold lower, and they had shorter and wilder leaves and longer roots compared to reference populations. The differentiation in morphological features between the metallicolous and NN/NP populations is observed by numerous researchers. For example, in plants exposed to Pb chronic stress, roots are usually longer or/and thicker and Pb is cumulated in their outer part to prevent generative organs , 25. TheC. vulgaris were established using a spectrophotometric technique. The comparison of results obtained for the different parts of the plant was presented in Plant and plant-based products may be a rich source of polyphenols, which act as antioxidants, and therefore, they are helpful in preventing oxidative stress . The antC. vulgaris had significant antioxidant activity. The highest ability to scavenge free radical was noted for flower head extracts, and it was more than 2-fold higher compared to that for the leaf extract. In turn, root extracts exhibited the lowest activity, and it may be explained by generally lower production of antioxidants in the underground part of the plant. No or only slight differences between the investigated populations were observed, and it showed that chronic multimetallic stress had no influence on the antioxidant activity. These findings are in accordance with the results obtained by Dresler et al. [r > 0.79, r > 0.61, and r > 0.68, resp.). The detailed correlation data are presented in Table S3.As it can be seen, the aboveground part of r et al. , 25. GenC. vulgaris plant extracts was conducted using an experimentally elaborated gradient elution program which enabled the separation of common plant phenolics [Phenolic acids and their derivatives are considered one of the main groups of plant secondary metabolites with significant antioxidant activity . HPLC ofhenolics . The anay\u2009=\u2009124454737.80\u2009\u00d7\u2009\u221236813, r = 0.9997). The calibration curve was constructed on the basis of the relationship between peak areas and standard concentrations at 5 concentration levels (n = 5).Quantification of chlorogenic acid was performed based on the linear regression equation  of leaf and root data showed a clear separation between the metallicolous and nonmetallicolous populations Figures . It was C. vulgaris is rich in polyphenols and minerals. The species growing in noncontaminated areas contain more chlorogenic acid and possess higher antioxidant activity; thus, these species may become ingredients of herbal teas or natural products where a high amount of phytochemicals and minerals is needed. The results obtained in this study confirm an importance of the plants' growth conditions for the safety and quality of the herbal material.Our results demonstrate that Table S1: Mineralization parameters. Table S2: Validation parameters for AAS analysis. Table S3: Correlation between investigated variables. Figure S1: The diameter and number of flower heads and number of leaves obtained for C. vulgaris populations . Data are means \u00b1 SE. Values followed by the same letters are not significantly different (p < 0.05)."}
+{"text": "This study evaluates andcompares the stepwise pattern of ROS production during in vitro development of reconstructed goat embryos produced by zona-free method of somatic cell nuclear transfer(SCNT). Furthermore, the pattern of ROS production of SCNT embryos were comparedwith zona free embryos derived from in vitro fertilization (IVF).A unique feature of embryo metabolism is production of reactive oxygenspecies (ROS). It is well established that during in vitro development  were used for assessment of ROS production using 2, 7-dichloro dihydroflouresceindiacetate (DCHFDA) probe and the result were presented as fold increase or decreaserelative zona free oocytes.In this experimental study, zona-free oocytes, SCNT and IVFembryos at different stages of The relative level of ROS compared to metaphase-II (MII) oocytes insignificantlydecrease during early stages post embryo reconstitution and regained its value by 8-celland morula stage and, significantly increase compared to MII oocytes by blastocyst stage.The pattern of ROS change in SCNT embryos is similar to zona free IVFderived embryos, except it decrease from two cell stage and regain its value at morulastage. The sudden rise in ROS at blastocyst stage, further emphasizes the special need ofIVF and SCNT derived embryos during this stage of development. SCNT)] can effect ROS generation.Literature study reveal that across all the species studied, including goat, composition of embryo culture media have profound effects on elopment . In regaes (ROS) , 10 and r stages , 11. SCNin vitro effect which it is well established that early embryonic block, and the rise in generation of ROS during IVP are also maternally derived and are independent of paternal contribution , also known as maternal to embryo transition, was first reported by Nasr-Esfahani and Johnson  in mice ribution . During ribution  and Ostaribution  studied This study was approved by the Ethical Committee of Royan Institute. In this experimental study, unless otherwise stated, all chemicals and media used in the present study were obtained from Sigma  and Gibco , respectively.in vitro maturation (IVM) as described previously  containing 10% FCS and antibiotic (1% penicillin-streptomycin) at 37\u00b0C, 5% CO2 in air. Cell started to shed out of the explants. Eventually, these cells proliferate to forms a confluent monolayer within 2-3 weeks. Obtained cells were used for investigation of fibroblast phenotype using differential immunostaining with anti-vimentin (for fibroblasts) and pan-cytokeratin  antibodies  before being cultured in medium that contained 0.5% FCS for 4-5 days. Serum starved cells were subsequently trypsinized and used for SCNT procedure.Ear biopsy of a healthy pre-pubertal female goat was taken, cut into 2-3 mmIn vitro matured oocytes were denuded by vortexing in presence of 300 IU/mL hyalorunidase. Only good quality oocytes with homogenous cytoplasm and extruded first polar body were used for the experiments. The process of zona free enucleation was carried out as described previously by Nasr-Esfahani et al. ]. The reconstructed oocytes were rested for 0.5 hours before being activated using ionomycin  followed by incubation with 2 mM 6-DMAP for 2 hours. Reconstituted-activated oocytes were then cultured in groups of five to seven in modified synthetic oviductal fluid (mSOF) under mineral oil at 38.5\u00b0C, 5% CO2, 5% O2 and humidified air for 7 days in 20 \u03bcl wells.Nuclear transfer (NT) was carried out according to Hosseini et al. . In brie2 in humidified air at 38.5\u00b0C. Twenty-two hours after insemination, cumulus cells attached to oocytes were mechanically removed via pipetting. Then, the zona was removed by brief enzymatic digestion as described above. The presumptive zygotes were then cultured in groups of five to seven as described for SCNT embryos.According to Forouzanfar et al. , maturedThe process of ROS measurement was as described previously . In brieThe relative intensity is defined as the difference in the intensity of embryos from the mean intensity of MII-oocytes/mean intensity of MII-oocytes. It is important to note that for assessment of ROS at each embryonic stage at least three replicates were carried out. For each replicate at least 30 to 80 embryos and 55 to 95 oocytes were assessed.Percentages data were transformed by ArcSin and analyzed by one way ANOVA model of SPSS version 17 . Differences were compared by the Tukey multiple comparison post hoc test. All data are expressed as mean \u00b1 SEM and differences were considered as significant at P<0.05.Figure 1 shows fluorescence images of goat MII-oocyte, zona free SCNT and IVF embryos at different stages of pre-implantation embryo development following staining with DCFHDA for ROS measurement. As depicted, irrespective of embryo production method, fluorescence intensity increased as the embryo progressed toward blastocyst stage.Figure 2 shows the mean relative intensities of embryos relative to MII-oocytes at different stages of development in zona free IVF and SCNT derived embryos. As shown, in zona free IVF embryos, the mean relative ROS levels significantly decreased at 4 and 8- cell stages relative to the mean intensity of MII-oocytes and then began to rise by 16 cell stage which resulted in a significant increases by compact and blastocyst stages relative to all the earlier stages. Moreover, the relative increase at the blastocyst stage was significantly higher than compact embryos.The trend of ROS production in SCNT embryos appear to follow the same trend as those of zona free IVF embryos. In zona free IVF embryos the decrement in ROS occur after 2-cell stage while in zona free SCNT reconstructs, the decrements being at earlier stage (post reconstruction). Subsequently, the increment in ROS production in zona free IVF embryos begin at around 8-cell stage while the increment in zona free SCNT embryo begin at around 4-cell stage. Therefore, despite similar trend of ROS production, a significant difference between the two groups were observed at 2- (lower in SCNT group), 4- (higher in SCNT group) and 8- (higher in SCNT group) cell stages. A significant difference was also observed at blastocyst stage. The degree of ROS production was significantly higher in blastocyst derived from zona free IVF embryos in comparison to zona free SCNT reconstructs.During this study, unlike the zona free IVF embryos, in some of the SCNT embryos one or more blastomeres showed higher fluorescence intensity compared to other blastomeres .Also, it has been reported that some blastomeres, due to asymmetrical division, are non-nucleated. In order to understand if this phenomenon has any relation to intensive ROS levels within blastomeres, the SCNT embryos with non-uniformed ROS staining were also stained with viable chromatin dye  and no relation was observed between ROS intensity with presence or absence of chromatin in each blastomeres. As shown in Figure 3, except for one blastomere without nucleus (arrow), all the other blastomeres with high ROS intensity had nuclei.The results of this study showed that the relative ROS production in zona free IVF derived embryos decreased following fertilization, began to rise at around zygote genomic transition (ZGA) which occur around 8-16 cell stage , and subin vitro embryo development (in vitro development of goat embryos.Although, the pattern of ROS production is species-specific, it has been established that the stage of ZGA in developing embryos of mice and other animals investigated so far, coincides with a sharp increase in ROS level , 20. In elopment . In thisIn accordance with previous reports in other species, , 12, 13,This study for the first time described the pattern of ROS production in reconstructed embryo derived from SCNT procedure and in zona free goat embryo. The results showed two major time points of increased ROS production. The first raise in ROS production was observed during ZGA and the second raise took place during the period of blastocyst formation. These results may emphasize the special need of SCNT and zona free IVF derived embryos to external source of antioxidants during these two critical stages of development which in turn may affect the efficiency of embryo production from these two techniques."}
+{"text": "Oral anticoagulants (OAs) are the recommended drugs to prevent cardiovascular events and recurrence in patients with atrial fibrillation (AF) and cardioembolic stroke. We conducted a literature search to review the current state of OAs pharmacogenomics, focusing on Genome Wide Association Studies (GWAs) in patients treated with vitamin K antagonists (VKAs) and direct oral anticoagulants (DOACs).VKORC1, CYP2C19 and CYP4F2. Variants in these genes have been included in pharmacogenetic algorithms to predict the VKAs dose individually in each patient depending on their genotype and clinical variables.VKAs: Warfarin, acenocoumarol, fluindione and phenprocoumon have long been used, but their interindividual variability and narrow therapeutic/safety ratio makes their dosage difficult. GWAs have been useful in finding genetic variants associated with VKAs response. The main genes involved in VKAs pharmacogenetics are: DOACs: Dabigatran, apixaban, rivaroxaban and edoxaban have been approved for patients with AF. They have stable pharmacokinetics and do not require routine blood checks, thus avoiding most of the drawbacks of VKAs. Except for a GWAs performed in patients treated with dabigatran, there is no Genome Wide pharmacogenomics data for DOACs. Pharmacogenomics could be useful to predict the better clinical response and avoid adverse events in patients treated with anticoagulants, identifying the most appropriate anticoagulant drug for each patient. Current pharmacogenomics data show that the polymorphisms affecting VKAs or DOACs are different, concluding that personalized medicine based on pharmacogenomics could be possible. However, more studies are required to implement personalized medicine in clinical practice with OA and based on pharmacogenetics of DOACs. Oral anticoagulants (OAs) are recommended drugs to reduce the risk of stroke and systemic embolism in patients with non-valvular atrial fibrillation (NVAF), and to treat and reduce the risk of deep venous thrombosis (DVT) and pulmonary embolism (PE) , 2.OAs can be classified as vitamin K antagonists (VKAs)   and direGenome Wide Association Studies (GWAs) have been very successful in finding genetic risk factors associated with complex diseases or with drug response (pharmacogenomics). This technique uses a general approach which allow a systematic agnostic research of common genetic factors across the whole genome , 7.Furthermore, different GWAs analysis have been published describing variants associated with the interindividual and inter-ethnic variation of VKAs response \u201311 and, The aim of this review is to describe the genetic factors associated with the response of OAs drugs used in stroke and other cardiovascular diseases prevention and to discuss the future use of OAs pharmacogenomics in the clinical practice.An extensive literature search was performed, up to December 2017, on PubMed with the following key words: \u2018GWAs and warfarin\u2019, \u2018GWAs and acenocoumarol\u2019, \u2018GWAs and fluindione\u2019, \u2018GWAs and phenprocoumon\u2019, \u2018GWAs and dabigatran\u2019, \u2018GWAs and edoxaban\u2019, \u2018GWAs and rivaroxaban\u2019 and \u2018GWAs and apixaban\u2019. Thirty-eight results were obtained for \u2018GWAs and warfarin\u2019, 6 of which were GWAs studies. For \u2018GWAs and acenocoumarol\u2019, 4 results were obtained. Only 1 of them was a GWAs study performed on acenocoumarol. One result was obtained for \u2018GWAs and phenprocoumon\u2019 which corresponded to a GWAs analysis. Two results were found for \u2018GWAs and dabigatran\u2019; only one was a GWAs analysis in patients treated with dabigatran. For the other searches, no results were obtained Figure .in vitro study for apixaban  the first and second most significant eQTLs genes for the SNPs from the published GWAs cited in this review  inhibitors . They inVKAs have demonstrated to be effective, preventing 15 deaths and 15 non-fatal strokes per 1,000 patients with non-valvular AF. However, they caused 8 non-fatal major extracranial bleeds . VKAs haNormally, during the first days to weeks of treatment, the correct dose is not achieved as measured by the international normalized ratio (INR). INR values above 3 or 4 increase the bleeding risk, whereas INR values below 2 are associated with low efficacy of VKAs . PatientWarfarin is a racemic mixture comprised by S and R enantiomers. The S enantiomer has a half-life of 24-33h, whereas R-warfarin has a half-life of 35-58h . S-warfaThe daily dose of warfarin varies between individuals. Doses range from 2 to 10 mg per day . FurtherRelevant non-genetic factors affecting warfarin dose variance include age, weight, body surface, gender, drug interactions, diseases and the quantity of vitamin K intake .VKORC1) gene, in cytochrome P450 2C9 (CYP2C9) gene and in cytochrome P450 4F2 (CYP4F2) gene ..85].Another meta-analysis including 7 observational real-world studies found similar results . HoweverOther datasets included patients with NVAF treated with dabigatran. All of them observed lower risk for major bleeding, intracranial bleeding and death for patients treated with dabigatran compared to patients treated with warfarin . These aDabigatran 75 mg was approved in the USA for patients with renal impairment. In an observational study, it was observed that the ratio for stroke, mortality and bleeding was similar in patients treated with dabigatran 75 mg and in patients treated with warfarin. However, the ratio for intracranial bleeding was lower for dabigatran patients. However, most of patients included in this study treated with dabigatran 75 mg did not have renal impairment, so the prescription of this drug was off-label , 92.Dabigatran, like the other DOACs, is taken in a fixed dose. However, there are interindividual differences between patients in blood concentration (estimation of 30% variation for systemic exposure) .CES1 was associated with trough concentrations  and with lower risk of any bleeding. The polymorphisms rs4148738 in the ABCB1 gene and rs8192935 in CES1 gene were associated with peak concentrations but not with clinical outcome [ABCB1 and CES1) encode for proteins related to dabigatran pharmacokinetics, which means that variants in genes related to dabigatran absorption and metabolism explain part of the variations in dabigatran concentrations between individuals [The unique GWAs with patients treated with DOACs was developed to identify variants associated with interindividual variability in dabigatran etexilate blood concentration. 1,694 patients of white European ancestry from the RE-LY study treated with dabigatran were analyzed. The polymorphism rs2244613 in the esterase gene  outcome . Howeverividuals .CES1 gene and rs4148738 in ABCB1 gene) to analyze the influence of these polymorphisms on the interindividual variation in dabigatran concentration. They found significant association of the SNP rs8192935 in CES1 gene with dabigatran trough concentration [Another study in 92 patients with AF has recently evaluated the effect of the three polymorphisms found in the dabigatran GWAs analysis  and CYP independent mechanisms , 83, 94.Approximately 1/3 of unchanged rivaroxaban is eliminated by the kidneys. The other 2/3 of rivaroxaban is metabolized by the liver .There are different studies, registries and databases describing the use of rivaroxaban in real-world for patients with AF \u2013103.In a meta-analysis of 9 studies , the res2DS2-VASc  had higher rates of stroke and systemic embolism, major bleeding and death [The Xantus study, a prospective, observational, phase IV analysis, observed that patients with higher CHADS2 and CHAnd death , 100.Different registries and datasets have found results which were similar to the ROCKET-AF phase III clinical trial observations , 99, 101ABCB1 gene or CYP genes could be associated with rivaroxaban concentration in blood.There are no GWAs studies performed in patients treated with rivaroxaban. It is possible that polymorphisms in genes involved in the metabolism of the drug could be associated with rivaroxaban blood levels, similar to the results found with warfarin, acenocoumarol and dabigatran. Taking into consideration the proteins associated with the absorption (P-glycoprotein) and metabolism (CYP enzymes) of rivaroxaban, polymorphisms in Apixaban, like rivaroxaban, is a direct oral inhibitor of the Xa factor. Apixaban is indicated by the EMA and FDA for reducing the risk of stroke and systemic embolism in patients with non-valvular AF. It is used in patients with risk factors  , 2.The bioavailability of apixaban is around 50% and it reaches the maximum concentration 1-3 hours after intake. Its half-life is between 8 and 15 hours. Roughly 25% of the drug is excreted by the kidneys . ApixabaABCB1 gene in apixaban concentrations (the same polymorphism of the dabigatran GWAs) showed an association with apixaban peak concentrations [A genetic study on the effect of rs4148738 polymorphism in trations . HoweverABCB1 gene or CYP genes which metabolize apixaban  could be associated with rivaroxaban blood levels, similar to the results found with warfarin, acenocoumarol or dabigatran.Currently, there are no GWAs studies in patients treated with apixaban. Taking into account the proteins associated with the metabolism of rivaroxaban, it is possible that polymorphisms in the Edoxaban, such as rivaroxaban and apixaban, is a direct oral inhibitor of factor Xa indicated by the FDA for reducing the risk of stroke and systemic embolism in patients with non-valvular AF . EMA hasEdoxaban achieves maximum plasmatic concentrations within 1 to 2 hours after being taken. It has predictable pharmacokinetic profile and 62% oral bioavailability. Edoxaban, as the other DOACs, is also a substrate of the P-glycoprotein. It is metabolized by hydrolysis through a carboxylesterase (CES) and oxidized by the CYP enzyme CYP3A4 . Some 50A meta-analysis of 24 real-world studies included patients with AF to analyzed patients treated with a daily dose of edoxaban 30 mg or 60 mg versus placebo (no treatment), aspirin and aspirin plus clopidogrel. They observed how treatment with edoxaban 30 mg decreased the risk for all stroke, ischemic stroke and mortality. Patients treated with edoxaban 30 mg versus patients treated with aspirin plus clopidogrel had a lower risk of intracranial hemorrhage. Edoxaban 60 mg reduced the risk of any stroke and systemic embolism versus placebo, aspirin and aspirin plus clopidogrel. Thus, both edoxaban doses had a positive net clinical benefit compared to antiplatelet treatment or no treatment in the real world .2 and CHA2DS2-VASc score, both edoxaban doses had higher NCB than warfarin. At CHA2DS2-VASc scores 0 and 1, warfarin had no positive NCB compared to no treatment. In patients with CHADS2 and CHA2DS2-VASc score \u2265 2, edoxaban 60 mg had a better NCB than edoxaban 30 mg and warfarin. At CHA2DS2-VASc scores 0 and 1, edoxaban 30 mg had superior NCB than edoxaban 60 mg. In patients with HAS-BLED score \u2264 2 , both warfarin doses had higher NCB compared to warfarin independently of the CHADS2 and CHA2DS2-VASc scores. In patients with a HAS-BLED score \u2265 3, both edoxaban doses had positive NCB compared to warfarin, in patients with CHADS2 and CHA2DS2-VASc score \u2265 2 [Another study with patients from the Danish nationwide cohort tested the hypothesis that edoxaban had a net clinical benefit (NCB), which is the balance between systemic embolism and intracerebral bleeding, superior to warfarin. Comparing without treatment, warfarin had a NCB of 0.26 prevented events per 100 patients-years, edoxaban 60 mg had a NCB of 0.71 per 100 patients-years and edoxaban 30 mg had a NCB of 0.71 per 100 patients-years. At all CHADScore \u2265 2 .in vitro study was performed with the purpose of determining if two polymorphisms in factor Xa (Ala152Thr and Gly192Arg) were affecting the edoxaban activity. The authors concluded that these mutations do not account for the interindividual variability of edoxaban [One edoxaban .ABCB1 gene, or genes involved in the edoxaban metabolism (CYP3A4 or CES such as dabigatran), and the response to edoxaban could be interesting.There are no GWAs studies performed in patients treated with edoxaban. Studies analyzing the role of CYP2C9, VKORC1 and CYP4F2 are associated with VKAs dose variation between individuals. However, these variants together with non-genetic factors explain about 50% of the interindividual dose variation [Variants in ariation , 30. Furariation . For epiariation , 111.CYP2C9 and VKORC1 together with clinical variables to predict the initial dose of warfarin. The different PAs improved the accuracy and efficiency of warfarin dose initiation. However, the results are controversial in the reduction of out-of-range INRs. Some studies observed that genotyping variants in CYP2C9 improves INR control [VKORC1 have not been genotyped in all studies and could modify the results and conclusions. Probably larger number of patients are necessary to achieve higher statistically power to observe differences among PA and clinical algorithms. Moreover, the genetic variants identified to date explain only a proportion of the drug response, thus more genetic variants can contribute to increasing the percentage of drug variability and may be included in pharmacogenetics algorithms [Diverse pharmacogenetics algorithms have been developed, including polymorphisms in  control , whereas control , 45. Altgorithms .To improve the main shortcomings of VKA, mainly the frequent monitoring, DOACs have been approved for the prevention of stroke recurrence and systemic embolism in patients having suffered a previous stroke with non-valvular AF, to treat DVT or PE. DOACs have a better efficacy-safety profile compared to warfarin (Daiichi Sankyo Europe GmbH 2014). Patients treated with DOACs have a lower ratio of bleeding events  and life-threatening bleeding (dabigatran). Furthermore, the use of DOACs reduces intracranial bleeding compared to warfarin. In patients with AF, the results from the different pivotal clinical trials suggest that DOACs improve the outcomes of these patients . AltogetABCB1, which is involved in the transport of all DOACs, or CYP3A4, involved in the metabolism of all DOACs with the exception of dabigatran [Due to the lack of GWAs with DOACs (only one) , it woulbigatran , 94. FurCYP2C9 and VKORC1 variants can benefit more from edoxaban compared to warfarin [Pharmacogenetics could be useful to identify the more appropriate anticoagulant for each specific patient, avoiding adverse events such as any bleeding or stroke recurrence. VKAs and DOACs do not share the same mechanism of action. This fact distinguishes both kinds of anticoagulant, meaning that VKAs could be more suitable for one individual but DOACs for another. Moreover, VKAs and dabigatran have different metabolic pathways and as different polymorphisms affect metabolism of both kind of drugs, knowing the specific genotype of each individual before starting anticoagulant treatment could help avoid adverse events. For instance, a sub-study of the ENGAGE-AF-TIMI 48 trial demonstrated that patients with the warfarin , 114. ThIt is expected that around year 2020 the results of large randomized clinical trials on the incorporation of pharmacogenetics algorithms to guide VKAs dosage will become available. Furthermore, by this time, it is likely that genotyping will be quickly and more cost-effective. It is expected that the use of pharmacogenetics algorithms for vitamin K inhibitors will reduce adverse reactions to these drugs .Pharmacogenetic studies with DOACs would help to choose the most appropriate anticoagulant treatment for each patient. Thus, pharmacogenetics studies have to be translational, to include genotyping in clinical practice and achieve a more personalized medicine, as happens with other kind of treatments, such as in cancer , 116.TPMT) in acute lymphoblastic leukemia (ALL) patients before treatment with 6-mercaptopurine is useful to select the correct dose based on genetics before the first drug intake. After changing the label for 6-mercaptopurine in 2004, some hospitals routinely started to order TPMT genotyping before initiation of the treatment [The applicability of pharmacogenetics in clinical practice has been mainly demonstrated with cancer treatments. Nowadays, there are cancer drugs which are dosed following pharmacogenetic tests. The genotyping of thiopurine S-methyltransferase (reatment .UGT1A1) have been found to be associated with the risk of developing hematological and/or digestive toxicities. Thus, dose reduction was recommended in patients with the genotype *28/*28. In United States, the FDA recommended genotyped the UGT1A1*28 variant before irinotecan prescription. In Europe, a Dutch workgroup and the French National Thesaurus of Digestive Oncology recommended dose reduction in *28/*28 patients [UGT1A1  [In advanced colorectal cancer, irinotecan is the most widely-used drug. However, polymorphisms in diphosphate-glucuronosyltransferase 1A1 (patients . A genotoratory) , 119.CYP2D6 and tamoxifen for breast cancer, DPYD and fluoropyrimidine antimetabolite 5-fluorouracil (5-FU) for colon cancer, SLCO1B1 and Simvastatin for the reduction of cholesterol levels, HLA-A*33:03 and ticlopidine for secondary prevention of atherothrombosis [Other genes associated with adverse drug events with potential benefits for use in clinical practice could be: rombosis , 120.Current medicine could incorporate pharmacogenetics to offer patients an effective personalized medicine by adjusting treatment and dosage in those cases with a proven genetic association based on GWAs analysis and subsequent approval by the FDA or the European authorities. This way, adverse events related with lack or excess of drug effect could be decreased.This review has some limitations. First, the idea was to revise and analyze all the GWAS published for the different OAs. However, for some of them , there are no studies that have performed GWAS analysis with these drugs. In these cases, other genetic studies based on gene candidates were reviewed. Nevertheless, these kind of studies are biased, as they are based on a previous hypothesis and do not replicate the results in independent populations. Thus, their reliability is limited. It would be necessary to perform a GWAs approach to really identify genes involved in the secondary response to these drugs.in vitro or in silico functional analysis have to be performed to demonstrate the effector gene for each specific mutation.Second, for many polymorphisms found associated with the effect of different OAs is assumed that the effector gene is the closest to the mutations. However, this is not always true and"}
+{"text": "Shittori feel is defined as a texture that is moderately moisturized; however, many people experience \u2018shittori feel\u2019 when they touch a dry solid material containing little liquid. Here, shittori feel was evaluated for 12 materials. We found that the highest score of shittori feel was achieved by powders. Multiple regression analysis showed that shittori feel is a complex sense of moist and smooth feels. We analysed the relationship between the physical properties and the moist/smooth feels to show how subjects felt certain feels simultaneously. The moist and smooth feels are related to the surface roughness and friction characteristics of the materials. The moist and smooth feels can be perceived when the finger starts to move on the material surface and when the finger moves and rubs the material surface, respectively. However, this feel is quite complicated and involves some contradictions. Although this definition includes the word \u2018moisture', many people feel shittori feel when they touch a dry solid material containing little liquid.When we evaluate tactile sense, a contradiction can arise between the definition of a term and the result of sensory evaluation. For example, the Japanese term \u2018turized' . Althougipsticks \u20134. In thipsticks  found thet al. [et al. [In general, the result of sensory evaluation is analysed by tactile dimensions to understand the expression mechanism of tactile feel. Many tactile textures that arise when humans touch objects are described by a combination of simple tactile dimensions. Hollins et al.  evaluate [et al.  conclude [et al. . Takemur [et al.  proposedet al. [et al. [The relationship between the aforementioned tactile dimensions and the physical stimulus applied to the skin, especially friction, is important to not only understand contradictions and complications in tactile sense but also to design cosmetics, clothes, cars and virtual reality systems. Smith et al.  examinedet al. . A corre [et al.  examined [et al.  focused  [et al. . Coated shittori feel is caused by a small variation in the friction coefficient when the subject touches a smooth cold cloth or leather [shittori feel of fabrics were the maximum value of heat flux, mean coefficient of friction, hysteresis of the shear force, shear stiffness and the compression energy.We showed that the tactile feel when we touch the water on the glass surface is due to the periodic fluctuation of the frictional resistance ,16. When leather ,19. The shittori feel even under dry conditions. We first evaluated the phonological impression of the term \u2018shittori feel'. The term \u2018shittori feel' was positioned on a haptic sensory space map. When a word is entered into the system, it is referenced by the phoneme database and auditory impression and is evaluated on the basis of 26 pairs of basic tactile evaluations [shittori feel was evaluated for 12 materials containing powder, artificial leather, resin, metal, silicone rubber or cloth. We also evaluated 10 items with, for example, warm, cold, moist, and dry feels and analysed combinations of tactile dimensions that contribute to the shittori feel. In addition, we characterized the items' physical properties, including their coefficient of friction, surface roughness, surface energy, Young's modulus, thermal conductivity and water content. Because friction affects various dimensions, we imitated the physical conditions when humans touch objects. The surface of the object was rubbed with a contact probe that imitated the surface and mechanical properties and shape of human finger surfaces [shittori feel.We conducted a multifaceted analysis to clarify why humans experience a luations . Next, asurfaces . In addisurfaces ,23. Fina2.2.1.shittori feel' on the Internet. Next, three professional panelists determined 12 objects with various levels of shittori feel: one panelist was a researcher who has been engaged in tactile evaluations for more than 15 years, and the other two were students who have studied tactile sense.2.2.shittori feel was evaluated by a system that can convert a sound-symbolic word in Japanese into quantitative ratios in multiple tactile dimensions (26 pairs of adjectives) [The phonological impression of ectives) . In this2.3Tactile evaluations were performed by 20 female students ranging in age from 20 to 29 years. The evaluations were performed in a quiet room at 25 \u00b1 1\u00b0C and 51 \u00b1 2% in relative humidity after the subjects washed both hands with a commercial liquid hand soap and acclimatized for 20 min. The materials were evaluated in a random order to eliminate order effects, and the subjects touched the materials through a blackout curtain. The purpose of the tests was revealed to the subjects before the evaluations, and the subjects made the decision of whether to join the evaluation by themselves. All evaluations were conducted according to the principles expressed in the Declaration of Helsinki. The responsible party at Yamagata University confirmed that the ethics and safety of the present test were acceptable . Informed consent was obtained from all subjects.shittori feel?' and \u2018Why did you feel that?', respectively. Next, in Q3, the subjects evaluated 10 tactile sensations regarding the tactile dimensions: warm, cold, soft, hard, moist, dry, smooth, sticky, rough and slippery feels [shittori feel was 7 or more, we asked timing when the shittori feel was felt. The choices were as follows: 1, \u2018The moment when you touch the material\u2019; 2, \u2018while touching'; 3, \u2018the moment when you take your finger off\u2019; 4, \u2018I do not know'.When the subject touched solid materials on a table with the forefinger of their dominant hand and moved their finger to the left or right, they evaluated the tactile feel of the material for 25 s . In the ry feels . Finallyry feels . The deg2.41xF and the forces applied to the material 2xF and zF were detected by strain gauge type load cells. The measurement range of the friction forces were as follows: 1xF = 2xF = 0.06\u20139.9 N and zF = 0.06\u20139.8 N. By contrast, the detection limits of 1xF, 2xF, zF are 0.02 N. In this study, 2xF and zF data obtained from the load cells on the sample stage were analysed because an inertial force was applied to the contact probe.The frictional force was evaluated using a sinusoidal motion friction evaluation system in which the contact probe slide on objects under sinusoidal motion was at 25 \u00b1 1\u00b0C and 51 \u00b1 2% relative humidity ,23. ElecV during sinusoidal motion is described by the following equationD, \u03c9 and T are the moving width, angular velocity and time, respectively. The experimental conditions were as follows: \u03c9 = 2.1 rad s\u22121 (maximum velocity = 30 mm s\u22121), vertical load = 0.98 N, D = 30 mm and number of cycle = 11.The contact probe was a finger model made of urethane resin ) . On the The contact probe was in contact with the material for about 30 s prior to sliding. The coefficient of static friction depends on the conditions of friction. In particular, the contact time between the contact probe and the material before sliding can bring a significant effect on the static friction. Therefore, we confirmed that the static friction was almost the same even if the contact time was changed from 10 to 300 s.To confirm the reproducibility, evaluations were conducted three times. The temporal profile of the angular velocity under this experimental condition is shown in electronic supplementary material, figure S2. Data for one round trip from the point where the contact probe changed direction for the first time were selected for quantitative analysis.\u03bcs); (ii) kinetic friction coefficient (\u03bck); (iii) normalized delay time \u03b4 ; (iv) \u03bcs\u2212\u03bck (the difference between static and kinetic friction coefficients); and (v) the friction variation value (the standard deviation of the kinetic friction coefficient)  static friction coefficient . The magnification of the objective lens was 5\u00d7, 10\u00d7 or 20\u00d7. Three different locations on the material surface were observed to evaluate the surface roughness. In the case of powders, the surface was observed after 5 mg of powder was rubbed onto urethane artificial skin  using a sinusoidal motion friction evaluation system. The surface energy was cited from the literature \u201329. The 2.5.shittori feel and tactile dimensions as a target variable and explanatory variables, respectively. Statistical analyses were performed using the Excel version 14.0  and the SPSS 16.0 Base System software . Student's t-test was conducted for each explanatory variable. The null hypothesis was rejected when the p-value was greater than 0.05 [Multiple regression analysis was carried out using han 0.05 .3.3.1shittori. The scores for \u2018moist (wet)', \u2018calm' and \u2018soft' were high: 0.75, 0.66 and 0.56, respectively. The evaluation scores of the tactile dimensions were as follows: moist = 0.75, soft = 0.56, cold = 0.42, smooth = 0.31 and sticky = 0.04. These evaluation results suggest that the shittori feel is a complicated feel that cannot be explained with a single tactile dimension. Although the dominant factor is moist feel, one or more of soft, cold and smooth feels may also affect this sensation.3.2.shittori feel score is shown in A  and B (6.8 \u00b1 1.9), whereas the lowest scores were those for L (1.8 \u00b1 1.5) and K (2.9 \u00b1 2.2). A clear difference was observed between the groups A\u00b7B and K\u00b7L. shittori feel score strongly evoked soft, sticky, moist, smooth and slippery feels. In the comments of Q2 and Q4, the words concerning sticky or moist feel are appeared with relatively high probability for powders A and B. On the other hand, the words related to dry feel were described for cloths K and L with the lowest score of shittori feel .The shittori feel score. As mentioned in electronic supplementary material, table S1, many subjects felt the shittori feel while touching the materials: in the 64 cases with the shittori score of 7 or more, the number that the subjects felt \u2018at the moment when they touch the materials' was 19, while the number \u2018while touching' was 55.By contrast, the scores of soft, warm, rough and dry feels were high when the subjects touched a cloth with the lowest A, B, C and D); group 2 = resins and metals ; and group 3 = silicone rubbers and cloths   . The pow3.3.\u03bcs and \u03bcs\u2212\u03bck between the powders A\u00b7B with a high shittori feel score and the cloths K\u00b7L with a low score. The static friction coefficients of A with the highest shittori feel and L with the lowest were 0.73 and 0.55, respectively; the difference between them was thus 0.18. The parameter \u03bcs\u2212\u03bck was 0.18, 0.15, 0.05 and 0.09 for A, B, K and L, respectively; thus, the value for the powders was more than twice as large as that for the fabrics. These results suggest that the frictional resistance is strong at the start of sliding in the case of a material with a strong shittori feel.Electronic supplementary material, table S4 shows the friction properties, surface roughness, surface energy, Young's modulus, thermal conductivity and the water content of the solid materials. We found a difference in the frictional parameters C\u00b7K\u00b7L, whose surface was covered with fur, and I \u00b7 J, whose surface was uneven: Ra = 44.7, 40.6, 116, 105 and 93.9 \u00b5m for C, I, J, K and L, respectively. The other materials were several to 10 \u00b5m. The surface energy, Young's modulus and thermal conductivity showed large values characteristic of metals. However, no relationship was found between these values and the score of shittori feel. The water content was also high, 7.38 and 6.91%, for fabrics K and L, respectively, although no clear relationship with the fabrics' shittori feel was found.Surface roughness was especially large for 3.4.shittori feel is correlated with many tactile dimensions. The correlation coefficients associated with moist, sticky, smooth, slippery, dry and rough feels were 0.707, 0.477, 0.391, 0.290, \u22120.558 and \u22120.443, respectively. A regression analysis was achieved using shittori feel as a target variable and a combination of moist feel and other tactile dimensions as explanatory variables. Among some regression formulas satisfying conditions of regression analysis, the below equation shows the highest correlation coefficient obtained with the sensory evaluation dataYShittori, XMoist and XSmooth are the score of shittori, moist and smooth feels, respectively.The R for equation (3.1) and the correlation coefficient between explanatory variables (scores of moist and smooth feels) were 0.740 and 0.258, respectively. The significance probability of each explanatory variable p-value was less than 0.05 for moist and smooth feels. shittori, moist and smooth feels. The numerical values in the graph are a score representing the strength of shittori feel. In cases where the score of both moist and smooth feels was higher, the shittori feel became stronger. To perceive the shittori feel, a complex tactile sensation of moist and smooth is necessary.The explanatory variable was moist and smooth feels. The multiple correlation coefficient 3.5.shittori feel. In particular, we wondered why many people perceived a moist feel that is generally thought to be caused by wet conditions even though they touched a dry solid material containing little liquid. Significant correlation indicating that water content and thermal conductivity lead to the enhancement of the moist feel was not observed. However, we found an interesting trend in the friction profile: larger values of \u03bcs\u2212\u03bck are associated with stronger moist feels. \u03bcs\u2212\u03bck. For eight materials except A, H, K and L, a linear relationship between these sensory and physical parameters was observed\u03bcs\u2212\u03bck. The correlation coefficient R for equation (3.2) was 0.918. The significance probability of explanatory variable p-values was less than 0.05 for \u03bcs\u2212\u03bck. However, some exceptions that did not follow this linear relationship were found. For example, metal H and fibres K and L showed moist feels lower than those expected on the basis of \u03bcs\u2212\u03bck. These smaller-than-expected moist feels might be caused by inhibition by a strong dry feel. In response to Q2, 20, 50 and 35% of subjects used words related to the dry feel to describe metal H and fibres K and L. These results suggest that many subjects consciously felt dry feels when they touched these objects. Under such circumstances, subjects are apparently reluctant to give a high evaluation score to moist feel, which is the opposite sense to dry feel. By contrast, powder A showed a moist feel higher than that expected on the basis of \u03bcs\u2212\u03bck. We do not have a clear explanation for this phenomenon. At the present time, no psychological or physical parameters that can rationally explain the high moist feel of powder A have been found.We revealed the physical origin of moist and smooth feels, which are related to the Ra when a 10\u00d7 objective lens was used (correlation coefficient = \u22120.769). Previous studies have suggested that a smooth feel is perceived when the variation of the frictional force with respect to the average frictional force is small [A, B, C, D). In the second group, the surface roughness was less than a few micrometres . These results suggest that there are two types of perceptual processes in the sense of smooth feel.Smooth feel showed a strong negative correlation with surface roughness is small \u201335. Figushittori feel. The shittori feel is perceived through the two senses of moist and smooth feels. In addition, these two senses relate to the surface roughness and friction characteristics of the materials.4.shittori feel even under dry conditions? As mentioned in \u03bcs\u2212\u03bck. The strong resistance applied to the skin surface at the moment of sliding is related to moist feel. This result is consistent with several previous studies. Some researchers have suggested that frictional resistance increases if there is a small amount of water on the skin surface. This increase in friction is caused by the capillary adhesion at liquid bridges between the skin and the object [Fat [A and B, resin E and metal G were dry materials containing almost no water, many subjects can remember the resistant force of a substance containing a little water on the basis of its large friction difference \u03bcs\u2212\u03bck.Why do subjects feel a e object . The macect [Fat . The capet al. [The hypothesis based on the capillary attraction force may be too simple to explain the phenomena that occur on the actual skin surface. For example, the extent of moisture accumulation in the contact region is associated with an increase in the friction . The moret al.  measuredshittori feel. The variation from the moment when the finger touches the object to the end of the finger movement might enable the make perception of complex tactile sensations. The moist feel is perceived the instant the finger starts to move on the material surface, whereas the smooth feel is felt when the finger moves as rubbing the material. First, the moist feel was related to the difference between static and kinetic friction coefficients, suggesting that the moist feel is perceived at the moment when the finger starts to slide because static friction is a force generated when an object begins to move. Next, smooth feel is perceived as the finger rubs the materials, because humans recognize roughness and smoothness through vibrations and deformation of the skin caused by rubbing the surface of the material with their fingers [We consider a mechanism where different tactile feels of moist and smooth are recognized when a subject touches a material with  fingers \u201344.5.shittori feel, which is defined as \u2018a texture that is moderately moisturized\u2019, is a complex sense of moist and smooth feels. The moist and smooth feels relate to the surface roughness and friction characteristics of the material. The moist feel is perceived the instant the finger starts to move on the material surface, whereas the smooth feel is perceived in the process when the finger moves as rubbing the material.We found that shittori feel is an important factor in designing clothes, cosmetics and food in the Japanese market. However, people with different backgrounds did not understand the feeling, so it was a major obstacle in the global company that is developing business in Japan.These findings are important for sharing and using information on tactile sense among people all over the world. It is difficult to understand each other among people with different backgrounds, because the sense strongly depends on language and culture. There is no doubt that the shittori feel by the tactile dimensions and the common parameters such as the friction coefficient will make it possible to design and evaluate products based on this sense all over the world. Moreover, it was confirmed that moist and smooth feels can be explained by friction and surface shape. Interestingly, the relationship between these sensory factors and physical factors was not a simple linear relationship. One of the reasons is that nonlinear mechanical response occurs on soft matter surfaces such as skin. In future, development of a model that accurately describes such a nonlinear response is required.The model that explains the"}
+{"text": "Plasmonic black gold converts CO2 to methane using solar energy. via steam generation. Black gold DPCs also convert CO2 to methane (fuel) at atmospheric pressure and temperature, using solar energy.In this work, we showed the tuning of the catalytic behavior of dendritic plasmonic colloidosomes (DPCs) by plasmonic hotspots. A cycle-by-cycle solution-phase synthetic protocol yielded high-surface-area DPCs by controlled nucleation\u2013growth of gold nanoparticles. These DPCs, which had varying interparticle distances and particle-size distribution, absorb light over the entire visible region as well as in the near-infrared region of the solar spectrum, transforming gold into black gold. They produced intense hotspots of localized electric fields as well as heat, which were quantified and visualized by Raman thermometry and electron energy loss spectroscopy plasmon mapping. These DPCs can be effectively utilized for the oxidation reaction of cinnamyl alcohol using pure oxygen as the oxidant, hydrosilylation of aldehydes, temperature jump assisted protein unfolding and purification of seawater to drinkable water  Plasmonic coupling, a near-field coupling of the dipoles in the presence of light, can also be tuned by modifying the gap/distance between NPs.via \u201cthermal hotspots\u201d and electric fields via \u201cEM hotspots\u201d. The main goal of this work was to establish whether tuning of these hotspots allows tuning of the solar energy harvesting and catalytic activity of these plasmonic colloidosomes.In this work, we developed a novel solution phase synthetic protocol for dendritic plasmonic colloidosomes (DPCs) by controlled nucleation\u2013growth of Au NPs onto high-surface-area dendritic fibrous nanosilica2.2.1via a controlled nucleation\u2013growth synthetic protocol  were used as the deposition site for the Au NPs, and then varying the distances between the Au NPs by using a cycle-by-cycle growth approach protocol .4). To vary the distance between the Au NPs, we decided to increase the size of the Au NP seeds, as with the increase in the size of Au NPs, the distance between them will be reduced (0 species (required for the nucleation and formation of new Au seeds). This was achieved by converting the Au precursor to a less reactive K-gold solution and also by replacing NaBH4 with the weak reducing agent formaldehyde (HCHO). This allowed for the slow formation of Au0 species, avoiding super-saturation, and hence, only the growth of the existing Au seeds in DPC-C0 took place, yielding DPC-C1. By repeating this growth cycle, DPC-C3, DPC-C4, and DPC-C6 were synthesized, with reduced interparticle distances  by reduction with sodium borohydride (NaBH reduced . This waistances . CompletDPCs were characterized by scanning transmission electron microscopy (STEM) for the size and distribution of Au NPs . Energy 3D tomography study of DPC-C3, -C4 and -C6  for 1 h and the temperature of the solution was measured over time  as a temperature-probe reaction.3 and C6 . Howeverd Fig. S6. These r thermal .2.4where Ias is the anti-Stokes signal intensity, Is is the Stokes signal intensity, Vi is the laser frequency (in cm\u20131), Vv is the vibrational mode frequency (in cm\u20131), h is Planck's constant = 6.626176 \u00d7 10\u201334 J s, k is Boltzmann's constant = 1.3807 \u00d7 10 \u201323 J K\u20131, and T is the temperature (in K).To find accurate local-temperature information, SERS-based Raman thermometry was used.x, which was in the range of 60 to 90 \u00b0C, and the local temperature (60 to 155 \u00b0C) at the surface of the Au NPs due to the loss of heat during convection from the Au NP surface to the surrounding environment. Notably, DPC-C3 and DPC-C4 have higher surface temperatures than DPC-C6  hotspots\u201d of the synthesized materials, we performed surface-enhanced Raman spectroscopy,retching . This waretching , due to 2.5The spatial distribution of localized surface plasmon modes was visualized by scanning transmission electron microscopy\u2013electron energy loss spectroscopy (STEM\u2013EELS) plasmon mapping.The increase in crowding as well as the heterogeneity of the Au NPs (reduced interparticle distances) caused more intense hotspot formation, as shown in the 2nd column of 2.62 to methane conversion at atmospheric pressure and temperature using water as the hydrogen source, (iii) protein denaturation studies, and (iv) hydrosilylation of aldehydes.To study the effect of plasmonic hotspots on catalysis and gain mechanistic understanding, we conducted four different studies using these DPCs, (i) oxidation reaction of cinnamyl alcohol using pure oxygen as the oxidant, (ii) CO2 (100 psi), ethanol (5 mL) and reaction time (6 h)  oxidation to cinnamaldehyde (CAL) as a test reaction . For the Table S5. As prevthe dark . A maximthe dark . DPC-C4 the dark . Under dthe dark . Similarnditions  and S8\u2020.x, the COL molecule experiences dramatically amplified EM fields. This facilitates the strong coupling between the plasmons of DPC-Cx and COL molecules, polarizing the COL molecules and weakening the bonds, which in turn enhanced the conversion as well as the kinetics of COL to CAL. Hence, more hotspots resulted in more polarization (EM hotspots) and localized thermal energy.Higher activity under light can be explained based on the varying numbers of EM and thermal hotspots. At the microscopic level, when the COL is positioned within the nano-gap of DPC-Cx can also result in the opening of non-radiative decay pathways like Landau damping which can activate reactants by hot electron injection. 2-dissociation process, via the transfer of an energetic electron from gold to the antibonding O\u2013O 2p*-state of molecular O2 adsorbed on the gold surface. The use of an insulating silica support avoided any back-injection of these hot-electrons, further improving the probability transfer to an oxygen molecule.However, loss of SPR coherence in DPC-CAlso, once hot electrons have undergone the fast electron\u2013electron and electron\u2013phonon scattering processes (10 fs\u201310 ps) to reach thermalization, the absorbed energy is transferred to the localized environment. Since COL oxidation is intrinsically an endothermic reaction, any amount of localized heat will bring about the favorable condition for the reaction to proceed. Thus, the formed highly localized thermal hotspots on the nanoparticles are an excellent source of localized heat that gets transferred to COL molecules in their proximity. These results indicate that the combination of energetic electrons, as well as hotspots , works cooperatively and is responsible for the enhanced photoactivity. Since DPC-C4 possesses optimized hot-spots, it showed the best catalytic activity, over other DPCs.x for CO2 methanation reaction at atmospheric pressure and temperature  protein using DPC-C0, DPC-C1, DPC-C3, and DPC-C6 in the presence of light (400 to 1100 nm). The fluorescence behavior of two tryptophans (Trp-134 and Trp-213) in BSA was investigated by exciting the protein at 280 nm and recording the emission from 300\u2013500 nm. The emission peak maxima for BSA (at 352 nm), on light illumination, due to the thermal effects of plasmonic DPC-Cl change . A maximTo study the explicit role of \u201cEM hotspots\u201d, we chose the hydrosilylation of aldehydes, which does not require energetic electrons . To elim3.In conclusion, we have developed the solution phase synthesis of dendritic plasmonic colloidosomes with varying interparticle distances between gold NPs using a cycle-by-cycle growth approach by optimizing the nucleation\u2013growth step. These DPCs absorbed the entire visible and near-infrared region of solar light, due to interparticle plasmonic coupling as well as the heterogeneity in the Au NP sizes, which transformed gold material to black gold. Raman thermometry and SERS provided information about the thermal and electromagnetic hotspots and local temperatures which were found to be dependent on the interparticle plasmonic coupling. The spatial distribution of the localized surface plasmon modes by STEM\u2013EELS plasmon mapping confirmed the role of the interparticle distances in the SPR of the material.via steam generation. They also catalyzed CO2 to methane (fuel) conversion at atmospheric pressure and temperature, using solar energy.We observed the significant effect of the plasmonic hotspots on the performance of these DPCs for the oxidation reaction of cinnamyl alcohol using pure oxygen as the oxidant, hydrosilylation of aldehydes, temperature jump assisted protein unfolding and purification of seawater to drinkable water x performance. Thus, DPC-Cx catalysts can effectively be utilized as Vis-NIR light photo-catalysts, and the design of new plasmonic nanocatalysts for a wide range of other chemical reactions may be possible using the concept of plasmonic coupling.This was attributed to varying interparticle distances and particle sizes in these dendritic plasmonic colloidosomes. The results indicate the synergistic effects of EM and thermal hotspots as well as hot electrons on DPC-C4.4.1iii) chloride hydrate salt  was added dropwise. Then, the reaction mixture was sonicated for 15 min, followed by stirring for 2 h at room temperature. A freshly prepared NaBH4 solution  was added dropwise to this solution, which was further stirred for 2 h. The resulting material was then isolated by centrifugation, thoroughly washed with water and ethanol, and dried under vacuum at 80 \u00b0C for 12 h. The material was denoted as DPC-C0.Dendritic fibrous nanosilica was functionalized with (3-aminopropyl)triethoxysilane (APTES) by refluxing 4 g of calcined silica with APTES  in 250 mL of toluene at 80 \u00b0C for 24 h. The resulting material was washed repeatedly with toluene and ethanol, followed by drying under vacuum at 60 \u00b0C for 12 h. The obtained material (0.5 g) was dispersed in water (50 mL), and the mixture was sonicated for 15 min followed by 10 min of vigorous stirring at room temperature. To this reaction mixture, gold(4.24 to a 1 mM K2CO3 solution). Then 1 mL of ammonium hydroxide (37%) was added and stirred for 5 minutes. The formaldehyde solution (18 mL) was then added dropwise to the suspension. After stirring for 24 h, the solution was centrifuged at 10\u2009000 rpm and washed three times with deionized water and then ethanol and dried under vacuum at 80 \u00b0C for 12 h. The material was denoted as DPC-C1. These growth cycles were repeated to obtain different samples of DPC-Cx.DPC-C0 seeds (100 mg) were added to a previously prepared 200 mL K-gold solution  theory from N2 physisorption data recorded using a Micromeritics 3Flex analyzer. Approximately 100 mg of each sample was degassed at 120 \u00b0C for 12 h prior to N2 sorption analysis.Scanning transmission electron microscopy (STEM) analysis was carried out on an FEI-TITAN operated at an accelerating voltage of 300 kV. For sample preparation, powders of the DPC-C4.4x into a 10 mL quartz tube. The tube was wrapped with aluminum foil from the top, leaving approximately 3 cm exposed at the bottom, on which light was focused. A thermocouple was then inserted into the tube up to the level of the wrapped foil such that it was not exposed directly to incident light. The sample was then stirred at 400 rpm, and the light was focused onto the tube. The increase in temperature was measured with respect to time for 1 h with a 1 min time interval. The spectral range of the illuminating light was from 400 to 1100 nm with a power of 0.65 W cm\u20132.Elevation in temperature upon light illumination was studied by adding 5 mL of water and 5 mg of DPC-C4.5x (2.5 mg) was taken up in an Eppendorf tube, and 3.5 mL of deionized water was added into it. The Eppendorf tube was vortexed for 30 s, and the solution was transferred to a quartz cuvette. The cuvette was then placed on the weighing balance and tared after stabilization of the placed cuvette. The light was then illuminated from the top of the cuvette, keeping a distance of 2 cm between the cuvette and the light guide. Weight loss, as observed from the balance, was then noted with respect to the time of light exposure. The spectral range of the illuminating light was from 400 to 1100 nm with a power of 0.69 W cm\u20132.DPC-C4.6x material was dispersed into 1 mL of water and sealed with a septum and degassed for 30 min in an argon atmosphere. The ammonia borane complex (100 \u03bcmol) was dissolved in 1 mL of water and injected into the above solution. The sample was then exposed to light with a spectral range from 400 to 1100 nm with a power of 0.55 W cm\u20132 and the evolved hydrogen was monitored using gas chromatography (GC) using a thermal conductivity detector (TCD).In a 50 mL Schlenk tube, 10 mg of the DPC-C4.7\u20131) was the same for each sample. The weight of DPC-Cx needed to make a 1 mg mL\u20131 silica solution was calculated using the Au weight percent obtained from the EDS data. 50 \u03bcL of these sample solutions and 100 mM 4-aminothiophenol ethanol solution were mixed and sonicated for 30 s at room temperature. After sonication, the sample was collected by centrifuging at 8000 rpm for 5 min. The precipitate was washed with 100 \u03bcL of ethanol twice to remove the un-adsorbed chemicals. Finally, the precipitate was collected and dispersed in 25 \u03bcL of ethanol. For performing SERS measurements, a sticker chamber and slide glass were used to dry each sample separately. Raman measurements were performed using an In-via micro Raman setup with a 633 nm laser.All samples were dispersed by sonication in ethanol. To ensure the same particle concentration, the concentration of silica  of both a sample solution and a 0.1 mM 4-ATP solution were mixed and sonicated for 30 s at room temperature. After sonication, each sample solution was dropped onto a cover glass and completely dried at room temperature. For SERS measurements, a sticker chamber and cover glass were used for drying each sample separately. All Raman spectra were acquired using an inverted microscope system  with an objective lens . Spectra were collected with a 0.27 mW 633 nm laser with an acquisition time of 20 s. Each spectrum was collected from different points of the dried and aggregated samples. From one point, the Stokes signal was acquired first. After changing the detection wavelength range (\u223c10 s), the anti-Stokes signal was collected under the same conditions.All samples were dispersed by sonication in ethanol. The concentration of the silica content was fixed at 1 mg mLwhere Ias is the anti-Stokes signal intensity, Is is the Stokes signal intensity, Vi is the laser frequency (in cm\u20131), Vv is the vibrational mode frequency (in cm\u20131), h is Planck's constant = 6.626176 \u00d7 10\u201334 J s, k is Boltzmann's constant = 1.3807 \u00d7 10\u201323 J K\u20131, and T is the temperature (in K).To minimize the effects of heterogeneity in the Au coating, data from eight different points for each sample were collected. Local temperatures were estimated using the following equation  was then added, and the reactor was pressurized with 100 psi oxygen. It was flushed ten times with oxygen. Since the SPR of DPC-Cx ranges from the visible to the NIR region of the spectrum, we used light radiation with a spectral width of 400\u20131100 nm and a light intensity of 1 W cm\u20132. Reaction progress with time was monitored using gas chromatography (GC), and the formed products were identified by mass spectrometry (MS).In a 50 mL quartz tube, 10 mg of the DPC-C2 conversion experiment, 10 mg of the DPC-Cx was taken in sealed glass reactor in 5 mL isopropanol and 0.5 mL water and flushed with CO2 (150 mL min\u20131) for 30 minutes. The reactor was then exposed to light  and methane formation was monitored by online micro-gas chromatography  was placed in a 10 mL round bottom test-tube. Tetrahydrofuran (1 mL), dimethylphenylsilane  and 4-chlorobenzaldehyde  was added and purged with argon gas. The reaction mixture was stirred at RT and the temperature of the reaction was maintained using a water circulator. The reactor was then exposed to light (400 to 1100 nm) and product formation was monitored by gas chromatography-mass spectroscopy.For hydrosilylation of aldehydes, DPC-CThere are no conflicts to declare.Supplementary informationClick here for additional data file.Supplementary movieClick here for additional data file.Supplementary movieClick here for additional data file.Supplementary movieClick here for additional data file.InfographicClick here for additional data file."}
+{"text": "Withania somnifera (L.) Dunal (Solanaceae), known as Indian ginseng or ashwagandha, has been used in Indian Ayurveda for the treatment of a variety of disorders, such as diabetes and reproductive and nervous system disorders. It is particularly used as a general health tonic, analgesic, and sedative. As part of continuing projects to discover unique bioactive natural products from medicinal plants, phytochemical investigation of the roots of W. somnifera combined with a liquid chromatography\u2013mass spectrometry (LC/MS)-based analysis has led to the isolation of two novel phenylpropanoid esters, Withaninsams A (1) and B (2), as an inseparable mixture, along with three known phenolic compounds  and a pyrazole alkaloid (5). The structures of the new compounds were elucidated using a combination of spectroscopic methods, including one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectroscopy (HR-ESIMS). Withaninsams A (1) and B (2) are phenylpropanoid esters that contain a side chain, 4-methyl-1,4-pentanediol unit. To the best of our knowledge, the present study is the first to report on phenylpropanoid esters with 4-methyl-1,4-pentanediol unit. The anti-inflammatory activity of the isolated compounds (1\u20136) was evaluated by determining their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, where compound 3 inhibited LPS-induced NO production (IC50 = 33.3 \u03bcM) and TNF-\u03b1 production, a pro-inflammatory cytokine (IC50 = 40.9 \u03bcM). The anti-inflammatory mechanism through the inhibition of transcriptional iNOS protein expression was confirmed by western blotting experiments for the active compound 3, which showed decreased iNOS protein expression. Withania somnifera (L.) Dunal (Solanaceae), commonly known as Indian ginseng or ashwagandha, is a perennial shrub distributed in India, Morocco, Egypt, Israel, Jordan, South Africa, and the Mediterranean region. Currently, it is cultivated on a small scale in South Korea as well +; HRESIMS (positive mode) m/z:317.1375 [M + Na]+, calcd for C16H22O5Na, 317.1365; UV (MeOH) \u03bbmax nm (log \u03b5): 220 (2.29), 240 (2.10), 290 (2.54), 325 (3.42); IR (KBr) \u03bdmax cm\u22121: 3305, 3126, 1755, 1585, 1492, 1042; 1H  and 13C  NMR spectroscopic data, see White, amorphous powder; ESIMS (positive mode) 4 cells/well) were seeded into a 96-well plate and incubated overnight for adhesion. Following incubation, the cells were treated with compounds for 24 h. Next, Ez-CytoX solution  was added to each well and cells were further incubated for 1 h. The cell viability was assessed by measuring the absorbance at 450 nm.RAW 264.7 cells (6.0 \u00d7 104 cells/well) were seeded into a 96-well plate and incubated overnight for adhesion. Following incubation, the cells were treated with compounds and LPS. After 24 h incubation, supernatants were collected and treated with Griess reagent for evaluating NO concentration in the reactants. The absorbance was measured at 540 nm and NO production was calculated by referring to the nitrite standard curve.RAW 264.7 cells . Each step was followed by washing with 1 \u00d7 PBST 5 times. Briefly, plate was coated with coating antibody solution overnight at 4 \u00b0C and then blocked with 1 \u00d7 assay diluent for 1 h at room temperature (RT). Supernatants were applied to the plate for 2 h at RT and the plate was incubated with biotinylated secondary antibody for 1 h at RT. After reacting the plate with horseradish peroxidase (HRP)-streptavidin for 40 min at RT, the plate was reacted with 4-nitrophenyl phosphate disodium salt in diethanolamine buffer as a substrate for 10 min at dark condition and the reaction was stopped by adding 1 N NaOH. Absorbances of each well at 405 nm were applied to standard curve for calculating the quantity of TNF-\u03b1 at supernatants.5 cells/well) were seeded into 6-well plates and incubated overnight for adhesion. Following incubation, the cells were treated with compounds 3 and 4 in the presence of LPS for 24 h. Total cell lysates were obtained and loaded onto sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteins were subsequently transferred onto nitrocellulose (NC) membrane. Membranes were incubated with primary antibodies against iNOS and GAPDH, following which the membranes were incubated with appropriate secondary antibodies. Finally, the membranes were blotted and protein band intensities were analyzed using Imager 680 .RAW 264.7 cells  and B (2) as an inseparable mixture, along with three known phenolic compounds  and a pyrazole alkaloid (5). Withaninsams A (1) and B (2) are phenylpropanoid esters that contain a side chain, 4-methyl-1,4-pentanediol unit. To the best of our knowledge, the present study is the first to report on phenylpropanoid esters with 4-methyl-1,4-pentanediol unit. All isolated compounds were evaluated for their anti-inflammatory effects on nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages. Compound 3 exhibited NO and TNF-\u03b1 inhibitory properties without cytotoxicity. The active compound 3 inhibited NO production by reducing iNOS protein expression.In the present study, phytochemical analysis of the MeOH extracts of roots of"}
+{"text": "Helicoverpa armigera larvae infected by Helicoverpa armigera nucleopolyhedrovirus (HearNPV) to white (broad-spectrum), blue (450\u2013490 nm), UVA (320\u2013400 nm), and UVB (290\u2013320 nm) lights for the tree-top disease. Our findings suggest that tree-top phenomenon is induced only when the light is applied from above. Blue, white and UVA lights from above induced tree-top disease, causing infected larvae to die in an elevated position compared to those larvae living in the complete dark. In contrast, UVB from above did not induce tree-top disease. Blue light exerted the maximum photo-tactic response, significantly (p < 0.01) higher than white light. The magnitude of the response decreased with decreasing wavelength to UVA, and no response at UVB. Our results suggested that the spectral wavelength of the light has a significant effect on the induction of the tree-top disease in H. armigera third instar larvae infected with HearNPV.Lepidopteran larvae upon infection by baculovirus show positive photo-tactic movement during tree-top disease. In light of many insects exploiting specific spectral information for the different behavioral decision, each spectral wavelength of light is an individual parsimonious candidate for such behavior stimulation. Here, we investigated the responses of third instar  Behavior manipulation in the host-parasite interaction has been defined as the phenotypic changes in the host organism . It has Wipfelkrankheit) where, infected larvae climb high up to the top of the tree before death, resulting in the better dispersal of virus particles with broader foliage cover after liquefaction, thus increasing chances of virus transmission to subsequent host larvae [Baculoviruses are host specific pathogens associated with arthropods and mainly infect lepidopteran larvae . Inducedt larvae ,16.egt) gene of Lymantria dispar multi-nucleopolyhedrovirus (LdMNPV) triggers the tree-top disease in L. dispar larvae. In addition, Autographa californica MNPV was found to induce similar disease in both Trichoplusia ni and Spodoptera exigua larvae but the S. exigua larvae showed a molting dependent effect. Ros, van Houte [S. exigua larvae undergoing molting during infection showed the tree-top disease, and vice versa. They further reported that, the mutant AcMNPV lacking the egt gene did not affect the position of larval death. Similarly, in SeMNPV\u2013S. exigua interaction, egt was involved indirectly in tree-top disease by prolonging the lifespan of infected larvae [egt on the tree-top disease is not conserved among all baculoviruses.Hoover, Grove  reportedan Houte  reportedd larvae . These fS. exigua larvae infected with SeMNPV were positively photo-tactic [Orius sauteri (Poppius) but not Trialeurodes vaporariorum [Tetranychus urticae avoids UVA (320\u2013340 nm) but Aphis fabae is attracted to ultraviolet (UV) light (<400 nm) [Bombyx mori, Callosobruchus maculatus, Dermatophagoides pteronyssinus, D. farina [T. urticae eggs but no any effect from UVA is reported [T. confusum and Drosophila melanogaster [The role of light as a key factor in the induction of tree-top disease is a recently discovered avenue. The o-tactic  with theo-tactic . The varo-tactic ,22,23,24rariorum . Tetrany<400 nm) ,27. The . farina ,29,30. Snogaster . TherefoHelicoverpa armigera H\u00fcbner (Lepidoptera: Noctuidae) infected with HearNPV from pre-symptomatic to tree-top disease stage.For the abovementioned reason, we investigated the effect of different wavelengths of lights categorically: White (broad-spectrum), Blue (450\u2013490 nm), UVA (320\u2013400 nm), and UVB (290\u2013320 nm) in the third instar larvae of Helicoverpa armigera (H\u00fcbner) were reared on a wheat bran rich artificial diet at temperature 28 \u00b1 1 \u00b0C, 50% relative humidity and the photoperiod of 12 L: 12 D (Light: Dark) [H. armigera 4th instar larvae; collected, purified, quantified and stored at 4 \u00b0C as described previously [Larvae of t: Dark) . HearNPVeviously .3) was placed at the bottom of the cylinder. The sides and bottom of the cylinders were wrapped with aluminum foil and placed in the black box in order to block light from other directions. Treatment consisted of 4 different lights: White light (broad-spectrum), Blue light (450\u2013490 nm), UVA (320\u2013400 nm), and UVB (290\u2013320 nm) placed 30 cm above the cylinders for luminescence during light hours of the day (12 L: 12 D). Similarly, treated larvae were kept in a complete dark (0 L: 24 D) as a control treatment. Mock-treated larvae were also subjected to similar treatments. Each experiment was conducted with three independent biological replicates and each replicate consisted of 15 larvae.Newly molted third instar larvae were starved for 24 h and fed with the viral dose of LD90 (2800 OBs/larvae) . InfectiThe vertical position of mock-infected larvae was recorded manually using a measuring scale twice a day till pupation; a red light was used to measure the vertical position of mock-infected larvae in the dark condition. The top vertical position where the infected larva was found dead was recorded at 8-days post-infection (dpi). Death from baculovirus infection was further confirmed by screening for liquefaction of the larvae following death, and those who did not die or survived despite being fed with the virus were excluded from the data analyses.To investigate the effect of direction of light in assay 1, a similar experiment was repeated following the same experimental setup except the aluminum foil on the cylinder was wrapped on the side and the top, excluding the bottom, and all the lights used in the treatments were applied from the bottom at a 30 cm distance from the cylinder placed inside the black box. In addition, control treatment consisted of a similar setup with mock-infected larvae as above. Data were recorded in a similar manner as in assay 1.Data obtained regarding the height of the larvae at death were analyzed using the linear regression model in R (v3.0.0). The regression model was fitted using treatments (different lights) and replications as explanatory variables and the height of death as a dependent variable. Most of the larvae died during its fourth instar so were excluded as an explanatory variable .t-test = 4.163, 7.627, 6.119 respectively; d.f. = 369; p < 0.0001. There was also a significant difference between the height of death of the larvae between the blue light and the white light treatment ; blue light causing the larvae to die at a more elevated position than the white light. Treatment with UVB light however, did not significantly affect the larval position at death compared to that of the complete dark; t-test = 1.728; d.f. = 369; p = 0.0849. No significant differences were found between the replicates   . Mock-in= 0.841) .t-test = \u22123.428, \u22122.852, \u22123.378, \u22123.168 respectively; d.f. = 349; p < 0.001, <0.01, <0.001, <0.001 respectively) . Mock-infected larvae did not show any difference in climbing behavior between treatments, instead, they showed similar patterns of multiple peaks, and all of them descended to the bottom near pupation  . This sipupation .egt), and the need for light from above during a certain time period of infection for the induction of the tree-top disease [H. armigera third instar larvae, and surprisingly found that white light along with blue and UVA applied from above induced tree-top disease, but UVB did not. This striking difference on the wavelength sensitivity and their resultant effect on the photomovement of infected larvae during tree-top disease holds a significant ecological importance because of the fact that viruses are the one that lay behind the strings for this photomovement [The ecological and evolutionary significance of baculovirus induced behavior changes in lepidopteran larva have led to some exciting discoveries adding to the fundamental understanding of the mechanisms involved. Various studies have revealed the involvement of baculovirus gene ( disease ,17,18,20movement ,16 and emovement ,35,36. Amovement  which siOur observations showed that light from above is essential for tree-top disease and this is in accordance with previous findings ,38. Our Drosophila, Euglena gracilis, Cyanobacterium, Frog [Phototaxis mechanism has been observed in many insect species ,41,42,43um, Frog ,49,50,51um, Frog ,52. Therum, Frog .H. armigera larvae. Lights within the spectrum of white, blue and UVA induced the tree-top disease, whereas UVB did not, and the blue light exerted significant photo-movement compared to white light. Therefore, in-depth molecular investigation of the spectral light perception of the infected larvae during the tree-top disease can possibly outline the complete molecular mechanism of the host impetus for the before death climbing behavior.These findings provide a fundamental understanding about the role of different light spectrums on climbing before death behavior of baculovirus-infected lepidopteran larvae. We found that light from above is needed to induce tree-top disease in HearNPV infected"}
+{"text": "Furthermore, an image representing the Myc-YAP colony-formation assay experiment in Fig. 2C was strikingly similar to the data shown for the Control colony-formation assay experiment in Fig. 5B.Oncology Reports has ruled that the article should be retracted from the publication on account of a lack of overall confidence in the presented data. Note that the authors were not in agreement that the errors reported and identified were sufficient to merit the retraction of the article. The Editor apologizes to the readership for any inconvenience caused.The Editorial office subsequently investigated this matter further, and noted that the western blots shown in Fig. 6A and B likewise contained strikingly similar bands that were purportedly showing the results from different experiments. After having considered the various issues that have been brought to light with this paper, together with an appeal from the authors that a Corrigendum be published, the Editor of"}
+{"text": "The unusual superconducting properties of granular aluminum oxide have been recently investigated for application in quantum circuits. However, the intrinsic irregular structure of this material requires a good understanding of the transport mechanisms and, in particular, the effect of disorder, especially when patterned at the nanoscale level. In view of these aspects, electric transport and voltage fluctuations have been investigated on thin-film based granular aluminum oxide nanowires, in the normal state and at temperatures between 8 and 300 K. The nonlinear resistivity and two-level tunneling fluctuators have been observed. Regarding the nature of the noise processes, the experimental findings give a clear indication in favor of a dynamic random resistor network model, rather than the possible existence of a local ordering of magnetic origin. The identification of the charge carrier fluctuations in the normal state of granular aluminum oxide nanowires is very useful for improving the fabrication process and, therefore, reducing the possible sources of decoherence in the superconducting state, where quantum technologies that are based on these nanostructures should work. The nanoscale structure can be advantageous in terms of electric loss, and in the limit of a short constriction can even be used as a non-linear element for quantum circuit design [Granular superconductors have attracted great interest for their rich phase diagram and for practical advantages, such as increased critical field , criticaductance . As a maductance . Such a [x wires ,7. The rt design .Although granular aluminum seems to be a very promising alternative material for qubit architecture, it has a disordered nature, due to a large amount of amorphous aluminum oxide. Previous published structural and morphological studies suggest that the granularity on the size of grains, connectedness, and the intergrain spacing depend on the preparation method , which rx nanowires. The observed phenomenology, although related to the normal state, deserves to be understood in details, since it could also have an impact on the superconducting state, where qubit-based technology operates.In view of all these considerations, DC electric and magneto-transport measurements, as well as voltage-noise investigations, have been performed on AlOc-plane sapphire substrate in a controlled oxygen atmosphere, thus allowing for a wide range of sheet resistances. The mean sheet resistance of the AlOx film was determined to be about 2.2 k\u03a9. However, the sheet resistance can vary up to 50% for different film areas across a 20 mm by 20 mm chip due to inhomogeneities that are related to the sputtering process.All of the investigated wires, as well as the surrounding structures, were made from a single layer of oxidized aluminum. As a first step, a 20 nm thick layer of disordered oxidized (granular) aluminum was grown by reactive sputter deposition on a x structures. The PMMA serves as a sacrificial layer, it, however, also influences (limits) the achievable nanowire resolution. Two anisotropic dry etch (RIE) steps were performed to transfer the structures into the AlOx layer after the resist definition: oxygen RIE for the PMMA, and argon/chlorine RIE for the AlOx.An electron beam lithography process was used to define the wire structures, while using a bilayer of hydrogen silsesquioxane (HSQ) and polymethylmethacrylate (PMMA) resist. Here, the high resolution of the HSQ and the ability to use an organic solvent to lift off the PMMA are beneficial for a clean interface and also to contact the AlOWL) 250, 500, 750, and 1000 nm, respectively. Devices named \u201ci\u201d, instead, are microstrips 5 \u03bcm-long and, nominally, 100 nm-wide.The left panel of A simple two-probe measurement method was employed, since all of the measured resistances were much higher than that of the contact wires to investigate the samples transport and noise properties. Additionally, the wires and contacts contribution to the overall electric noise was verified to be negligible . The valx thin films [RT\u03c1 \u2248 5.2 \u00d7 RT\u03c1  does not change with H for all of the bias currents used , above which they become conducting [E, being computed as the voltage difference divided by the nanowire length. In this framework, the conducting clusters forming the network backbone are separated by insulating regions of very small widths. Through such regions, it is expected that tunneling or hopping conduction will take place, thus providing extra paths for electrons. The appearance of extra parallel conduction channels with the increase in bias, which add to an existing network, does lead to a decrease in the total noise of the network, which is consistent with that observed in Alternative to the magnetic effect, a nonlinear transport regime can also result from a random resistor network (RRN) model and, consequently, percolation processes ,33,34. Tnducting . The macx nanowires can be made by computing the negative reversible CR effect, as:R(E) is the resistance at a fixed electric field and 0R is the resistance value in the linear region. The quantity that is defined with Equation (2) is plotted in E for three nanodevices of different length at the temperature of 9 K. Here, it is possible to identify a nonlinearity threshold at a critical electric field cE \u2248 7.2 \u00d7 \u22121. Similarly, the noise reduction in percentage can be computed as:NL(E) is the noise level value at a fixed electric field and 0NL is the noise level value in the linear region. NR is shown in x nanowires are at cryogenic temperatures. The noise reduction effect is much more evident than the CR effect, revealing a greater sensitivity of the noise spectroscopy with respect to the resistivity measurements. Moreover, the evidence of a saturated noise level at high bias values finds strict analogy with the saturated transport properties that were observed in carbon-wax composites and described in terms of the DRRN theoretical approach [x nanowires.The noise reduction above the onset of nonlinearity has been already observed in conductor-insulator mixtures, such as the carbon-wax system, and, in terms of the DRRN model, has been explained as due to an increase in the total number of fluctuators or, equivalently, in the system size from the fluctuation point of view . A betteapproach . This gix nanowires, not being observed in common nonlinear composites dielectrics. This property of the charge carriers fluctuation mechanisms has a direct impact on the normal state conductance and, as a consequence, on the kinetic inductance of granular aluminum nanodevices, a key requirement for their use in superconducting quantum circuits.Although a nonlinear conducting behavior is expected for metal-dielectric composite nanostructures falling around the percolation threshold ,38, the x nanowires, then it is expected that noise sources can be activated whenever there is an electric field in the material higher than a threshold critical value, and large enough fields can suppress it. In the superconducting state, one could expect that E field is zero inside the nanowire and no noise sources are activated. However, in the presence of RF fields, this is no longer true and the noise sources could play an important role in the dynamics of the circuit attached to these devices. Therefore, the identification and reduction of the charge carrier fluctuations in the normal state, by e.g. improving the fabrication process, could be important for reducing the possible sources of decoherence in the superconducting state.The electric transport and voltage-noise properties of granular aluminum oxide nanowires have been investigated for nanodevices of different width and length and in a temperature range from 8 to 300 K. Differently from thin films, which show a linear ohmic regime, nanowires are characterized by a resistivity reduction with increasing bias. In cycled devices, the negative resistance change is reversible. In this regime, above a critical electric field, the nonlinear current-voltage characteristics are also associated with a strong noise level reduction, whose amplitude exceeds that observed on DC resistance of more than one order of magnitude. This nonlinear electric transport behavior of the nanowires does not seem to have a magnetic origin. Moreover, the magnetic field does not affect the noise amplitude, contrarily to what found in thin films with lower resistivity and at low temperatures. The nonlinear resistance could be explained, instead, in terms of a dynamic random resistor network model, usually considered as a variant of the standard random percolation mechanism. If a DRRN model describes the physics of the AlO"}
+{"text": "Supplemental Digital Content is available in the text Liver fibrosis is a pathological change existing in most chronic liver diseases, which leads to abnormal changes in liver tissue structure and affects the normal physiological function of liver. Without effectively control, liver fibrosis can develop into cirrhosis and increase the risk of liver cancer. Salvianolic acid B  is the main active component in the water-soluble extract from Salvia miltiorrhiza, which is a traditional Chinese medicine usually used for treating cardiovascular and liver diseases. It is reported that Sal B shown a good action against liver fibrosis via numerous signaling pathways, which indicate that Sal B is a potential candidate drug for the treatment of liver fibrosis.We searched the related researches from the following electronic databases: PubMed, EMBASE, Web of science, China National Knowledge Infrastructure (CNKI), China Biology Medicine (CBM), Wan fang Database for Chinese Technical Periodicals and VIP Database. All the databases were searched from inception to December 2019. No restriction of language, publication date, or publication status. PICO of this systematic review are shown as flowing: P, preclinical studies which evaluated the effects of Sal B on the animal models of liver fibrosis with controlled studies; I, received Sal B as only treat in any dose; C, received normal saline, distilled water, or no treatment; O, the primary outcome include measure will be the decrease in liver fibrosis score, and the secondary outcomes include the index of liver fibrosis. All the included data will be analyzed with the software of Review Manager 5.2 and STATA 14.2.The purpose of this study is to conduct a systematic review and meta-analysis to assess the effects on anti-liver fibrosis of Sal B, and this will be contribute to drug development and pathological mechanisms of clinical research.INPLASY202050101, registered on 28/5/2020. In fact, liver fibrosis is a common result of abnormal wound healing of chronic liver injury caused by a variety of etiologies and hoping to be reversed and cured. The annual incidence of chronic hepatitis B progression to cirrhosis is 2% to 10%. Therefore, active treatment of liver fibrosis to reverse or delay its development is of great significance to improve the quality of life of patients and improve the prognosis of the disease.Liver fibrosis is a pathological change existing in most chronic liver diseases, mainly manifested as the excessive hyperplasia and deposition of the extracellular matrix (ECM) in liver tissues, which leads to abnormal changes in liver tissue structure and affects the normal physiological function of liver. If treated timely, liver fibrosis and early stage cirrhosis can be reversed.,7 However, the effect of western medicine treatment of liver fibrosis is not ideal because it lead to a series of adverse reactions, coupled with the complex pathophysiological of liver fibrosis, there is still lacks effective western medicine in clinical.,9 It is noteworthy that the active ingredients in Traditional Chinese Medicine are reported to have anti-hepatic fibrosis effects.At present, the treatment of liver fibrosis mainly includes eliminating stimulation or harmful factors, inhibiting liver inflammation, interfering with the activation of stellate cells, and promoting the deterioration of ECM.,12 Sal B have various pharmacological effects, such as antagonizing atherosclerosis, alleviating myocardial ischemia/reperfusion injury, anti-inflammation and alleviation liver injury. It is reported that Sal B shown a good action against liver fibrosis in animal model experiment via numerous signaling pathways,,15 which indicate that Sal B is a potential candidate drug for the treatment of liver fibrosis. However, the efficacy of Sal B for liver fibrosis have not been systematically evaluated.Salvianolic acid B  is the main active component in the water-soluble extract from Salvia miltiorrhiza, which is a traditional Chinese medicine usually used for treating cardiovascular and liver diseases.To be the best knowledge, the purpose of this study is to conduct a systematic review and meta-analysis to assess the effects on anti-liver fibrosis of Sal B, and this will be contribute to drug development and pathological mechanisms of clinical research.22.1 and the recommendations for reporting of systematic reviews and meta-analyses of animal experiments.,18 The protocol was registered on International Platform of Registered Systematic Review and Meta-analysis Protocols (INPLASY202050101).The review authors conducted this systematic review and meta-analysis protocol according to the Preferred Reporting Items for Systematic Review and Meta-analysis Protocol Guidelines (PRISMA-P)2.22.2.1The systematic review will include preclinical studies which evaluated the effects of Sal B on the animal models of liver fibrosis with controlled studies. No restriction of language, date, or publication status.2.2.2We will include the animal models of liver fibrosis and which histopathology proved that there were liver fibrosis in animal model.2.2.3The intervention group will include animals that treatment with Sal B  and received Sal B as only treat in any dose, the Sal B have been administered during or after the induced of liver fibrosis animal models.2.2.4The comparison group will include animals that interventions with normal saline, distilled water, or no treatment.2.2.5Primary outcome include measure will be the decrease in Liver Fibrosis Score, evaluated through histopathologic examination, and measured at the last time following administration of the intervention. Secondary outcomes include the index of liver fibrosis, such as HA, LN, PCIII, Hyp; index of liver function, such as ALT, AST, ALB, TBIL; indicators of oxidative stress, such as SOD, MDA, and GSH.2.3In this systematic review, vitro experiment, abstract of congress, letter to the editor, and human studies will be excluded; Sal B conjunction with other compounds or Sal B based prescriptions will be excluded because effect size may not be due to salvianolic acid B. Non-liver Fibrosis model, no control group, duplicate publications, and no available data also will be excluded.2.42.4.1The systematic review will searched from the following electronic databases: PubMed, EMBASE, Web of science, China National Knowledge Infrastructure (CNKI), China Biology Medicine (CBM), Wan fang Database for Chinese Technical Periodicals, and VIP Database. All the databases were searched from inception to December 2019. No restriction of language, publication date, or publication status.2.4.2We will screen the sites of animal research organizations, Google Scholar, Baidu Scholar. In manual search, 2 reviewers will examine the reference list or citations found in the secondary studies to validate and identify possible eligible studies.The search strategy will use a combination of MESH terms and free terms. Details of the PubMed search strategy appear in Supplementary Materials 1.2.5All the studies will imported to the EndNote software of version X9. Two independent authors will screen the titles and abstracts. The duplicates studies will be removed during the screening process. Disputes between authors will be discussed and resolved by group discussion or third review. After initial screen and de-duplication, the same 2 independent authors will estimate the full text of studies according to the included and excluded criteria. For the duplicate publication studies, we will select the studies which with more complete data. The screening process was shown as Figure 2.6The same 2 independent authors extracted the data following details from included studies:1.the first authors name and publication year, model of liver fibrosis, and the anesthesia methods for model preparation;2.the specific information of animals for each study, including animal species, number, sex, and weight;3.the treatment groups information, including therapeutic drug dosage, method of administration, duration of treatment, and the same information of control group;4.the outcomes\u2019 data of mean value and standard deviation were extracted from every study, including liver fibrosis score, index of liver fibrosis  and index of liver function , index of oxidative stress , and timing for outcome assessments.The data of highest dose was included when the treatment group included various doses of the target drug. The result of the peak time point was included when the data were expressed at different times.2.7The methodological quality of the included studies will be assessed according to the SY stematic Review Centre for Laboratory animal Experimentation SYRCLEs risk of bias tool, and the Collaborative Approach to Meta-Analysis and Review of Animal Data from Experimental Studies (CAMARADES) checklist that evaluate the included studies, 1 point was given for each of the following criteria:1.peer reviewed publication;2.control of temperature;3.random allocation to treatment or control;4.blinded induction of model;5.blinded assessment of outcome;6.use of anesthetic without significant intrinsic neuroprotective activity;7.animal model ;8.sample size calculation;9.compliance with animal welfare regulations;10.statement of potential conflict of interests.2.8All the included data will be analyzed with Review Manager 5.2 version software . The continuous or dichotomous type of date will be included in this review, and the estimate of the continuous date combined effect sizes will calculate by the standardized mean difference (SMD), Dichotomous data will be calculated by the odds ratio (OR), 95% confidence intervals (CIs) of all results were calculated.2.9I2 statistics, when I2 statistics is higher than 50%, indicates a high heterogeneity, random effects model will be used for meta-analysis; otherwise fixed effects model will be carried out. If there is high heterogeneity, we will find the potential sources of heterogeneity by using the methods of sensitivity analysis and subgroup analysis. If the data is not suitable for pooled or subgroup analysis, we will conducted a narrative description analysis of the findings. If the data are available, subgroup analysis will be performed with the following groups: animal species, the method of animal model induced, and the duration of treatment.The heterogeneity will be checked by 2.10The publication bias will be evaluated  by using the funnel plots combine with statistical test (Egger and Begg test), and the Egger and Begg test will be performed by STATA software with version 14.2 .3The purpose of this systematic review and meta-analysis is to provide preclinical research evidence and existing relevant evidence regarding Sal B to improve the fibrosis degree in animal models of liver fibrosis. At the same time, this study aim to show the advantages and limitations of the current literature, and put forward the future research prospects in this field.Although many experimental studies on the effect of Sal B on liver fibrosis have been published, but there is no consensus in the literature. Therefore, it is necessary to conduct a systematic review of the existing experimental studies.Xiaocong Ma designed this research and drafted the manuscript, Meiwen Tang tested the feasibility of the study, Liying Lu and Jinghui Zheng contributed to the development of the selection of studies and data extraction, Jingjing Huang contributed to risk of bias and quality assessment, Junhong Li conducted the data analysis and Weisheng Luo read, provided feedback and approved the final manuscript. All authors approved the final version of the manuscript."}
+{"text": "Patients receiving a solid organ transplantation, such as a kidney, liver, or lung transplantation, inevitably have to take drugs to suppress the immune system in order to prevent rejection of the transplanted organ. However, these drugs are known to cause malignancies in the long term. This study focuses specifically on newly developed carcinomas in patients who use those drugs after a solid organ transplantation. This systematic review and meta-analysis of published data show a 20-fold risk to develop a carcinoma after solid organ transplantation compared to the general population, with specifically increased risks in patients who receive cyclosporine or azathioprine. By comparing the different pathways involved in immunosuppression and the occurrence of carcinoma development, new insights can be discovered for future research and understanding of carcinoma development in transplantation patients and the general population as well.Immunosuppressive therapy after solid organ transplantation leads to the development of cancer in many recipients. Analysis of the occurrence of different types of de novo carcinomas in relation to specific immunosuppressive drugs may give insight into their carcinogenic process and carcinogenesis in general. Therefore, a systematic search was performed in Embase and PubMed. Studies describing over five de novo carcinomas in patients using immunosuppressive drugs after solid organ transplantation were included. Incidence per 1000 person-years was calculated with DerSimonian\u2013Laird random effects model and odds ratio for developing carcinomas with the Mantel\u2013Haenszel test. Following review of 5606 papers by title and abstract, a meta-analysis was conducted of 82 studies. The incidence rate of de novo carcinomas was 8.41. Patients receiving cyclosporine developed more de novo carcinomas compared to tacrolimus  and mycophenolate . Patients receiving azathioprine had higher odds to develop de novo carcinomas compared to mycophenolate  and head and neck carcinoma compared to tacrolimus . To conclude, patients receiving immunosuppressive drugs after solid organ transplantation have almost a 20-fold increased likelihood of developing carcinomas, with the highest likelihood for patients receiving cyclosporine A and azathioprine. Looking into altered immune pathways affected by immunosuppressive drugs might lead to better understanding of carcinogenesis in general. Solid organ transplantation patients receive different immunosuppressive drugs to prevent graft rejection. Each of these drugs inhibits the immune system in a specific manner. Calcineurin inhibitors, such as cyclosporin A (CsA) and tacrolimus (TAC), inhibit the proliferation of T cells which is important to prevent graft rejection ,2. AnothEven though outcomes of solid organ transplantation have improved dramatically since the discovery of immunosuppressive drugs, their use comes with a drawback. Overall, a two to seven times higher risk for development of de novo malignancies can be found in transplant recipients compared to the general population ,8. Long-This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) . A literData of the included articles were extracted using a standardized data extraction form, including study design, patient demographics, duration of follow-up, number of transplant recipients, transplantation period, and number of patients with de novo carcinomas. Corresponding authors were contacted by email regarding missing follow-up data by E.Z. De novo head and neck carcinomas were defined as ear, nose, pharynx, larynx, lip, oral (gland), buccal, tongue, or tonsil carcinomas. Likewise, de novo colorectal carcinoma was defined as colon and rectal carcinomas. De novo uterine carcinoma included uterus and cervix carcinomas. The types of immunosuppressive drugs recorded for the included articles were AZA, CsA, MMF, TAC, SIR, and EVER.To assess the quality of the included articles, the Newcastle\u2013Ottawa Scale was consulted as risk of bias tool . The codp value less than or equal to 0.05 was considered statistically significant. All possible comparisons of immunosuppressive drugs present in the included cohorts were tested. Outcomes of comparisons with two or more study cohorts were considered eligible.The incidence of de novo carcinomas per 1000 person-years was calculated and pooled with DerSimonian\u2013Laird random effects model in RevMan 5 . The oddAfter duplicate removal, the search identified 6318 records. Based on title and abstract, 5569 records were excluded. Consequently, 749 full-text articles were assessed for eligibility. After exclusion of 667 articles, a total of 82 were included for qualitative and quantitative synthesis .n = 11), followed by France (n = 9), Italy (n = 7), and Korea (n = 6) (n = 64) were hospital-based, while others were database-guided or multicenter studies. Thirty-two authors were contacted regarding missing follow-up data, of which only 3 replied and 10 had invalid contact information.Overall, these 82 studies comprised a total of 237,540 recipients, who received 207,304 kidney, 21,404 liver, 5865 heart, and 2235 lung transplants. Transplant recipients were followed up for a mean period of 84.8 months after transplantation. Patients were diagnosed with a de novo carcinoma at a mean age of 52.3 years and after 66.8 months of follow-up. The baseline characteristics of the included cohorts are presented in  (n = 6) . The vasOverall, 33 of the 82 studies scored 5 or 6 out of a maximum of 8 points, which represents a fair quality. A score of 5 or 6 was mainly due to missing information regarding follow-up in the outcome category and the ascertainment of exposure in the selection category. The remaining 49 articles were considered high-quality studies with a score equal to or over 7 .p < 0.00001). De novo carcinoma occurrence in the included studies varied from 7.81 to 115.4 cases per 1000 person-years. Patients who underwent a heart transplantation developed more de novo carcinomas compared to kidney and liver transplantations, particularly de novo bladder and upper gastrointestinal tract carcinomas  (p = 0.59), yet there appeared to be a trend towards higher occurrence of de novo esophageal and duodenal carcinoma in patients who received CsA  (p = 0.03) (p = 0.10). There was no significant difference between patients using CsA and SIR, an mTOR inhibitor   . No diffctively) . Signifi = 0.03) . There w = 0.87) .p = 0.03). Furthermore, there appeared to be a trend for a higher overall likelihood of developing a de novo carcinoma and for developing de novo lung carcinoma   (p = 0.01)  . Patient = 0.01) .p = 0.33)  .This systematic review shows that patients who receive immunosuppressive drugs after solid organ transplantation have a high incidence of de novo carcinomas, with an almost 20-fold increase compared to the age-corrected general population, as indicated by the WHO Global incidence of cancer between ages 30 and 69 (0.43 cases per 1000 person-years) . This agThe current meta-analysis did not show a significant correlation between specific carcinomas and different immunosuppressive drugs, except for significantly less head and neck carcinomas in patients using the calcineurin inhibitor TAC or the antimetabolite MMF compared to the antimetabolite AZA. However, the comparison between MMF and AZA has to be interpreted carefully, as it was only described in two of the included studies, which consisted of unequal cohorts of patients. Many of the comparisons were rarely described in the included articles, which might cause those comparisons to be underpowered. Therefore, the lack of statistically significant results for those evaluations should not be considered irrelevant, but should warrant future epidemiological studies.Furthermore, in agreement with previously published studies, the current results show that overall de novo carcinomas occur more often in solid organ transplant recipients using the calcineurin inhibitor CsA compared to MMF and TAC ,19. For Calcineurin inhibitors CsA and TAC are considered to have a similar working mechanism on the immune system via the calcineurin pathway. The main working mechanism is inhibition of the calcineurin activity in immune cells, thereby preventing the activation and nuclear translocation of nuclear factor of activated T-cells (NFAT), leading to inhibition of Interleukin-2 (IL-2) production in T cells . IL-2 isBoth CsA and TAC promote tumor formation by inducing tumor growth factor-\u03b2 (TGF-\u03b2) and inhibiting apoptosis and DNA repair. This results in enhanced growth and diminished apoptosis of cancer cells ,26,27,28Additionally, in vivo CsA has been shown to induce tumor progression and angiogenesis, independent of calcineurin, by releasing mitochondrial reactive oxygen species, leading to stimulation of mitogenic pathways in tumor and stromal cells . AnotherMoreover, patients using CsA also have a higher incidence of Kaposi sarcoma and lymphoma ,34. TakiBoth CsA and AZA give a higher odds to develop de novo carcinomas compared to MMF, even though MMF is more potent in preventing graft rejection than AZA ,35. BothThe final group of drugs in this meta-analysis are the mTOR inhibitors. Aside from lymphocytes proliferation inhibition, mTOR inhibitors play a role in the intracellular signaling pathways in all cells of the immune system. For instance, the lifespan of and expression of costimulatory molecules by dendritic cells are increased, while on the other hand, metabolic NK cell function is reduced by mTOR inhibition .As many malignancies upregulate the mTOR pathway, the mTOR inhibitors are currently used as anticancer therapeutics . In thisIn the first months after transplantation, the risk of graft rejection is highest. Therefore, patients receive induction therapy in the first period after solid organ transplantation. In the included articles, the induction therapy regimen is scarcely described. Each center decides the optimal regimen based on the patient\u2019s characteristics, but induction therapy mostly consists of a triple therapy combination of corticosteroids, IL-2 receptor antagonists, polyclonal antilymphocyte and antithymocyte preparations, and monoclonal antibody targeting. The effect of solely the induction therapy on carcinogenesis is still unknown. A Cochrane review from 2017 regarding the polyclonal and monoclonal antibody therapies showed an uncertain effect on malignancies . It has Within the immunosuppressive regimen, patients often switch to different drugs in case of chronic rejection, adverse events, or the availability of new drugs. Four years after the transplantation, less than half of the patients still used the first prescribed combination . HoweverThere are many other factors contributing to the carcinogenesis process, such as smoking, alcohol, diet, and genetics. For certain carcinomas, including HCC and cervix carcinomas, viruses can also play a pivotal role in the carcinogenesis process. For HCC, hepatitis B virus (HBV) causes a 100-fold increase of the relative risk to develop HCC. The oncogenic role of HBV is not completely understood as it might be caused by both direct and indirect mechanisms, including immune-mediated hepatic inflammation leading to genetic damage, the induction of oxidative stress, and integration of the HBV DNA into the host genome that induces chromosomal instability . Due to Most studies had a long inclusion period, which might lead to differences in treatment regimens. The longest study had an inclusion period of 47 years and in another study, the first included patient received their solid organ transplantation in 1963. Since then, a lot has changed in the knowledge and possibilities of immunosuppressive drugs. A major breakthrough was the discovery of cyclosporine A, which was first given to patients in 1978 . FurtherThis systematic review has several limitations. Many of the cohort studies included were conducted retrospectively, based on small groups of transplant recipients, and were often hospital-based. Small cohorts might lead to overestimation of the effect. Even though articles with less than five de novo carcinomas were excluded, this might still have introduced some overestimation. Using the NOS score, most articles were deemed to be of fair quality. One of the major problems was lacking information regarding follow-up data. Even though all authors were contacted regarding missing follow-up data, this might still introduce some risk of bias as not all authors replied to supply the follow-up data. These missing follow-up data also limit direct comparability of calculated incidences to the general population. However, the clear trend towards a higher incidence of carcinoma after solid organ transplantation cannot be ignored. Furthermore, large (inter-)national registries and studies based on International Classification of Diseases (ICD) codes might have missed a few de novo carcinomas as diagnosis might not always be coded correctly in the patient records. This could lead to underrepresentation of de novo carcinoma occurrence and bias in the outcome category. Additionally, changes in the immunosuppressive drug regimens could have been missed. A thorough check through each individual patient record is the only way to prevent missing changes in therapy leading to ascertainment bias and missing de novo carcinomas leading to assessment of exposure bias. Many studies were not eligible due to the strict requirements that the immunosuppressive drug regimen and the total solid organ transplantation group described. Finally, the changes in maintenance therapies were rarely described, while this might also influence the cancer development in the long term.To determine the specific correlation between immunosuppressive drugs and cancer development, a combination of a large prospective cohort with sufficient follow-up for carcinomas to develop and translational research is needed. Important confounders can be determined from the prospective cohorts and further examined in in vitro and in vivo models.Looking in depth into pathways of calcineurin inhibitors, such as IL-2, TGF-B, and HIF1\u03b1, and antimetabolites pathways may lead to enhanced comprehension of carcinogenesis in transplant recipients. Additionally, as described, these pathways are also contributing to carcinogenesis in the general population. Exploring these pathways would thus be an interesting topic for translational research and could in the long term give rise to preventive and therapeutic options for specific types of cancer, both in patients who underwent a solid organ transplantation and in the general population.This systematic review and meta-analysis show an almost 20-fold higher likelihood of de novo carcinoma development in patients using immunosuppressive drugs after solid organ transplantation. The likelihood is highest for patients receiving cyclosporine A and azathioprine. By looking in depth into the pathways affected by these immunosuppressive drugs, a deeper understanding of carcinogenesis can be achieved and new starting points for translational and clinical research might be found."}
+{"text": "It has been argued that infants possess a rich, sophisticated theory of mind (ToM) that is only revealed with tasks based on spontaneous responses. A mature (ToM) implies the understanding that mental states are person specific. Previous studies on infants\u2019 understanding of motivational mental states, such as goals and preferences have revealed that, by 9 months of age, infants do not generalize these motivational mental states across agents. However, it remains to be determined if infants also perceive epistemic states as person specific. Therefore, the goal of the present study was to use a switch agent paradigm with the classic false belief violation-of-expectation task. Results revealed that 16-month-old infants attributed true and false beliefs to a na\u00efve agent \u2013 they did not perceive beliefs as person specific. These findings indicate that the mechanisms that underlie infants\u2019 implicit attribution of beliefs differ from those assumed for explicit reasoning about beliefs. The depth of infants\u2019 theory of mind (ToM) is currently the subject of a heated debate. For decades, researchers have attempted to determine exactly when this foundational socio-cognitive ability develops. Traditionally, ToM was thought to emerge between 3 and 5 years of age . Over thToM is defined as the understanding that oneself and others have mental states that guide our behaviors . As suchThere are several proposals that posit leaner interpretations of behaviors observed with implicit ToM paradigms see . If thisanimate objects, but to agents, which are objects that display cues such as autonomous motion, goal-oriented, contingent, and action-at-a-distance behaviors . Therefore, if infants reason about beliefs with the mechanisms attributed to older children, then we expect no group differences in looking time. However, if infants\u2019 looking patterns replicate those found in a priori power analysis required 48 infant participants per belief condition to obtain a moderate-strong effect size (d = 0.90) and adequate power (1 \u2212 \u03b2 = 0.85). This target effect size was taken from previous research . Infants were randomly assigned to one of two conditions: congruent (n = 25) or incongruent (n = 25). Eight additional infants were tested and excluded from the analyses due to fussiness during the task administration.The sample was composed of 50 infants . Infants were randomly assigned to one of two conditions: congruent (n = 27) or incongruent (n = 27). Five additional infants were tested and excluded from the analyses due to fussiness during the task administration.The sample comprised 54 infants  have been published . Infants were randomly assigned to the congruent (n = 23) or incongruent (n = 25) groups.A control condition was used to compare this switch-agent false belief condition with a same-agent false belief condition . This sample was previously tested in our laboratory and data from a subsample  were recruited. Thirty adults  were also recruited on a university campus in a large Canadian city. Participants were students enrolled in Psychology (13), Natural Sciences (11), Business/Finance (4), and Exercise Science (2).To verify how older children and adults process the task, 40 preschoolers . In these cases, the caregivers wore a sleep mask over their eyes to avoid biasing the infants\u2019 looking behavior.The task was administered on a stage-like apparatus (107 \u00d7 97 \u00d7 104 cm). This apparatus had a back wall (107 \u00d7 97 cm) that was separated in four small doors (the right top and bottom doors: 56.5 \u00d7 43.5 cm and the left top and bottom doors: 55 \u00d7 43.5 cm). As in As in the original version of this task , infantsThe first familiarization trial (12 s) began with the screen rising to reveal E1 sitting (at eye level with the infants) behind two boxes (a yellow box and a green box) and a small cup placed on the table in between the two boxes see . E1 raistrue belief that the cup was located in the yellow box). Once this infant-directed pause ended and the curtain was lowered, E1 and E2 switched position, such that E1 was now raising/lowering the curtain and E2 was the agent in the scene. When the test trial (6 s) began, the curtain was raised to reveal E2 sitting behind the two boxes. E2 raised her head for a brief moment to ensure the infants noted the change of agent. Infants in the congruent group saw E2 reach in the yellow box (congruent with E1\u2019s belief) and infants in the incongruent group saw E2 reach in the green box (incongruent with E1\u2019s belief). E2 paused with her hand inside the box until the end of the infant-directed pause. The third experimenter live-coded the infant\u2019s looking time at the scene to transition to the next trial after the infant-directed pauses. Infants\u2019 total looking time (in seconds) at the scene during the infant-directed test pauses was recorded. The waving pre-trial and agent-switch in the test trial excluded, this paradigm was an exact replication of In the belief-induction trial (8 s), the two bottom doors behind the two boxes were closed such that E1 was now standing behind these doors watching the toy cup move from the green box to the yellow box. The toy cup changed location without the involvement of E1, who observed the change of location in this scene while E2 moved the toy cup using the magnet from under the stage. Once the toy cup was inside the yellow box, the infant-directed pause began, during which E1 kept her gaze on the yellow box . The infant-directed pause started once the cup enters the green box. Again, once the screen was lowered at the end of the belief-induction trial, E1 and E2 switched positions. When the test trial (6 s) began, the curtain was raised to reveal E2 positioned behind the two boxes. As in the true belief condition, E2 raised her head for a brief moment then reached either in the yellow box (congruent condition) or in the green box (incongruent condition). Again, infants\u2019 total looking time (in seconds) at the scene during the infant-directed test pauses was recorded by a third experimenter.Infants in the false belief condition saw the same waving pre-trial and three familiarization trials as in the true belief condition. During the belief-induction trial (24 s), infants also saw E1 watch as the toy cup moved to the yellow box see . HoweverThe children completed the task at the end of a testing session including other tasks, and received a small gift and a certificate of merit. The parent received $20 to cover travel expenses. Before viewing the pre-recorded video, adult participants completed a demographic questionnaire and were entered in a draw with the possibility of winning a $20 prize. The video consisted of the exposure trial, three familiarization trials, and the induction trial of the false belief switch-agent condition. The video was interrupted after the curtain was raised at the onset of the test trial, showing the na\u00efve agent sitting behind the two boxes. Adults and children were asked the following question \u201cDo you think the actor will search in the yellow box or in the green box?\u201dFor the VOE task, Habit 2000 was used to live-code the infants\u2019 looking time during the infant-directed pauses. To obtain a more precise measurement, infants\u2019 looking time was recoded offline using INTERACT 8.0 (Mangold) by the primary experimenter. To assess reliability, a second coder who was blind to the hypothesis of the study coded 25% of the video recordings. Cohen\u2019s kappa reliability was 0.85 for the false belief videos and 0.82 for the true belief videos.Using z-scores with cut-offs of \u00b13.0, one participant\u2019s response in the test trial of the congruent condition in the true belief condition and one in the test trial of the congruent condition in the false belief condition were identified as an outlier. These scores were replaced with the next highest value within 3 SDs of the congruent condition mean. Following this modification, the distribution of infants\u2019 looking time at the screen during all trials was normally distributed. Analyses conducted with the samples that included these outlier scores yielded the same results.SD = 9.09) whereas infants in the congruent group looked for 17.46 s (SD = 9.80). A 2 (side) \u00d7 4 (pairing) ANOVA was conducted to determine if the infants developed an agent/color preference during the waving pre-trial \u2013 the side variable refers to infants\u2019 looking time to each side of the stage during this trial given that the position of the two agents was counterbalanced across infants. No main effect of side  or pairing  nor an interaction  were found, indicating that infants looked equally to both agents across all four pairings. In other words, no agent, color, or side preference was found. Next, a 3  \u00d7 2 (group) ANOVA was used to analyze whether infants in the two groups differed in their pattern of looking during the familiarization trials. A significant main effect of trial was found . No main effect of group  nor an interaction  were observed. Planned comparisons indicated that infants looked longer during the first familiarization trial  than during the second  and third familiarization trials . Further, infants looked longer during the second familiarization trial than during the third familiarization trial . On average, infants in the incongruent group looked at the scene for 10.97 s (SD = 5.35 s) during the familiarization trials and those in the congruent group looked for 9.06 s (SD = 3.37 s).First, analyses were conducted to make sure that the infants looked at both agents during the waving pre-trial. On average, infants in the incongruent group looked at the scene during this trial for 20.18 s (SD = 10.50) while infants in the congruent group looked for 17.87 s (SD = 10.13). Once more, a 2 (side) \u00d7 4 (pairing) ANOVA was conducted to determine if the infants developed an agent/color preference during the waving pre-trial. No main effect of side  or pairing  nor an interaction  were found. This indicated that there was no agent, color, or side preference during the waving pre-trial. A 3  \u00d7 2 (group) ANOVA was conducted to determine if infants\u2019 looking during the familiarization trials differed across the two groups. As in the true belief condition, a significant main effect of trial was found . No main effect of group  nor an interaction  were found. Planned comparisons indicated that infants looked longer during the first familiarization trial  than during the second  and third familiarization trials . There was no difference between infants\u2019 looking in the second and third familiarization trials . On average, infants in the incongruent group looked at the scene for 11.51 s (SD = 5.14 s) during the familiarization trials and those in the congruent group looked for 11.06 s (SD = 3.41 s).On average, infants in the incongruent group looked at the scene during the waving pre-trial for 19.33 s  = 11.58, p = 0.001, \u03b72 = 0.10] and a main effect of group . Post hoc analyses revealed that infants looked longer in the false belief condition  than in the true belief condition . Further, across conditions, infants in the incongruent group looked longer  than those in the congruent group . Given that the hypothesis predicted longer looking time for the incongruent than congruent condition, planned comparisons were conducted. In the true belief condition, infants in the incongruent group  looked longer than those in the congruent group  during the test trial . In addition, in the false belief condition, infants in the incongruent group  looked longer than the congruent group  during the test trial , where infants in the switch-agent condition  looked longer than the infants in the same-agent condition . The ANOVA also revealed a main effect of group , where infants in the incongruent group  looked longer than the infants in the congruent group  across both conditions.A 2 (Agent Condition: Same\u2010 vs. Switch-Agent) \u00d7 2 (Group: Congruent vs. Incongruent) ANOVA revealed a main effect of agent condition [n = 16) than for the green box (n = 14). These predictions are not different than what would be expected from chance . Similarly, preschoolers also predicted that the agent was equally likely to reach for the yellow box (n = 19) or the green box .Adults predicted that the agent was equally likely to reach for the yellow box , they should conclude that the novel agent possesses no belief  about the object\u2019s location. This is what adults and preschoolers expected when we asked them to predict where the na\u00efve agent would search for the object in the false belief condition. They correctly assumed that without some previous access to objects or events, beliefs cannot be formed unless through interactions with a knowledgeable agent \u2013 beliefs are person specific. Of course, the explicit prediction required by the task that they completed was not equivalent to the VOE task and future research should aim to examine how preschoolers and adults behave in the traditional VOE design.full-fledged understanding of belief remains to be determined. One could speculate that it coincides with the emergence of explicit belief reasoning during the preschool years and future research will be required to address this issue . Such interpretation is compatible with recent findings showing that 16-month-olds generalized false belief to a toy crane . Althouggoals to the ignorant agent, challenging that infants understand that goals  are person specific. Further, there are mixed reviews regarding infants understanding of the binding properties of preferences  over another (congruent); they were not, however, surprised that the ignorant agent was searching at all. Thus, it appears that infants also generalized the knowledgeable agent\u2019s es i.e., , indicates i.e., . In anotes i.e., . Infantses i.e., . Future Both true and false belief conditions were administered in the present study. The true belief condition permitted an assessment of the seeing = knowing hypothesis recently brought forward by It is possible that infants believed that they were in a situation of natural pedagogy so that all information they were shown is generalizable to all observers. By the display of ostensive cues (looking at and waving at the infant) during the exposure phase, it could be argued that infants developed the \u201cexpectation that the content of the demonstration represents shared cultural knowledge and is generalizable along some relevant dimension to other objects, other occasions or other individuals\u201d , p. 1150reason about other individual\u2019s mental states  broadly to all agents as well as across agents. Nevertheless, the present findings could also be in line with In sum, the present study provides additional evidence that the rich, mentalistic view of ToM understanding in infancy should be toned down. Instead, it appears that infants are, in fact, attributing mental states to agents, but too broadly for this ability to be considered as \u201csophisticated\u201d as in older children and adults. Such broad attribution of mental states is likely adaptive for younger infants, but as they develop, they gradually form more sophisticated understanding of mental states, starting with goals and preferences, and eventually beliefs as children enter the preschool years.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Concordia University Human Research Ethics Committee. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.All three authors conceptualized the study. CN and KB tested the participants, coded, and analyzed the data. KB wrote a first draft of the manuscript that was edited by CN and DP-D. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."}
+{"text": "To identify indicators of guideline-concordant care in lung cancer, to implement such indicators with cancer registry data linked to health databases, and to pilot them in a cohort of patients from the cancer registry of the Milan Province.Thirty-four indicators were selected by revision of main guidelines by cancer epidemiologists, and then evaluated by a multidisciplinary panel of clinicians involved in lung cancer care and working on the pathway of lung cancer diagnosis and treatment in the Lombardy region, Italy. With a modified Delphi method, they assessed for each indicator the content validity as a quality measure of the care pathway, the degree of modifiability from the health professional, and the relevance to the health professional. Feasibility was assessed using the cancer registry and the routine health records of the Lombardy region. Feasible indicators were then calculated in the cohort of lung cancer patients diagnosed in 2007\u20132012 derived from the cancer registry of the Milan Province. Criterion validity was assessed reviewing clinical records of a random sample of 114 patients (threshold for acceptable discordance \u226420%). Finally, reliability was evaluated at the provider level.Initially, 34 indicators were proposed for evaluation in the first Delphi round. Of the finally 22 selected indicators, 3 were not feasible because the required information was actually not available. The remaining 19 were calculated on the pilot cohort. After assessment of criterion validity (3 eliminated), 16 indicators were retained in the final set and evaluated for reliability.The developed and piloted set of indicators is now available to implement and monitor, over time, quality initiatives for lung cancer care in the studied health system.The online version contains supplementary material available at 10.1186/s13690-021-00528-0. Lung cancer remains a killer disease being the leading cause of cancer death for both genders in countries with a high Human Development Index (HDI), with world age-standardized mortality rates of 36.4 per 100,000 for males and 14.6 per 100,000 for females, accounting for approximately 22% of all cancer deaths in 2018 [There is accumulating evidence that patients not receiving guideline concordant treatments have worse outcomes , 4. HencThe aim of this project was to develop, with a multidisciplinary panel, a set of adherence to guidelines indicators concerning aspect of diagnosis, treatment and follow-up of lung cancer care and to pilot them in a cohort of lung cancer patients from the Cancer Registry of the Milan Province, a territory covered by the Agency for Health Protection of Milan, Italy.eSurv (https://esurv.org/). The survey consisted of five sections evaluating the following dimensions: organization, diagnosis, surgery, medical treatment and follow-up. Each indicator was presented with a title and a description of how the indicator would have been calculated from the register and the administrative data. Below it was asked to rate, using a seven-point Likert scale, the validity of the indicator as a quality of care measure, the possibility to modify the value of the indicator at the patient level by the health professional, and the usefulness of the indicator for self-assessment. Definitions provided to the panel are provided in English translation in the We first identified relevant evidence-based practice guidelines and potential indicators from the literature and then, with a modified Delphi method, used clinical experts\u2019 knowledge to select the final set of indicators in two rounds. The whole process is summarized in Supplementary Figure We calculated all the finally selected indicators on a pilot cohort derived from Cancer Registry of the Agency for Health Protection of Milan, Italy including all patients developing a lung cancer  topographic codes C33\u201334) in the period 2007\u20132012 and registered with the Lombardy Regional Health Service. A subset of the area covered by the Agency was used, corresponding to fourteen municipalities around Milan with a total population of 1.546.237 inhabitants at 01/01/2013 . The canDescriptive statistics including quartiles were calculated for the validity, modifiability and utility scores received by each indicator in the first Delphi round. Differences in the distribution of covariates across gender, stage or years were assessed using the Manthel-Haenszel test, analysis of variance or Wilcoxon test, as appropriate. All tests were two-sided with a significance level of 0.05. We calculated each indicator as the proportion of patients who received the procedure in the defined time window among those defined as eligible. Indicators were calculated also stratified by age , sex, Charlson index  and stage (I to IV). To assess reliability, we measured noise as the variance of a proportion, and signal using the variance of the random effect of a hierarchical logistic model with the indicator value as the outcome, age, gender, stage and Charlson index as first level predictors, and provider as the only second level variable. All analyses were performed with SAS software .n\u2009=\u2009532 were excluded as not relevant and n\u2009=\u20098 because they were guidelines already included in the SAGE database search results. Three manuscripts were excluded on the basis of the full-text evaluation [n\u2009=\u200934 indicators, each assessing a recommendation, was included in the web-based questionnaire for the first round of the Delphi process.The search of the SAGE clinical guidelines database retrieved 45 records ; 85 questionnaires were completed in full. Thirteen indicators did not satisfy the selection criteria and were excluded from the second round. One additional indicator evaluating time from thorax computed tomography (CT) to surgery was proposed by panel members (O4). Four indicators were partially modified based on the comments received in round 1. The meeting constituting the second round led to approval of the changes in the indicator formulation but did not modify the number of indicators. Table\u00a0n\u2009=\u200988) and mesenchymal histology (n\u2009=\u200926) cases were excluded. This left n\u2009=\u20095746 patients, 91% NSCLC and 9% SCLC (Supplementary figure p\u2009=\u20090.009), while there were no significant changes in age distribution (p\u2009=\u20090.8). Overall 43% of patients had a Charlson comorbidity index greater than zero, with males having more comorbidities . No differences in Charlson index distribution were observed with stage (p\u2009=\u20090.5).In the 2007\u20132012 period there were 5860 cases of lung cancer  topographic codes C33\u201334) in the resident population of the Milan province. Death certificate only  and were eliminated Table\u00a0: \u2018TreatmCalculation of the organizational indicators showed that the proportion of patients having rapid access to treatment after first contact or diagnostic procedures did not differ between men and women and was higher in younger, healthier and more advanced stage patients O1-O4, Table\u00a0, Table\u00a04This article describes the development, based on the evidence based guidelines and expert consensus, of a set of indicators aimed at monitoring concordance of lung cancer therapy to guidelines and its piloting on 5746 lung cancer patients diagnosed over a 6-year period.One of the major strengths of this work is the participation of a high number of clinicians involved in the different steps of lung cancer care, guaranteeing that the identified reference guidelines were endorsed and perceived as relevant. Secondly, identifying the cohort with a cancer register allows to adequately define the denominators of the indicators, solving the problem of accurately identify the target population that has been encountered using administrative records only . Also, lLung tumors still have a poor overall prognosis, which is certainly related to the often advanced stage at diagnosis . HoweverWith the exception of S1 and S6, which are short term outcome-indicators, it was decided to focus on process indicators measuring adherence to guidelines. Process indicators have been criticized as they do not directly measure the outcome , 47, whiThe use of the current administrative databases offers the possibility to implement the indicator on a population scale and to repeat the measurements in time at a reduced cost. However, it introduces some limitations. The first one is that we were not able to calculate three of the indicators produced by the Delphi consensus process because the investigated procedures were not coded in the necessary databases. We also had to drop three indicators having more than 20% discordances between reviewed clinical records and indicators calculated from the administrative databases. However, this is an important information to plan quality improvement initiatives for the future, raising the problem to consistently employ the codes with the clinicians involved in lung cancer care. Also, when using administrative data to calculate health indicators and define case-mix variables (e.g. comorbidity index), concerns about data quality are always present , 50, 51,The developed and piloted set of indicators is now available to implement and monitor, over time, quality initiatives for lung cancer care in the studied area.Additional file 1: Supplementary methods.Supplementary Figure 1. Process from literature review to indicator calculation and evaluation. Supplementary Figure 2. Selection of the pilot cohort of lung cancer patients from the Cancer Register of the Province of XX."}
+{"text": "Magnetic two-dimensional (2D) van der Waals materials have attracted tremendous attention because of their high potential in spintronics. In particular, the quantum anomalous Hall (QAH) effect in magnetic 2D layers shows a very promising prospect for hosting Majorana zero modes at the topologically protected edge states in proximity to superconductors. However, the QAH effect has not yet been experimentally realized in monolayer systems to date. In this work, we study the electronic structures and topological properties of the 2D ferromagnetic transition-metal dichalcogenides (TMD) monolayer  Topological phase has been one of the main themes in solid-state physics and materials science in the past decade. Especially, topological materials have\u2014in view of their robust topological surface or edge states\u2014shown great promise in spintronics. Since the study of graphene, Haldane shows that the Landau level can be presented in graphene if including an external term  such as sulators  and Bi2T [Bi2Te3 ), electr [Bi2Te3 ), ion doe3 [WTe2 ), and ex [Bi2Se3 , HgSe mo [Bi2Se3 ), has opIn particular, the quantum anomalous Hall (QAH) effect, which presents topological properties in time-reversal symmetry breaking systems, has been reported in y)2\u2212xTe3 , MnBi2TeMnBi2Te4 ,11, and TbMn6Sn6 . MoreoveTbMn6Sn6 . HoweverNovel magnetic two-dimensional (2D) materials, another widely studied class of layered materials bounded by van der Waals force, have attracted increasing attention in recent years owing to the great potential for spintronic applications. A number of monolayer magnetic materials, such as VSe2 ,15, CrI3e2 [CrI3 , CrGeTe3 [CrGeTe3, and Fe3Fe3GeTe2  have beerie=470K  above roow 130 K ,15,19. TIn this work, we study the topological properties, Hall conductivity, and spin Hall conductivity in magnetic TMD monolayer The first-principles calculations of monolayer e (VASP) ,21 basede (VASP)  generalie (VASP)  hybrid enterface . The bano method .To carry out the Wilson loop ,27 and H\u03d5  by(1)\u03d5mmth band at mth band, Here, E and temperature T is just the The intrinsic Hall conductivity one (BZ) , that isHere = 3.36 \u00c5 ,30, the The exchange-correlation potential and band gap are usually underestimated in DFT simulations , includiK points in graphene [Xene monolayer [graphene  and Xeneonolayer .To study the topological phase of monolayer To show the topological edge states on the (010) boundary of the To detect the edge states of two-dimensional materials is a great challenge in experiments. However, the Hall conductivity and spin Hall conductivity measurements serve as a much more viable approach for evidencing the edge states. The intrinsic Hall conductivity can be obtained from the Hall conductivity of spin up and down channels viaHere the In this work, we focus on the ferromagnetic phase of monolayer rie=470K  while abCDW=130K . TherefoThe calculated intrinsic Hall conductivity of Equation ) over thins zero c. Based This relation presents a clear route to experimentally examine the topological properties of the 2D TMD material A large number of recent studies have demonstrated various classes of topological phases with magnetism both experimentally ,12,41,42 1T\u2212VSe2 ,51, but [1H\u2212VSe2  shows thIn summary, we propose that evel see b. This ioperties ,54,55. Poperties . This isrie\u22437.5K . Our stu"}
+{"text": "Corporal punishment (CP) is still a common practice in schools globally. Although illegal, studies in South Africa report its continued use, but only a few have explored factors associated with school CP. Moreover, extant studies have not shown the interrelationships between explanatory factors. This study aimed to determine the prevalence and factors associated with learners\u2019 experiences, and to examine pathways to the learners\u2019 experiences of CP at school.3743 grade 8 learners (2118 girls and 1625 boys) from 24 selected public schools in Tshwane, South Africa, enrolled in a cluster randomised controlled trial evaluating a multi-component school-based intervention to prevent intimate partner violence, and completed self-administered questionnaires. We carried out descriptive analysis, simple linear and structural equation modelling to examine factors and pathways to the learners\u2019 experience of CP at school.About 52% of learners had experienced CP at school in the last 6 months. It was higher among boys compared to girls. Experience of CP at school amongst learners was associated with learner behavior, home environment, and school environment. Learners from households with low-socio economic status (SES) had an increased risk of CP experience at school. Amongst boys, low family SES status was associated with a negative home environment and had a direct negative impact on a learner\u2019s mental health, directly associated with misbehavior.CP in public schools in South Africa continues despite legislation prohibiting its use. While addressing learner behaviour is critical, evidence-based interventions addressing home and school environment are needed to change the culture among teachers of using corporal punishment to discipline adolescents and inculcate one that promotes positive discipline. While the use of a physical methods of discipline, also referred to as corporal punishment (CP) at school is banned and no longer an issue in European countries; it continues to be prevalent in a third of the world\u2019s countries, despite evidence regarding its harmful physical, mental and behavioural effects on the child , 2. The South Africa is a highly inequitable society and this inequality is reflected in the historically complex two-tier education system namely, the middle class, private schools and the public schools . The midStudies have been conducted to understand the reasons underlying the continued use of CP in schools in South Africa. CP is administered for both serious and minor offences such as being absent from school or not doing homework, not knowing the answer to the questions asked by teachers in class, caring a gun at school, and for talking or disrupting a lesson in class , 11. ManWhile previous research has focused on determining the prevalence of learner experiences of CP in schools, limited research has focused on understanding what increases the risk of individual learners experiencing CP at school in South Africa. To contribute to this knowledge gap, we present analysis of quantitative baseline data collected from grade 8 learners who enrolled in a Skhokho Supporting Success randomised controlled trial (RCT) , 15. TheEvidence from South Africa and elsewhere suggests that school CP is undergirded by a myriad of individual, school, family and broader community level risk factors , 16. In Family background, in particular, low socio-economic status is associated with experience of CP at school . ChildreThere are broader community and societal influences to children\u2019s experiences of CP. In communities where use of harsh discipline strategies is acceptable and where CP is perceived as effective in controlling behaviour, learners tend to be at increased risk of experiencing CP , 24. LeaThe Skhokho Supporting Success primary prevention study was a cluster randomized controlled trial (RCT) that sought to develop and evaluate a multi-faceted school-based interventions to prevent intimate partner violence , 15. TheSchools were selected if they were enrolling Grade 8 learners in 2014. All Grade 8 learners in 24 purposively selected English medium State secondary schools within a 50km radius from Pretoria City were eligible to participate in the study. While most (21 of 24) schools were located in black townships including Mabopane, Soshanguve, Hammanskraal, and Mamelodi, Winterveld and Atteridgeville, two were located in the central business district of Pretoria and one in Laudium, a predominantly Indian community . ParticiThe South African Medical Research Council Ethics Committee gave ethics approval for the trial. Permission to work in schools was given by the Gauteng Department of Basic Education (DBE), at the provincial and district level. Permission was also granted by the school principals in the 24 schools. The information letter detailing the nature and purpose of the study, the interventions, participants, what the project would offer to schools, risks and benefits of participating in the study, project timelines, and rights of participants was provided to principals. School principals were also informed that all data generated from the study will be kept confidential and that research reports and articles which will be submitted in scientific journals will not include any information that may identify the school or any of the educators, learners, school governing body officials and parents. All Grade 8s from the 24 schools were eligible to participate in the trial. Participants (learners) gave written assent and consent was obtained from their parents or caregivers before they were enrolled in the trial. All learners in Grade 8 in the participating schools were given a pocket booklet with phone numbers of local professional services which offer help on psychosocial, violence and substance abuse and other challenges. Any learner who specifically identified him or herself as in need of help during the study period was, with her consent, taken by teachers to a relevant local service, as per DBE standard practice. There was no financial re-imbursement for participating in the trial. The trial is registered on ClinicalTrials.gov as NCT02349321. The combined total school enrolment was 6076 of which 4095 (67.4%) obtained consent from the parents or caregivers to participate. Of the 4095, 3811 (93.1%) provided assent and participated in the trial. The percentage of learners in each school that did not complete the survey (incomplete surveys) ranged from 0.6% to 7.0% .A survey was conducted using a self-administered questionnaire loaded in personal digital assistants (PDAs). The questionnaires were presented in English and two local languages spoken in the study area, SeTswana and SePedi. A participant could choose to use any of the three languages, either through text or voice to complete the questionnaire. Participants privately completed the survey in class and could ask for help from trained fieldworkers who oversaw the data collection process. After the survey, the data were uploaded to a secure server only accessible to the investigators and downloaded for cleaning and analysis , 15.This paper presents an analysis from the baseline data from the Skhokho. The binary outcome is the learner\u2019s experience of CP at school, derived from responses to one item: \u2018In the past 6 months were you ever beaten by a teacher?\u2019, with a \u2018yes or no\u2019 response.Our hypothesized conceptual model looks at the inter-relationship between a learner\u2019s home environment, mental health, attitude to schoolwork, learner behavior, and experience of CP at school. These factors are derived from several measures as summarized in We used baseline data from the trial, and we applied data analytical procedures which took into account the school clustered sampling design and the cross-sectional nature of the baseline survey. This was an analysis of the baseline data from a cluster RCT design, with participants clustered within schools. We carried out descriptive analysis on all potential explanatory factors associated with learner\u2019s experience of CP at school as described in our conceptual model. We used logistic regression to assess the relationship between experience of CP at school and all individual measures described in The analyses were done separately for boys and girls and the final structural models are presented in Figs .The total number of learners enrolled in grade 8 in the 24 schools in 2014 when recruitment was done was 6076. A total of 4095 learners had parental consent to participate in the study, and 3811 gave learner assent, agreeing to participate. Of the 3811, 3743 completed the baseline survey: 2118 girls (56.6%) and 1625 boys (43.4%). Fourteen percent of boys and six percent of girls were 15 years and older. Most of the learners (69%) lived in a brick house and almost half (46%) lived with both biological parents. One in seven learners (14.5%) did not live with their biological parent. Forty-seven percent of the learners were living with an unemployed caregiver. More boys (47%) than girls (39%) belonged to a club or society. Most girls (90%) were actively involved in church compared to 85% of boys. Sixty percent of learners had ever dated, and six percent had used alcohol and drugs. Boys were more likely to have dated, to use alcohol or other substances One in five learners (20.6%) had repeated a grade, and boys (28.1%) were more likely to have done so compared to girls (14.9%). Fifty-two percent of learners had experienced CP from teachers at school, and 44.8% from parents or caregivers at home in the last 6 months with levels of experience higher among boys compared to girls. More than half (60.5%) of boys had experienced CP at school and 48.3% at home, while 46.3% of girls experienced CP at school and 42.3% at home in the 6 months preceding the interview.Table 4 shows the pathways leading to boys\u2019 experience of CP at school. The structural equation model shows a direct effect of family SES on the learner\u2019s home environment, with low family SES status associated with negative home environment . Low family SES status also has direct negative impact on a learner\u2019s mental health . The home environment has direct impact on learner\u2019s mental health  and their behavior at school (to school work or towards educators or peers) . The model also shows a significant impact of learner\u2019s mental health on their general risky behavior  (\u03b2 = 0.15). For the boys, their misbehavior affects their relationship with their parents or caregivers at home . As hypothesized, learner\u2019s experience of CP at school is influenced mainly by their school environment  .Table 4 show the pathways leading to girls\u2019 experience of CP at school. The pathways are similar to boys as was hypothesized but with some minor differences. For girls, despite showing negative impact of low family SES on mental health, this relationship was not significant. But consistent with boys, there was a strong relationship between home environment and learner\u2019s mental health , learner\u2019s school climate  , and their misbehavior . Boys mental health has impact on their general misbehavior  . As hypothesized and consistent with boys, girls\u2019 experience of CP at school is influenced mainly by their school environment  .This study aimed to describe the prevalence, factors associated with, and pathways to experiencing CP at school in the past six months. We found a high prevalence of CP experienced by learners at school, with more than half (52%) of the learners reporting CP experience in the past six months. The findings of this study highlight a slightly higher prevalence compared to that found in a national study (49.8%) conducted in 2012 , and conOur data has shown that the risk factors associated with learner experience of CP at school includes learner\u2019s behaviour, home environment, school climate, and other factors including family\u2019s SES and learner\u2019s mental health. For learners, irrespective of gender, school environment  has a direct association with learner experience of CP. Learners\u2019 negative perceptions towards teacher behaviour were associated with experiencing CP at school. Current evidence suggests that this relationship could go either direction. For example, some studies have shown that, for learners, having negative perceptions towards teacher behaviour increases the risk of experiencing CP at school, whereas others have shown that CP at school leads to learners\u2019 negative perception towards teacher behaviour , 37. TheEven though in the bivariate analysis we found an association between sexual behaviour, substance use, bullying and CP at school; overall, no direct association was found between learner behaviour and CP at school in the multivariate analysis. For boys, learner behaviour had direct effects on home and school environment while among the girls, learner behaviour had direct effects on school environment only. School environment was the key factor associated with experience of CP for boys and girls, and this relationship was mediated by other factors including learner behavior and home environment. Addressing learner experience of CP at school requires that we not only focus on learner behaviour, but also on the home and school environments which have a direct association. As hypothesized, our data have shown that the home environment  has an indirect relationship with experience of CP at school, mediated by the school environment. Amongst girls, the home environment has a direct association with learner behaviour, which has an influence on the school environment, and experience of CP at school. This finding can be explained by the gender role expectations amongst girls compared to boys in some South African homes . Some caStudies have shown that boys are more likely to misbehave which results in intolerance and negative caregiver and teacher attitude towards them , 43. WitFurthermore, our findings have shown that the home environment was directly associated with learner\u2019s mental health and learner behavior amongst girls. Learners who experience lack of care and support, and neglect at home are likely to experience poor mental health, which contributes to learner misbehaviour. Our study has shown that the influence of home environment and mental health also connects through family SES amongst boys. A gender effect of socio-economic status was also observed in other studies . InterveThe study had limitations in that the findings only represents the 24 schools included in the research, and cannot be generalized to other schools in Gauteng. Data used in this analysis formed part of the baseline assessment of the Skhokho RCT, thus other variables that are documented in the literature, and found to be associated with experience of CP such as neighbourhood and community level factors were not measured. Furthermore, the analysis was conducted from quantitative baseline data. A qualitative exploration of the topic might have provided further details about learner experiences of corporal punishment in schools. Given that use of CP is a behaviour of teachers, it is important to explore factors associated with teachers use of CP. Future qualitative research with learners and teachers is warranted to strengthen our understanding of not only the learners\u2019 experience of CP but also teachers\u2019 insights on why CP continues to be used at school, despite the legislation prohibiting its use.Our findings have shown that learners still experience CP in public schools in South Africa. While there is a need to also focus on learner behaviour, our findings have highlighted the need to intervene in both the home and school environments. There is an urgent need to break this cycle of violence, through use of CP, by enforcing the law, and holding accountable those who continue to use CP despite legislation prohibiting its use, but also supporting the children in their home environment, and their parents to positively partner with their children. Furthermore, there is a need to provide services to meet leaner\u2019s mental health needs. Evidence-based interventions are needed to support both parents and teachers in managing learner behaviour. Use of positive disciplining strategies, developing democratic relationship and consciousness about image of a child can positively informt how parents and teachers relate with children, critical for raising responsible children, and to curb future perpetration of violence in society , 56. Eff 1 Mar 2021PONE-D-20-20199Prevalence and factors associated with experience of corporal punishment in public schools in South AfricaPLOS ONEDear Dr. Mahlangu,Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE\u2019s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.plosone@plos.org. 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PLOS defines a competing interest as anything that interferes with, or could reasonably be perceived as interfering with, the full and objective presentation, peer review, editorial decision-making, or publication of research or non-research articles submitted to one of the journals. Competing interests can be financial or non-financial, professional, or personal. Competing interests can arise in relationship to an organization or another person. Please follow this link to our website for more details on competing interests: [Note: HTML markup is below. Please do not edit.]Reviewers' comments:Reviewer's Responses to QuestionsComments to the Author1. Is the manuscript technically sound, and do the data support the conclusions?The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1:\u00a0YesReviewer #2:\u00a0YesReviewer #3:\u00a0Yes**********2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1:\u00a0I Don't KnowReviewer #2:\u00a0I Don't KnowReviewer #3:\u00a0Yes**********3. Have the authors made all data underlying the findings in their manuscript fully available?PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data\u2014e.g. participant privacy or use of data from a third party\u2014those must be specified.The Reviewer #1:\u00a0YesReviewer #2:\u00a0YesReviewer #3:\u00a0Yes**********4. Is the manuscript presented in an intelligible fashion and written in standard English?PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.Reviewer #1:\u00a0YesReviewer #2:\u00a0YesReviewer #3:\u00a0Yes**********5. Review Comments to the AuthorPlease use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. Reviewer #1:\u00a0Important note: This review pertains only to \u2018statistical aspects\u2019 of the study and so \u2018clinical aspects\u2019  are to be evaluated [should be assessed] separately/independently. Further please note that any \u2018statistical review\u2019 is generally done under the assumption that (such) study specific methodological [as well as execution] issues are perfectly taken care of by the investigator(s). This review is not an exception to that and so does not cover clinical aspects {however, seldom comments are made only if those issues are intimately / scientifically related & intermingle with \u2018statistical aspects\u2019 of the study}. Agreed that \u2018statistical methods\u2019 are used as just tools here, however, they are vital part of methodology [and so should be given due importance].COMMENTS: Abstract-Methods section says \u201c3743 grade 8 learners (2118 girls and 1625 boys) from 24 selected public schools in Tshwane, South Africa, enrolled in a cluster randomised controlled trial\u201d my question is if the schools (clusters) are purposively selected, where is randomization? .Although, it is said in line 178 that [\u2018All data analysis procedures used took into account the study design\u2019], it is not observed. In this context , here are three good references.1. Donner Allen and Klar Neil. `Design and Analysis of Cluster Randomization Trial in Health Research\u2019, Oxford University Press Inc., New York, 2000.2. Bland JM and Kerry SM. \u2018Statistics notes: trials randomized in clusters\u2019, Br. Med. Jr., 1997, Volume 315, p600.3. Kerry SM & Bland JM. \u2018Analysis of a trial randomized in clusters\u2019 by in British Medical Journal, Volume 316, 1998, p54.Also note thatThough the measures/tools used are appropriate [Example - Line 112-3: A survey was conducted using a self-administered questionnaire loaded in personal digital assistants (PDAs)], most of them yield data that are in [at the most] \u2018ordinal\u2019 level of measurement [and not in ratio level of measurement for sure {as the score two times higher does not indicate presence of that parameter/phenomenon as double }]. Then application of suitable non-parametric test(s) is/are indicated/advisable [even if distribution may be \u2018Gaussian\u2019 ]. Agreed that there is/are no non-parametric test(s)/technique(s) available to be used as alternative in all situation(s) [suitable / most desired/applicable], but should be used wherever/wherever they are available.From Table 1 [Description of the factors that influence experience of corporal punishment at school] it appears that many measures used are of this type. I request the authors to also note that (this note is copied and pasted from a standard text book) : Whenever response options ranged from 1=strongly disagree to 4=strongly agree (or ranging from 1 (strongly disagree) to 6 (strongly agree) or from 1=very bad to 3=neither good nor bad to 5=very good), while using a \u2018Likert\u2019 scale responses, recoding  may yield correct and meaningful \u2018arithmetic mean\u2019 which is useful not only for comparison but has absolute meaning, in my opinion. Application of any statistical test(s) assume that meaning of entity used  has a particular meaning. Though \u2018\u03b1\u2019 [alpha] or most other measures of reliability/correlation will remain same, however. Use of non-parametric methods should/may be preferred while dealing with data yielded by any questionnaire/score.From account given in lines 228-9 [Table 3 shows the relationship between experiencing CP at school and individual measures of the different latent constructs] it is clear that in table-3 {Bivariate relationship between corporal punishment experience score and a learner\u2019s home environment, attitude and behavior measures.} first column Coef is \u2018Correlation Coefficient\u2019 (mostly Pearson\u2019s) {LCL is lower & UCL is upper limit of CI I guess, why not given in footnote). Coefficient values are very small, yet most p-values show highly significant results. Is not that contradictory? In this context again it may please be noted that (following note is from a standard text book):Statistical test usually used to assess significance of Pearson\u2019s \u2018Correlation coefficient (r)\u2019 is \u2018t\u2019 [where t = { r \ufffd [(n-2) / (1-r2)] }for df=n-2, n is sample size] and here Ho is that the population/standard value of \u2018r\u2019 is zero. You need r=0.878 to be significant at 5% when n=5 but you need r=0.273 if n=50 & you need only r=0.088 if n=500. \u2018P-value\u2019 heavily depends on sample size. Therefore, it is customary to use the (available in most text books on \u2018Biostatistics\u2019 or on \u2018www/net\u2019) guidelines [very strongly suggesting to consider an absolute value of \u2018Correlation coefficient\u2019] for interpreting positive or negative correlations . [This argument is equally applicable to non-parametric Spearman\u2019s \u2018Correlation coefficient (\u03c1)\u2019 as well.]Please look at an absolute value of \u2018Correlation coefficient\u2019 in table-3. It is suggested that \u2018If r = +.70 or higher Very strong positive relationship\u2019 or \u2018-.70 or higher Very strong negative relationship\u2019. In table-3, all \u2018Coef\u2019 are below 0.18 [smallest as small as 0.02]. It is not mentioned however that which type of correlation coefficient is used. But in case in the light of the above, interpretation given in lines 228 onwards is questionable.Moreover, as said in lines 140-145 CP at school is measured by score  and both \u2018simple linear regression\u2019 as well as \u2018correlation\u2019 {lines 181-3: We used simple linear regression to assess the relationship between experience of CP at school and all individual measures described in Table 1, and also assessed the correlation amongst the measures} assume that (both \u2018independent\u2019 & \u2018dependent\u2019) variables are continuous.Surprisingly use of \u2018structural equation modelling\u2019 seems to be correct. Account given in lines 184-198 seems perfectly alright. Figures 1,2,3 are alright. Look at the magnate of coefficients in contrast to coefficients reported/displayed in table-3, though they may be different [in nature & purpose].Reviewer #2:\u00a0This is an interesting quantitative study on the prevalence of corporal punishment amongst 8 graders in 24 selected government schools in South Africa. The study is trying to uncover the underlying reasons for its prevalence and suggests a solution to it.The article is well-written and makes a good contribution to the field in that it provides evidence for the fact that corporal punishment is happening. This is in itself important as research in CP is notoriously difficult, because CP is illegal since 1997 and could cause major problems for teachers if found out. It is interesting that this part of the study is not really discussed at all. Research ethics and what exactly was said to the school principals is missing. For a study of this kind it is important to also publish the letters of consent and assent and how some of that was negotiated. CP tends to go underground when there is a risk that teachers might get reported. But there is another issue worth mentioning and that is, if corporal punishment is defined as physical punishment, then why are the main questions of the survey phrased in terms of \u2018Were you beaten\u2026?\u2019. From my experiences of researching CP, beatings are a small part of the abuse that is routinely going on in schools. I would really like to see the full set of questions used in the study. The evidence sought was to identify the risk factors. There is interesting info about the characteristics of children who receive more CP in the literature, which is again confirmed by this study. A qualitative component probably would have given more unexpected results, because that is often \u2018hiding\u2019 in the details. I would very much like to have learned about teachers\u2019 perspectives, but how to conduct such a study (and get indeed genuine information) is another matter. However, I understand such a follow-up study is in the pipeline.The research was carried out by the SA Medical Research Council and it is within that context that some of my comments might be helpful. I am not an expert in quantitative studies and this one doesn\u2019t use a qualitative component , so I can\u2019t comment on the use of the methods that were used, but the following might be helpful:1. Line 26: To my knowledge CP is not prevalent in schools globally, maybe homes? It would have been helpful to be more specific as it is certainly not common in schools in Europe, Australia etc, although it might be normal in households as parents are often allowed to use CP but not teachers. The details are important here.2. Page 4: Factors that influence the use of CP in US are of use, but it would have been helpful to hear more about the racialised discourses that inform the use of CP - e.g. children are seen as wild and need to be tamed, domesticated and controlled. Innocence tends not to be attributed to brown bodies \u2013 a discourse also internalised by black people themselves. It was striking to see a lack of reference to racial, ethnic, religious and other such factors that are critical in relationships in SA. I would really like to see all research instruments included as Appendices.3. At times it isn't clear to which part of the world the info applies and claims are made. For example, first there is evidence from Pakistan, then later US, but on page 5 it is unclear whether the authors make claims about CP as a global phenomenon or not. The info needs to be tied more specifically to the geopolitical location. What is missing is background information about the racialised and historically complex two-tier education system. Classrooms are very large and on the whole teachers not as qualified as in many other places. Therefore, this article could unfairly maybe give a very negative impression of a terribly underpaid and under supported workforce. Teachers in SA work under very difficult conditions and this article should include more information about teachers\u2019 conditions so it doesn\u2019t run the risk of being accused of a deficit approach, especially as teachers\u2019 voice in the study is absent. It is only learners\u2019 perspectives.4. Line 90: This is a generalisation not supported by evidence. Education departments in SA Universities seem to be doing their upmost to teach their students alternative discipline strategies, but the phenomenon is more complex than that. Also Provincial education departments all have initiatives to support teachers in using different discipline methods, but the particular image of child and childhood and child/adult relations tend to be hierarchical in SA and lack the equality needed for respectful non-violent relations. This is typical of patriarchal societies. The violence against children in SA is intricately connected to the violence against women.5. What is completely missing is the relationship between the training of teachers, their ability to teach in ways that engages learners and motivates them. Teacher educators know that e.g. more active and embodied pedagogies can be used that prevent 'misbehaviour' from occurring. Large class sizes etc also really contribute to CP and not having classroom assistants.6. The intervention focuses on positive discipline and psychological interventions, but not on the image of the child or the pedagogical dimensions. So in a sense, the hypothesis of the study makes sense in the light of the identity of the researchers. It would be helpful to include in future research someone from a very different educational background so base-line hypotheses can be enriched. We know from educational research that teachers tend to teach how they were taught and it is very difficult to break through that despite the many interventions in teacher education and by provincial departments. Therefore, the suggestion that the solution to the problem identified might be to offer training in positive discipline is nothing new and has been going on for some time now. It is a good idea to check out this literature. What would be worthwhile and unusual and new instead is to include an in-service course/intervention in childhood studies (to teach about the image of the child) and the building of democratic relationships through e.g. \u2018shared authority\u2019 models. These are about more sustainable and durable changes as they could affect deeply engrained beliefs about children as found wanting (by adults), rather than interventions that don\u2019t change fundamental beliefs about people and their relations. However, this would require pedagogical skills and working with both groups at the same time: learners and their teachers.In short, this is a very informative article, well written and researched and helpful in that it confirms much of what we already know about CP, and indeed helpful for funding purposes, but it misses the important geopolitical educational dimension (which still can be added) and references to pedagogies which hopefully can be taken into account in further studies. I hope the above comments are helpful, in order to add further reflections to the current paper, but especially I hope they are useful for future research purposes.Reviewer #3:\u00a0This study examines the prevalence and factors associated with corporal punishment in public schools in South Africa using baseline data from a larger RCT study.This is a highly competent and well published team who have completed most of the seminal work on interpersonal violence in South Africa and in fact globallyThe methodology of the larger randomized controlled trial is excellent and it appears to be a well conducted studyI have one analytic concern. The authors merge two questions \u2018In the past 6 months were you ever beaten by a teacher?\u2019 and \u201cA teacher might beat you or physically punish you at our school\u201d. These are two entirely different questions and simply because most people answer yes to the first have answered yes to the second (of course they have) is no reason to merge them. This must be corrected in the revised version.The use of the term \u2018delinquent\u2019 is highly problematic, certainly not justified by the data collected in the paper and should be deleted. To describe a learner as delinquent based on 5 questions is highly problematic.A more general concern has to do with what these findings add to the literature. Corporal punishment is associated with learner behaviour, home environment and school environment (i.e. everything). It does feel a little like squeezing another paper out of the dataset. While the paper adds little to what is already known, the method and execution of the study are of a high standard and the findings may be of some interest to education authorities in South Africa.**********what does this mean?). If published, this will include your full peer review and any attached files.6. PLOS authors have the option to publish the peer review history of their article  digital diagnostic tool,\u00a0Attachmentrenamed_da594.docxSubmitted filename: Click here for additional data file. 3 May 2021Dear EditorThank you for the opportunity to revise our paper. Below we have described how we have responded to the comments from the reviewersBest regards,Reviewer 1 Important note: This review pertains only to \u2018statistical aspects\u2019 of the study and so \u2018clinical aspects\u2019  are to be evaluated [should be assessed] separately/independently. Further please note that any \u2018statistical review\u2019 is generally done under the assumption that (such) study specific methodological [as well as execution] issues are perfectly taken care of by the investigator(s). This review is not an exception to that and so does not cover clinical aspects {however, seldom comments are made only if those issues are intimately / scientifically related & intermingle with \u2018statistical aspects\u2019 of the study}. Agreed that \u2018statistical methods\u2019 are used as just tools here, however, they are vital part of methodology [and so should be given due importance]. Thank you very much for your time reviewing our manuscript and for the valuable feedback. We have revised and clarified the analytic methods applied to address issues raised, taking into account suggestions and recommendations from all the reviewers.Abstract-Methods section says \u201c3743 grade 8 learners (2118 girls and 1625 boys) from 24 selected public schools in Tshwane, South Africa, enrolled in a cluster randomised controlled trial\u201d my question is if the schools (clusters) are purposively selected, where is randomization? . While selection of the schools into the study was not random, the allocation of the selected schools to the interventions arms was random.Although, it is said in line 178 that [\u2018All data analysis procedures used took into account the study design\u2019], it is not observed. In this context , here are three good references.1. Donner Allen and Klar Neil. `Design and Analysis of Cluster Randomization Trial in Health Research\u2019, Oxford University Press Inc., New York, 2000.2. Bland JM and Kerry SM. \u2018Statistics notes: trials randomized in clusters\u2019, Br. Med. Jr., 1997, Volume 315, p600.3. Kerry SM & Bland JM. \u2018Analysis of a trial randomized in clusters\u2019 by in British Medical Journal, Volume 316, 1998, p54. We acknowledge the source of information provided on analysis of cluster randomized. In our analysis, we performed individual level analysis considering:i) We are analyzing baseline data for an outcome that is not primary trial outcome.ii) We have a sizable number of clusters per arm and we accounted for the clustering in the estimation of standard errors in all the models.Though the measures/tools used are appropriate [Example - Line 112-3: A survey was conducted using a self-administered questionnaire loaded in personal digital assistants (PDAs)], most of them yield data that are in [at the most] \u2018ordinal\u2019 level of measurement [and not in ratio level of measurement for sure {as the score two times higher does not indicate presence of that parameter/phenomenon as double ]. Then application of suitable non-parametric test(s) is/are indicated/advisable [even if distribution may be \u2018Gaussian\u2019 ]. Agreed that there is/are no non-parametric test(s)/technique(s) available to be used as alternative in all situation(s) [suitable / most desired/applicable], but should be used wherever/wherever they are available.From Table 1 [Description of the factors that influence experience of corporal punishment at school] it appears that many measures used are of this type. I request the authors to also note that (this note is copied and pasted from a standard text book) : Whenever response options ranged from 1=strongly disagree to 4=strongly agree (or ranging from 1 (strongly disagree) to 6 (strongly agree) or from 1=very bad to 3=neither good nor bad to 5=very good), while using a \u2018Likert\u2019 scale responses, recoding  may yield correct and meaningful \u2018arithmetic mean\u2019 which is useful not only for comparison but has absolute meaning, in my opinion. Application of any statistical test(s) assume that meaning of entity used  has a particular meaning. Though \u2018\u03b1\u2019 [alpha] or most other measures of reliability/correlation will remain same, however. Use of non-parametric methods should/may be preferred while dealing with data yielded by any questionnaire/score.From account given in lines 228-9 [Table 3 shows the relationship between experiencing CP at school and individual measures of the different latent constructs] it is clear that in table-3 {Bivariate relationship between corporal punishment experience score and a learner\u2019s home environment, attitude and behavior measures. first column Coef is \u2018Correlation Coefficient\u2019 (mostly Pearson\u2019s) {LCL is lower & UCL is upper limit of CI I guess, why not given in footnote). Coefficient values are very small, yet most p-values show highly significant results. Is not that contradictory? In this context again it may please be noted that (following note is from a standard text book):Statistical test usually used to assess significance of Pearson\u2019s \u2018Correlation coefficient (r)\u2019 is \u2018t\u2019 [where t = { r \ufffd [(n-2) / (1-r2)] }for df=n-2, n is sample size] and here Ho is that the population/standard value of \u2018r\u2019 is zero. You need r=0.878 to be significant at 5% when n=5 but you need r=0.273 if n=50 & you need only r=0.088 if n=500. \u2018P-value\u2019 heavily depends on sample size. Therefore, it is customary to use the (available in most text books on \u2018Biostatistics\u2019 or on \u2018www/net\u2019) guidelines [very strongly suggesting to consider an absolute value of \u2018Correlation coefficient\u2019] for interpreting positive or negative correlations . [This argument is equally applicable to non-parametric Spearman\u2019s \u2018Correlation coefficient (\u03c1)\u2019 as well.]Please look at an absolute value of \u2018Correlation coefficient\u2019 in table-3. It is suggested that \u2018If r = +.70 or higher Very strong positive relationship\u2019 or \u2018-.70 or higher Very strong negative relationship\u2019. In table-3, all \u2018Coef\u2019 are below 0.18 [smallest as small as 0.02]. It is not mentioned however that which type of correlation coefficient is used. But in case in the light of the above, interpretation given in lines 228 onwards is questionable. We acknowledge the concern raised regards the distributional assumptions for the analytical methods used in the bivariate analysis. We do agree that non-parametric tests are more robust than parametric test for discrete measures.Our aim for performing the correlations/linear regression analyses was mainly as an exploratory data analysis preceding the main analysis . This was done to show how the different measures were associated with the outcome , prior to the SEM analysis.Based on recommendation from the review, we have defined our outcome measure  as a binary outcome derived from responses from 1 question rather than from 2 questions as previously defined.We have thus revised Table 3 to reflect this change and we now present relationships using Odds ratios from logistic regression models.We have revised TablesMoreover, as said in lines 140-145 CP at school is measured by score  and both \u2018simple linear regression\u2019 as well as \u2018correlation\u2019 {lines 181-3: We used simple linear regression to assess the relationship between experience of CP at school and all individual measures described in Table 1, and also assessed the correlation amongst the measures} assume that (both \u2018independent\u2019 & \u2018dependent\u2019) variables are continuous. As indicated above, we have redefined the outcome using only 1 item (In the past 6 months were you ever beaten by a teacher?)Surprisingly use of \u2018structural equation modelling\u2019 seems to be correct. Account given in lines 184-198 seems perfectly alright. Figures 1,2,3 are alright. Look at the magnate of coefficients in contrast to coefficients reported/displayed in table-3, though they may be different [in nature & purpose]. Thank you. We have revised the SEM analysis and fitted a Generalised SEM with the experience of corporal punishment defined as a binary outcome.Reviewer #2 This is an interesting quantitative study on the prevalence of corporal punishment amongst 8 graders in 24 selected government schools in South Africa. The study is trying to uncover the underlying reasons for its prevalence and suggests a solution to it. Thank you very much for your time reviewing our manuscript and for the valuable feedback.The article is well-written and makes a good contribution to the field in that it provides evidence for the fact that corporal punishment is happening. This is in itself important as research in CP is notoriously difficult, because CP is illegal since 1997 and could cause major problems for teachers if found out. It is interesting that this part of the study is not really discussed at all. Research ethics and what exactly was said to the school principals is missing. For a study of this kind it is important to also publish the letters of consent and assent and how some of that was negotiated. CP tends to go underground when there is a risk that teachers might get reported. But there is another issue worth mentioning and that is, if corporal punishment is defined as physical punishment, then why are the main questions of the survey phrased in terms of \u2018Were you beaten\u2026?\u2019. From my experiences of researching CP, beatings are a small part of the abuse that is routinely going on in schools. I would really like to see the full set of questions used in the study. The evidence sought was to identify the risk factors. There is interesting info about the characteristics of children who receive more CP in the literature, which is again confirmed by this study. A qualitative component probably would have given more unexpected results, because that is often \u2018hiding\u2019 in the details. I would very much like to have learned about teachers\u2019 perspectives, but how to conduct such a study (and get indeed genuine information) is another matter. However, I understand such a follow-up study is in the pipeline. a. The use of corporal punishment in schools is prohibited by law in South Africa. Any person who administers corporal punishment at school is guilty of an offence and liable on conviction to a sentence which could be imposed for assault. This has been added in the introduction section on page:The South African Schools Act No 84 of 1996 states that no person may administer CP to a learner at school (1). Any person who administers corporal punishment at school is guilty of an offence and liable on conviction to a sentence which could be imposed for assault (1). b. We have added on page 9 line 138 a sub-heading \u2018Ethical considerations. We have also added a paragraph in line 141 \u2013 148 explaining what was said to principals when requesting permission to work in schools:Permission was also granted by the school principals in the 24 schools. The information letter detailing the nature and purpose of the study, the interventions, participants, what the project would offer to schools, risks and benefits of participating in the study, project timelines, and on rights of participants was provided to principals. School principals were also informed that all data generated from the study will be kept confidential and that research reports and articles which will be submitted in scientific journals will not include any information that may identify the school or any of the educators, learners, school governing body officials and parents. c. We agree with the reviewer that CP tends to go underground, underreported when there is a risk that teachers might get reported. This is also shown in studies where both learners and teachers have been involved and the findings showed varied reports of use of corporal punishment in schools, learners reporting higher prevalence that teachers. In our study, use of corporal punishment in the 24 schools was reported by learnersd. Table 1 describes the main aspects of the questionnaire e. We used the Childhood Trauma Questionnaire and cognitively tested the questionnaire amongst grade 8 learners in schools which were not part of the research and found that \u2018beaten\u2019 was understood and captured corporal punishment as was intendedf. We have expanded the limitations section on page 27, line 439 - 445 to acknowledge the limitation of not including a qualitative component and teacher\u2019s perspectives:Furthermore, the analysis was conducted from quantitative baseline data. A qualitative exploration of the topic might have provided further details about learner experiences of corporal punishment in schools. Given that use of CP is a behaviour of teachers, it is important to explore factors associated with teachers use of CP. Future qualitative research with learners and teachers is warranted to strengthen our understanding of not only the learners\u2019 experience of CP but also teachers\u2019 insights on why CP continues to be used at school, despite the legislation prohibiting its use. The research was carried out by the SA Medical Research Council and it is within that context that some of my comments might be helpful. I am not an expert in quantitative studies and this one doesn\u2019t use a qualitative component , so I can\u2019t comment on the use of the methods that were used, but the following might be helpful: Thank you very much for your time reviewing our manuscript and for the valuable feedback1. Line 26: To my knowledge CP is not prevalent in schools globally, maybe homes? It would have been helpful to be more specific as it is certainly not common in schools in Europe, Australia etc, although it might be normal in households as parents are often allowed to use CP but not teachers. The details are important here. While use of corporal punishment in school is banned and no longer an issue in European countries, it is a problem that requires attention in many other parts of the world. Corporal punishment is still prevalent in a third of the world\u2019s countries where it continues to be used as a method of disciplining children, both in countries where it is legal and countries where it is banned, leading to estimates that millions of children are subject to this harmful practice . We have revised the introduction section to become more specific about prevalence of CP globally, line 25 \u2013 31:While the use of a physical methods of discipline, also referred to as corporal punishment (CP) at school is banned and no longer an issue in European countries; it continues to be prevalent in a third of the world\u2019s countries, despite evidence regarding its harmful physical, mental and behavioural effects on the child . The global prevalence of CP in schools ranges between 13% \u2013 97% of learners who reported experience of CP at school (1). Learners continue to experience CP in most countries in Sub-Saharan Africa despite legislation prohibiting its use .2. Page 4: Factors that influence the use of CP in US are of use, but it would have been helpful to hear more about the racialised discourses that inform the use of CP - e.g. children are seen as wild and need to be tamed, domesticated and controlled. Innocence tends not to be attributed to brown bodies \u2013 a discourse also internalised by black people themselves. It was striking to see a lack of reference to racial, ethnic, religious and other such factors that are critical in relationships in SA. I would really like to see all research instruments included as Appendices. We agree with the reviewer that race, ethnicity and religion are critical in relationships in South Africa. However, unlike in American schools, most public schools in South Africa are still very much racially segregated by communities served in that area. While the system of schools divided by race was eliminated in 1996s, public schools in Black townships or communities are racially homogenous, while the historically White and Indian schools serve a more racially diverse constituency . About 21 of the schools where this research was conducted were black only schools: Only three of the 24 schools were mixed race schools. Even in the three mixed race schools, more than 90% of the students were black and this is shown in Table 2 on page 16The reviewer is correct about the diversity in religion and ethnicity in schools in South Africa, recognized in the National Policy on Religion and Education of 2003. It is possible that religion and ethnicity could have been amongst the factors associated with learner experience of corporal punishment in schools, but we did not include in our analysis. 3. At times it isn't clear to which part of the world the info applies and claims are made. For example, first there is evidence from Pakistan, then later US, but on page 5 it is unclear whether the authors make claims about CP as a global phenomenon or not. The info needs to be tied more specifically to the geopolitical location. What is missing is background information about the racialised and historically complex two-tier education system. Classrooms are very large and on the whole teachers not as qualified as in many other places. Therefore, this article could unfairly maybe give a very negative impression of a terribly underpaid and under supported workforce. Teachers in SA work under very difficult conditions and this article should include more information about teachers\u2019 conditions so it doesn\u2019t run the risk of being accused of a deficit approach, especially as teachers\u2019 voice in the study is absent. It is only learners\u2019 perspectives. We have revised the introduction to include background information about the racialized and historically complex two-tier education system in South Africa in line 34 - 38: Corporal punishment has been an integral part of schooling for most teachers and learners in twentieth century South African schools, characterized by a legacy of authoritarian education practices under Bantu education and a belief that CP is necessary for orderly education (7). The ending of apartheid and the establishment of a human rights culture in the 1990s laid the foundation for legislation aimed at ending use of CP in schools in South Africa (7). Line 40 \u2013 44:South Africa is a highly inequitable society and this inequality is reflected in the historically complex two-tier education system namely, the middle class, private schools and the public schools (5). The middle class, formerly white schools no longer use corporal punishment as a discipline method (7). However, in public schools, use of corporal punishment is still common practice We agree with the reviewer that teachers, particularly in public schools, work under difficult conditions and this background information has now been added in line 44 \u2013 47:In most public schools, classrooms are overcrowded and under-resourced, and teachers are often under-qualified and overworked . Teachers feel disempowered and ill-equipped with viable alternative discipline methods to maintain a safe and secure environment to facilitate learning (8)4. Line 90: This is a generalisation not supported by evidence. Education departments in SA Universities seem to be doing their upmost to teach their students alternative discipline strategies, but the phenomenon is more complex than that. Also Provincial education departments all have initiatives to support teachers in using different discipline methods, but the particular image of child and childhood and child/adult relations tend to be hierarchical in SA and lack the equality needed for respectful non-violent relations. This is typical of patriarchal societies. The violence against children in SA is intricately connected to the violence against women. We have revised the paragraph. It now reads as follows in line 109 - 112:While use of CP is illegal in South Africa, there has been limited concerted effort to enforce the law, ensuring that those who continue to use CP are convicted of an offense, and inadequate training of teachers on alternative methods of classroom management and discipline More can be done to enforce implementation of legislation and holding accountable those who illegally continue to use CP despite legislation prohibiting its use. Gershoff points that: legal bans are not sufficient to completely eliminate school corporal punishment. Behavior change by teachers and school administrators is what is needed. Behaviour change will require consistent education about the harms of corporal punishment and about alternative, positive forms of discipline. While there might be pockets of schools where teachers are exposed to initiatives aimed to use positive discipline methods, not all schools have been provided with such, and the support has not been consistent. We have revised the conclusion, line 453 \u2013 455:There is an urgent need to break this cycle by enforcing the law, and holding accountable those who continue to use CP despite legislation prohibiting its useAnd line 459 \u2013 464:Use of positive disciplining strategies, developing democratic relationship and consciousness about image of a child will direct how parents and teachers relate with children, critical for raising responsible children, and to curb future perpetration of violence in society  Effecting engrained beliefs, attitudes and behaviours supportive of corporal punishment amongst parents and teachers will require consistent education about the harms of corporal punishment and support for why it needs to be ended. 5. What is completely missing is the relationship between the training of teachers, their ability to teach in ways that engages learners and motivates them. Teacher educators know that e.g. more active and embodied pedagogies can be used that prevent 'misbehaviour' from occurring. Large class sizes etc also really contribute to CP and not having classroom assistants. We agree with the reviewer that there are many factors that could explain use of corporal punishment amongst teachers including factors related to their training. It is likely that these factors are contributing to their ability/ inability to teach in ways that engages the learner and motivates them, so as to manage some of the risk factors associated with learner experience of CP. As suggested by the reviewer, a study which captures teacher\u2019s perspectives on use of corporal punishment in schools will contribute to knowledge and respond to these questions. We have not included teachers\u2019 perspectives on this paper, and plan to do so in future. 6. The intervention focuses on positive discipline and psychological interventions, but not on the image of the child or the pedagogical dimensions. So in a sense, the hypothesis of the study makes sense in the light of the identity of the researchers. It would be helpful to include in future research someone from a very different educational background so base-line hypotheses can be enriched. We know from educational research that teachers tend to teach how they were taught and it is very difficult to break through that despite the many interventions in teacher education and by provincial departments. Therefore, the suggestion that the solution to the problem identified might be to offer training in positive discipline is nothing new and has been going on for some time now. It is a good idea to check out this literature. What would be worthwhile and unusual and new instead is to include an in-service course/intervention in childhood studies (to teach about the image of the child) and the building of democratic relationships through e.g. \u2018shared authority\u2019 models. These are about more sustainable and durable changes as they could affect deeply engrained beliefs about children as found wanting (by adults), rather than interventions that don\u2019t change fundamental beliefs about people and their relations. However, this would require pedagogical skills and working with both groups at the same time: learners and their teachers. We thank the reviewer for this comment, which we have noted. We will ensure that in future we involve someone with education background to help enrich our work by incorporating a focus on the image of the child or the pedagogical dimensions.We have incorporated the reviewer\u2019s suggestions on strategies to address corporal punishment in schools, line 459 \u2013 464:Use of positive disciplining strategies, developing democratic relationship and consciousness about image of a child will direct how parents and teachers relate with children, critical for raising responsible children, and to curb future perpetration of violence in society  Effecting engrained beliefs, attitudes and behaviours supportive of corporal punishment amongst parents and teachers will require consistent education about the harms of corporal punishment and support for why it needs to be ended. In short, this is a very informative article, well written and researched and helpful in that it confirms much of what we already know about CP, and indeed helpful for funding purposes, but it misses the important geopolitical educational dimension (which still can be added) and references to pedagogies which hopefully can be taken into account in further studies. I hope the above comments are helpful, in order to add further reflections to the current paper, but especially I hope they are useful for future research purposes. We have revised the paper and included recommendations on teaching practices that are critical in efforts to end use of corporal punishment in schools, line 459 \u2013 464:Use of positive disciplining strategies, developing democratic relationship and consciousness about image of a child will direct how parents and teachers relate with children, critical for raising responsible children, and to curb future perpetration of violence in society  Effecting engrained beliefs, attitudes and behaviours supportive of corporal punishment amongst parents and teachers will require consistent education about the harms of corporal punishment and support for why it needs to be ended. Reviewer 3 This study examines the prevalence and factors associated with corporal punishment in public schools in South Africa using baseline data from a larger RCT study.This is a highly competent and well published team who have completed most of the seminal work on interpersonal violence in South Africa and in fact globallyThe methodology of the larger randomized controlled trial is excellent and it appears to be a well conducted study Thank you very much for your time reviewing our manuscript and for the valuable feedback.I have one analytic concern. The authors merge two questions \u2018In the past 6 months were you ever beaten by a teacher?\u2019 and \u201cA teacher might beat you or physically punish you at our school\u201d. These are two entirely different questions and simply because most people answer yes to the first have answered yes to the second (of course they have) is no reason to merge them. This must be corrected in the revised version. Thank you for highlighting this point. We have corrected this. We have redefined the outcome using only 1 item (In the past 6 months were you ever beaten by a teacher?)The use of the term \u2018delinquent\u2019 is highly problematic, certainly not justified by the data collected in the paper and should be deleted. To describe a learner as delinquent based on 5 questions is highly problematic. We have replaced the term delinquent with misbehaviour throughout the manuscriptA more general concern has to do with what these findings add to the literature. Corporal punishment is associated with learner behaviour, home environment and school environment (i.e. everything). It does feel a little like squeezing another paper out of the dataset. While the paper adds little to what is already known, the method and execution of the study are of a high standard and the findings may be of some interest to education authorities in South Africa. The revisions based on the comments from the reviewers have improved the paper. While a lot has been written about corporal punishment in schools, very few published papers discuss the prevalence and use SEM to describe the interrelationships between factors associated with learner experience of CP in schools in South Africa. Like in other countries, South African schools are contextually unique, and deserving to be studied so we can have a better understanding of factors associated with use of CP in schools in South Africa. Given that CP is prohibited, yet continues to be used says that there is a problem, which needs to be engaged with, and recommended made on how it can be addressed. As such, this paper contributes to literature on the prevalence and factors associated with learners experiences of corporal punishment in schools in South Africa, and makes recommendation on what can be done to curb the use of corporal punishment in schools.References 1. Republic of South Africa. South African Schools Act No 84 of 1996. In: Office Ps, editor. Pretoria, South Africa: Government Printers; 1996. p. 1 - 50.2. Gershoff ET. School corporal punishment in global perspective: prevalence, outcomes, and efforts at intervention. Psychology, health & medicine. 2017;22(sup1):224-39.3. Heekes S-L, Kruger CB, Lester SN, Ward CL. A systematic review of corporal punishment in schools: Global prevalence and correlates. Trauma, Violence, & Abuse. 2020:1524838020925787.4. Munir A, Hussain B. Implications of Corporal Punishment on the Child's Mental Health in Peshawar, Pakistan. Pakistan Journal of Criminology. 2019;11(1).5. Spaull N. Poverty & privilege: Primary school inequality in South Africa. International Journal of Educational Development. 2013;33(5):436-47.6. Staeheli LA, Hammett D. \u2018For the future of the nation\u2019: Citizenship, nation, and education in South Africa. Political Geography. 2013;32:32-41.7. Morrell R. Corporal punishment in South African schools: A neglected explanation for its existence. South African Journal of Education. 2001;21(4):292-9.8. Maphosa C, Shumba A. Educators' disciplinary capabilities after the banning of corporal punishment in South African schools. South African Journal of Education. 2010;30(3):0-.9. Breen A, Daniels K, Tomlinson M. Children's experiences of corporal punishment: a qualitative study in an urban township of South Africa. Child abuse & neglect. 2015;48:131-9.10. Phurutse MC. Factors affecting teaching and learning in South African public schools: HSRC press; 2005.11. Mji A, Makgato M. Factors associated with high school learners' poor performance: a spotlight on mathematics and physical science. South African journal of education. 2006;26(2):253-66.12. Marumo M, Zulu C. Teachers\u2019 and learners\u2019 perceptions of alternatives to corporal punishment: A human rights perspective. In: Zulu C, Oosthuizen I, Wolhuter C, editors. A scholarly inquiry into disciplinary practices in educational institutions. 2. Cape Town, South Africa: AOSIS; 2019. p. 45 - 66.AttachmentRebuttal letter.docxSubmitted filename: Click here for additional data file. 28 Jun 2021Prevalence and factors associated with experience of corporal punishment in public schools in South AfricaPONE-D-20-20199R1Dear Dr. Mahlangu,We\u2019re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.Within one week, you\u2019ll receive an e-mail detailing the required amendments. When these have been addressed, you\u2019ll receive a formal acceptance letter and your manuscript will be scheduled for publication.http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at onepress@plos.org.If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they\u2019ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact Kind regards,Thach Duc Tran, M.Sc., Ph.D.Academic EditorPLOS ONEAdditional Editor Comments :Reviewers' comments:Reviewer's Responses to QuestionsComments to the Author1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the \u201cComments to the Author\u201d section, enter your conflict of interest statement in the \u201cConfidential to Editor\u201d section, and submit your \"Accept\" recommendation.Reviewer #1:\u00a0All comments have been addressedReviewer #2:\u00a0All comments have been addressed**********2. Is the manuscript technically sound, and do the data support the conclusions?The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1:\u00a0YesReviewer #2:\u00a0Yes**********3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1:\u00a0YesReviewer #2:\u00a0I Don't Know**********4. Have the authors made all data underlying the findings in their manuscript fully available?PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data\u2014e.g. participant privacy or use of data from a third party\u2014those must be specified.The Reviewer #1:\u00a0YesReviewer #2:\u00a0Yes**********5. Is the manuscript presented in an intelligible fashion and written in standard English?PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.Reviewer #1:\u00a0YesReviewer #2:\u00a0Yes**********6. Review Comments to the AuthorPlease use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. Reviewer #1:\u00a0COMMENTS: Most of the comments made on earlier draft {by me and hopefully by other respected reviewers also) were/are attended adequately. The manuscript is improved a lot. May be accepted if found clinically relevant.https://www.un.org/development/desa/family/wp-content/uploads/sites/23/2020/06/Freer_Expert-Group-Paper_Corporal-Punishment-Physical-Abuse_June2020.pdfReviewer #2:\u00a0Thank you for addressing my concerns quite meticulously. One of my key concerns does remain though and that is the narrow way in which CP has been defined in the study. Of course that can no longer be retified as it has been asumed from the outset and informed tha data collection. Corporal punishment is broader than 'being beaten'. Research has thrown up how much pain and physical discomfort is caused without beating a child. This can be by letting children stand for hours in a hot sun with their hands above their heads and without water or let them stand in a particular position against the wall for a long time so their legs will start to hurt. There are many ways in which children are hurt daily without being beaten. That is also corporal punishment. See eg the following link for the compexity in defining CP. This needs to be better explored and addressed in future studies. **********what does this mean?). If published, this will include your full peer review and any attached files.7. PLOS authors have the option to publish the peer review history of their article (If you choose \u201cno\u201d, your identity will remain anonymous but your review may still be made public.Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.Yes:\u00a0Dr. Sanjeev SarmukaddamReviewer #1:\u00a0Yes:\u00a0Professor Karin MurrisReviewer #2:\u00a0 4 Aug 2021PONE-D-20-20199R1 Prevalence and factors associated with experience of corporal punishment in public schools in South Africa. Dear Dr. Mahlangu:I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. onepress@plos.org.If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact plosone@plos.org. If we can help with anything else, please email us at Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staffon behalf ofDr. Thach Duc Tran Academic EditorPLOS ONE"}
+{"text": "During the COVID-19 pandemic, most countries took precautionary steps to save their citizens by initiating a lockdown and stopping all social activities by closing schools, companies, entertainment places, markets, gardens, and other social gathering places. As children stayed at home with no physical activities, their weight may have increased. The purpose of this study was to examine the link between fast food, sugars, or soft drinks and the ongoing domestic lockdown of the COVID-19 pandemic. This phenomenon was studied in three different cities from three different countries  from the perspective of children\u2019s parents. The study sought to address three research questions regarding children\u2019s well-being during the COVID-19 lockdown period. First, was children\u2019s nutrition affected during this period? Second, did children's weight increase? Third, were there any statistically significant differences in children\u2019s dietary patterns based on their gender and nationality? A questionnaire was administered to 330 parents of children aged four to seven years in the three targeted countries. The study found that most parents cared about their children's nutrition and prepared food at home (96.1%) during the lockdown. Sixty-three percent of parents indicated that children did not gain weight. Additionally, differences in children\u2019s nutritional systems were found between Saudi and Turkish children; the nutritional system of the Turkish children was better than that of Saudi children during the lockdown. Additionally, there were statistically significant differences in children\u2019s nutrition due to gender, with better nutrition for boys than for girls. On March 11, 2020, the World Health Organization announced that the novel COVID-19 virus had become a global pandemic. The rapid spread of the virus has resulted in some urgent temporary measures such as lockdowns in schools and universities all over the world. Most countries locked down educational institutions until further notice, with the provision of appropriate alternatives, such as the activation of\u00a0\u201cvirtual schools\u201d and distance learning classes.Children and adolescents' lifestyle behaviors, such as their physical activity and nutrition, may have been drastically impacted by prolonged school closures and home confinement during the COVID-19 pandemic. During the pandemic, the concept of self-care has grown. According to the National Alliance of Mental Health, the concept of self-care is a decision you take to improve your health, and it should be practiced daily. It includes body movement, health, nutrition, sleep, rest, and physical touch. The family or the children\u2019s caregivers should educate children about health care during this period, especially eating behaviors. These behaviors include eating healthy food and adhering to healthy habits such as personal hygiene. Moreover, the partnership between UNICEF and the World Food Program concentrated on intensive programs to protect, promote, and support optimal nutrition practices recommended for infants and young children, while adhering to health precautions, including stay-at-home procedures UNICEF, . Most goIn addition, with the spread of COVID-19, children are obliged to stay home for a long period of time because of the lockdown. Social media and magazines in Saudi Arabia focused on how the lockdown was causing people to gain weight . Staying home for a long period of time might affect children\u2019s weight. Young children are unable to choose the food they eat, just as they are unable to predict the long-term effects of their nutritional behavior; therefore, children need support and education in this regard  of the United Nations argued that when schools are closed or suspended, parents, caregivers, and children should be educated and informed about the importance of having safe, healthy meals. In addition, the FAO highlighted the importance of personal hygiene and physical exercise for school-age children. During the COVID-19 pandemic, some researchers conducted studies that addressed the issue of children who live in poor regions, and the extent of their vulnerability to this pandemic. On the other hand, few studies dealt with the health habits of children during the stay-at-home period. Research has also been conducted to link staying at home and gaining weight, obesity, and eating unhealthy food  during the COVID-19 lockdown from the perspectives of their parents. This study was approved by the local ethical review board and authorized by the Ministry of Education. In addition, to obtain parental permission, it was advised that a parent should read and go through the survey beforehand, and once they approved their child\u2019s participation, they read it to their child and recorded their answers. All subjects included in this research provided written informed consent before participation, and all data were used for this research and kept confidential. The principal investigator sent the IRB approval\u00a0with the survey to teachers and academic members in the UK and Turkey. The survey was distributed via Google Forms (because of the restrictions due to the pandemic) to different schools along with families and friends, targeting boys and girls aged four to seven years.The survey used was designed and reviewed by a panel of experts, comprised of a jury of faculty members at the Education Faculty of [blinded for review]. A survey was prepared as it was deemed the most appropriate means of collecting the data necessary to prove or reject the study hypotheses. The survey questions were designed according to the study goals and hypotheses, and based on cumulative knowledge obtained from theoretical readings within the framework of the subject of the study that contained a set of phrases and response choices. The respondents were asked to answer according to what they deemed appropriate for themselves and applied to their reality.The survey was sent to a sample of Saudi, Turkish, and British families through a Google Form randomly, containing a series of yes and no questions. To ensure the accuracy of the survey phrases in measurement variables, the validity of the tool was verified by a group of specialized faculty members and specialists in the College of Education, Department of Early Childhood at King Saud University. Some adjustments were made to the tool, in formulating some vocabulary, removing other vocabulary, and adding new vocabulary. After confirming the validity of the scale, Cronbach's alpha was calculated as a measure of the stability of the respondents' answers. The internal consistency of the study sample was 0.82, which indicates a high degree of consistency and can be relied upon in the field based on the reliability of its results. Internal reliability, which measures the degree of correlation between different items of the questionnaire, was assessed with a Pearson's correlation coefficient for knowledge scores, and a statistical significance level (0.01). Validity was assessed by Cronbach's alpha (0.65), which indicates that the study instrument has acceptable stability in the field application of the study.The randomly-chosen research sample consisted of parents of 330 children from the studied countries as follows: Saudi Arabia (198), British (62), and Turkish (70). Most of the study sample was\u00a0comprised of\u00a0Saudi children (60.0%), while the percentage of Turkish children was 21.2%, and the percentage of British children was 18.8%. Mothers comprised the majority of the study sample: there were 313 mothers (94.8%) and 17 fathers (18.8%). The majority of the children were seven years old , with four-year-olds accounting for 26.4% (87 children), five-year-olds for 19.7% (65 children), and six-year-olds for 17.0% (56 children). The total number of male children participating in the study was 160 (48.5%), whereas the total number of female children participating was 170 (51.5%).Data collected were analyzed using SPSS software. Pearson\u2019s correlation was used to assess the external construct validity, in which a statistical function belongs at the significance level (0.01) of all the axes of the items. In addition, the factors showed acceptable levels of internal reliability and consistency . Cronbach\u2019s alpha coefficients were computed to determine the reliability of each factor. The data were analyzed by frequency, percentage, and chi-square since the variables in question included three (3) attributes for \u201cyes\u201d,\u00a0 \u201csometimes\u201d, and \u201cno\u201d. Demographic related characteristics including age and sex were analyzed using frequencies, percentages, means, and standard deviations. In addition, the independent samples t-test provided information about whether the distributions of scores in the two independent groups were drawn from two identical population distributions , 50.6% responded \u201cno,\u201d which means that increases in children\u2019s weight were not due to unhealthy food and may be attributed to other factors such as the limited activities that they can engage in at home, the long hours of using digital games, and the unstable system of sleeping (children tend to play more and sleep less at inconsistent times). Finally, the parents' responses to item 11 also revealed a convergence in the responses of the parents on whether cake, biscuits, and cupcakes are unhealthy or healthy food: 41.2% responded \u201cno\u201d while 35.5% responded \u201cyes.\u201dIn addition, 50.5% of the responses of the Saudi participants to the direct question about the child\u2019s weight increase during the lockdown period were \u201cno.\u201d This high rate raises a concern regarding the causes of weight increase among Saudi children, despite the fact that food was home-prepared, and quick meal-ordering was limited. The Turkish response in this regard was 44.3% \u201cno,\u201d and the British response was 58.1%. To the question of \u201cUnhealthy food increased during the lockdown,\u201d 72.9% of Turkish participants and 61.3% of the British participants answered \u201cno.\u201dThe second question is whether there are any statistically significant differences in child nutrition during the COVID-19 pandemic lockdown in Saudi Arabia, Turkey, and the United Kingdom, attributable to nationality. To determine the effect of the child\u2019s nutritional system during this period, an ANOVA analysis was conducted. The results suggested significant differences, as shown in Table Table It is clear from Table The third question was whether there were any statistically significant differences in child nutrition attributed to gender during the COVID-19 pandemic stay-at-home period in Saudi Arabia, the United Kingdom, and Turkey. To answer this question, an independent samples test t-test was conducted. The results illustrate the significance of the differences in child nutrition during the COVID-19 pandemic lockdown that are attributed to gender. Table With the spread of COVID-19, social media and magazines in Saudi Arabia are discussing the lockdown and how people are gaining weight. Researchers anticipate that many children may experience higher-calorie diets during the pandemic response due to the COVID-19 lockdown ; but fewer quantities of fish (17.9%). Parents need to know the basic principles of meal preparation for children and the portions of different types of proteins children require in their meals , while Saudi children scored (44.4%), the weight gain of Turkish children was (32.9%), while Saudi children scored (33.3%), which is convergent. Moreover, 72.9% of the Turkish participants responded \u201cno\u201d to the question of whether there had been an increase in unhealthy food consumption during the lockdown period, versus 60.1% of the Saudi participants.\u201d This confirms that Turkish families value the importance of healthy meals and confirms the findings of Ayranci et al. (2009).Healthy food habits and behaviors include drinking water, eating vegetables and fruits, and for some individuals drinking milk might also be considered a healthy food habit. Turkish children scored higher than Saudi and British children. This calls for further research to examine Turkish nutritional habits, which could educate Saudi and British communities about young children\u2019s healthy nutritional systems.When parents in the three studied countries were asked whether their children were having cookies, cupcakes, or biscuits, 36.9% of parents in Saudi Arabia responded \u201cyes.\u201d The other two countries were convergent and parents responded with \u201cno\u201d . This raises questions about whether Saudi Arabian parents consider this type of food to be healthy.This study also concluded that there were statistically significant differences in favor of boys regarding eating healthy foods, and this invites parents to focus on their daughters' nutrition as they grow rapidly during this important stage in their lives ."}
+{"text": "Escherichia coli, Deinococcus radiodurans and Paraburkholderia fungorum under Mars-simulated conditions for freeze-thawing (\u221280 \u00b0C to +30 \u00b0C) and UV exposure alone and in combination. E. coli ATCC 25922, D. radiodurans and P. fungorum remained viable following 20 successive freeze-thaw cycles, exhibiting viabilities of 2.3%, 96% and 72.6%, respectively. E. coli ATCC 9079 was non-recoverable by cycle 9. When exposed to UV irradiation, cells withstood doses of 870 J/m2 (E. coli ATCC 25922), 200 J/m2 (E. coli ATCC 9079), 50,760 J/m2 (D. radiodurans) and 44,415 J/m2 (P. fungorum). Data suggests P. fungorum is highly UV-resistant. Combined freeze-thawing with UV irradiation showed freezing increased UV resistance in E. coli ATCC 25922, E. coli DSM 9079 and D. radiodurans by 6-fold, 30-fold and 1.2-fold, respectively. Conversely, freezing caused P. fungorum to exhibit a 1.75-fold increase in UV susceptibility. Strain-dependent experimentation demonstrated that freezing increases UV resistance and prolongs survival. These findings suggest that exposure to short wavelength UV rays (254 nm) and temperature cycles resembling the daily fluctuating conditions on Mars do not significantly affect survival of D. radiodurans, P. fungorum and E. coli ATCC 25922 following 20 days of exposure.The purpose of this study was to determine survivability of  Mars is a dry and barren planet with promising potential for future human colonization. Over the course of one Martian day 24.65 h), surface temperatures on Mars can vary depending on the location and the reporting source ,2. At th4.65 h, sMuch of the Martian surface is covered in a deep layer of dust with a primary composition of iron(III) oxide . MartianThe colonization of Mars has been traditionally mapped out via a terraforming approach. The terraforming of Mars is a multi-faceted operation, which, according to Haynes and McKay, involves three main steps; the human/robotic exploration of Mars (which has been conducted through various Mars landers), the planetary engineering step; designed to warm the planet, liberate liquid water and produce a thick carbon dioxide atmosphere; and the introduction of pioneering microbial communities able to proliferate in the Martian environment . While tIn order for bioremedial microorganisms to be successfully applied within these environments, it is imperative to understand how certain candidate microbes behave under the insult of Mars-like conditions. While the generation of terrestrial atmospheric conditions can be easily established on Mars, the immediate, and most threatening environmental insults to microbial life are daily temperature fluctuation (on average \u221263 \u00b0C to +30 \u00b0C) and UV radiation (UVC (200\u2013280 nm) and UVB (280\u2013315 nm)). Temperature cycling aims to recreate and determine the effects of daily Martian temperature fluctuations on microbial life. Similarly, in order to analyze the effects of shorter wavelengths  on the viability of the candidate microbes, dose-response experimentation was carried out to determine the limits of survival of the candidate strains in response to Mars-like UVC exposure.Escherichia coli ATCC 25922 and Escherichia coli DSM 9079 are both well-characterized microbes and functioned as the biological controls for both the freeze-thaw and UVC testing within the study. E. coli ATCC 25922 contains a functioning RecA gene, while E. coli DSM 9079 is a \u0394recA mutant that exhibits significant sensitivity to DNA damaging agents  = Power per m2 [W/m2] \u00d7 Exposure Time [s]. Power was determined by dividing the measured wattage reading from the UV energy detector by the energy detectors surface area (0.7 cm2). The CAMAG Cross-linker sample exposure area was mapped prior to experimentation using the optical power meter to determine the UV energy focal point. This area was marked, and test samples were positioned at this point to ensure consistency. This was performed for each experimental repeat to ensure an equal dose was delivered for all replicates. Prior to use, the CAMAG Cross-linker was sterilized with 70% Isopropyl-alcohol. A 0.9% saline solution was used to standardize prepared cells to a concentration of an OD600 nm of 1.0. A 1 mL volume of the biological replicate test samples  was placed into sterile mini-tissue-culture plates  and swirled gently to ensure an even surface distribution. Samples were irradiated singly. For samples with long exposure times, the sample dishes were swirled at 10 min intervals and 45 \u00b5L of 0.9% saline was added to account for the loss in sample volume due to evaporation . Once the desired dose of UVC was delivered, the samples were mixed gently, and 1 mL was placed into a sterile 1.5 mL tube. A volume of 100 \u03bcL was taken from the tube and plated onto sterile nutrient agar plates using the standard spread plate method. A limited number of low-dose-exposure samples required 1:10 serial dilutions using sterile Ringers solution, to aid in enumeration, when the colony forming unit (CFU) counts were too numerous to count. Once the sample had been exposed to the first dose of energy and plated, a further 1 mL volume of sample was added to a new mini-tissue-culture plate  and exposed to a greater dose of energy until completed and plated as described. This was repeated until each dose of energy had been administered. Both E. coli strains were incubated at 37 \u00b0C for 18 h, while P. fungorum and D. radiodurans were incubated at 30 \u00b0C for 48 h; all were then enumerated. It must be noted that only one replicate was prepared and irradiated at a time. See A CAMAG Cross-linker, housing a 254 nm 8 Watt, 288 mm long UV bulb  was used as the UV source. Standalone UV irradiation tests were conducted to determine the effect of UV irradiation on the test isolates with and without the second variable of freeze-thawing. As per previous studies, a 254 nm bulb was chosen as this wavelength is closest to the peak of DNA absorption and falls within the Martian UVC wavelength spectrum . Prior tE. coli strains were incubated at 37 \u00b0C for 18 h, while P. fungorum and D. radiodurans were incubated at 30 \u00b0C for 48 h and then enumerated. See The remaining 900 \u03bcL of the initial freeze-thaw test sample (detailed in GraphPad Prism (Version 8.3.0) software was used to analyze the data, which were represented in the form of colony forming unit (CFU) counts per aliquot measurement of sample recorded, whether for freeze-thaw cycle and/or UV treatment. Due to the large numbers of cells recorded, CFU/mL values were converted into a Log10 format. Initial starting CFU/mL concentrations were recorded for each biological repeat, for each test sample across each stage of experimentation and taking those values to be representative of 100% viability for each biological replicate. Standard deviation was depicted as +/\u2212 one standard deviation about the mean of the triplicate assays conducted for each test sample. A repeated measures analysis of variance (ANOVA) was conducted to investigate whether there was a bacterium, freeze-thaw cycle and/or UV energy effect on viability, whereby viability was the measure, freeze-thaw cycles and UVC energy were the within-subject factors and bacterium was the between subject factors. The assumptions of normality of the standardized residuals and sphericity were investigated and analysis was performed accordingly. These statistical analyses were all performed in SPSS Version 27.8 CFU/mL (E. coli ATCC 25922), 1.84 \u00d7 108 CFU/mL (E. coli DSM 9079), 3.28 \u00d7 108 CFU/mL (P. fungorum DSM 17061) and 3.92 \u00d7 107 CFU/mL (D. radiodurans ATCC 1393). The error bars depict +/- one standard deviation about the mean for each test sample. Notably, the percentage viability of E. coli 25922 was reduced by 71.3% after a single freeze-thaw cycle, while 10% viability was reached between freeze-thaw cycles 5 and 6. In contrast, the viability of E. coli DSM 9079 decreased by 50 % between freeze-thaw cycle 1 and 2, while 10% viability was observed between cycle 3 and 4. The percentage viability of P. fungorum DSM 17061 remained at 72.6% following a 20-freeze-thaw cycle period, while D. radiodurans ATCC 13939 exhibited a viability of 96% at the end of 20 successive rounds of freezing and thawing.In order to determine the viability of the selected test microbes when exposed solely to simulated Martian surface temperatures, successive freeze-thaw cycles were conducted in triplicate. E. coli strains with respect to themselves and the other test strains. Further analysis of E. coli ATCC 25922 showed a significant difference between freeze-thaw cycle 0 and freeze-thaw cycle 4 (p = 0.002), and again between cycle 8 and cycle 12 (p = 0.018), however, thereafter no significance was recorded in between cycles 8 to 12 (p = 0.304), 12 to 16 (p = 0.312) and 16 to 20 (p = 0.098). Statistical analysis of E. coli DSM 9079 showed significance between freeze-thaw cycles 0 to 4 (p = 0.0005), yet no significance from cycle 4 to 8 (p = 0.053), 8 to 12 (p = 0.204), 12 to 16 (p = 0.216) and 16 to 20 (p = 0.216). When subjected to statistical analysis, P. fungorum was found to be significantly different to each of the other test strains, yet no significance was recorded between freeze-thaw cycles 0 to 4 (p = 0.064) and 4 to 8 (p = 0.137), 8 to 12 (p = 0.250), 12 to 16 (p = 0.315) and 16 to 20 (p = 0.115). However, significance was seen when cycles 0 to 10 (p = 0.004) and 10 to 20 (p = 0.015) were statistically analyzed, thus showing a freeze-thaw effect over a longer range of time in comparison to the E. coli strains, which were more sensitive to the effects of freeze-thaw.Meanwhile, statistical analysis showed a difference between both D. radiodurans was found to be significantly different to each of the other test strains, yet no significance was recorded between freeze-thaw cycles 0 to 4 (p = 0.505), 4 to 8 (p = 0.809), 8 to 12 (p = 1.0), 12 to 16 (p = 0.862) and 16 to 20 (p = 0.718). Again, significance was not seen when cycles 0 to 10 (p \u2265 0.05) and 10 to 20 (p \u2265 0.05) were statistically analyzed, thus allowing for the conclusion that over the course of 20 freeze-thaw cycles, there is no long-term freeze-thaw effect seen on D. radiodurans.Statistical analysis, 7 CFU/mL (E. coli ATCC 25922), 1.88 \u00d7 108 CFU/mL (E. coli DSM 9079), 5.1 \u00d7 108 CFU/mL (P. fungorum DSM 17061) and 5.03 \u00d7 107 CFU/mL (D. radiodurans ATCC 1393). Each test sample was standardized to an OD600 nm reading of 1.0. The error bars depict +/\u2212 one standard deviation about the mean for each test sample. Experimentation was not conducted in monolayers in order to simulate how the microbial colonies would be found/behave on Mars if grown in biofilm-forming colonies on the surface or within bioreactor-like vessels. E. coli ATCC 25922 to independent UV treatment, with each point on the graph representing the average value of triplicate testing. Consequentially, the percentage viability of E. coli ATCC 25922 was reduced by 50% when exposed to 245 J/m2 of UVC irradiation. As expected, E. coli DSM 9079 exhibited even greater sensitivity to UVC irradiation with a 50% reduction in viability observed at a dose of 85 J/m2. When P. fungorum DSM 17061 was subjected to independent UV treatment the percentage viability of the sample was reduced by 50 % when exposed to 12,715 J/m2 of UVC irradiation. As depicted in D. radiodurans ATCC 13939 was exposed to independent UV treatment, the percentage viability of Deinococcus radiodurans ATCC 13939 was reduced by 50% upon exposure to 22,653 J/m2 of UVC irradiation.In order to determine the viability of the selected test microbes when exposed solely to simulated Martian UVC irradiation, independent UVC-dose-response-experimentation at 254 nm was performed. Treatments were conducted in triplicate, and the average CFU/mL counts of the three zero-time point test samples formed the basis for the average starting concentrations. When each strain was standardized to an OD600 nm of 1.0, the T0 standardized starting concentrations were 8.36 \u00d7 10Once survival parameters were determined for the individual treatments, the viability of the selected test microbes when exposed to a simulated combination of both Martian equatorial surface temperatures and UVC irradiation at 254 nm was determined. Successive freeze-thaw cycles and UVC dose response experiments were conducted in triplicate, using samples obtained from the individual freeze-thaw treatments. The average CFU/mL counts of the three zero-time point test samples formed the basis for the average starting concentrations.E. coli ATCC 25922 to successive rounds of freeze-thawing each followed by UV treatment. E. coli ATCC 25922 is a well-documented species of E. coli and has an intact SOS response system. This defined the parameters of survival when looking at the RecA- strain of E. coli. At freeze-thaw cycle 4, 8, 12, 16 and 20 , the percentage viability of E. coli ATCC 25922 was reduced by 50% at approximately 1270 J/m2, 1580 J/m2, 1060 J/m2, 1060 J/m2 and 1000 J/m2, respectively. E. coli DSM 9079 when exposed to successive rounds of freeze-thawing followed by UV treatment. At freeze-thaw cycle 4, 8, 12, 16 and 20, the percentage viability of E. coli DSM 9079 was reduced by 50% at approximately 2200 J/m2, 1060 J/m2, 430 J/m2, 215 J/m2 and 125 J/m2, respectively. D. radiodurans ATCC 13939 when subjected to successive rounds of freeze-thawing followed by UV treatment. As a true extremophile, it was expected to be the most resistant to Mars-like conditions of all the test microbes, thus setting the limits for UVC exposure. At freeze-thaw cycle 4, 8, 12, 16 and 20, the percentage viability of D. radiodurans ATCC 13939 was reduced by 50% at approximately 16,150 J/m2, 24,500 J/m2, 25,370 J/m2, 24,000 J/m2 and 23,500 J/m2, respectively. P. fungorum DSM 17061 to successive rounds of freeze-thawing followed by UV treatment. At freeze-thaw cycle 4, 8, 12, 16 and 20, the percentage viability of P. fungorum DSM 17061 was reduced by 50% at approximately 3800 J/m2, 5850 J/m2, 5000 J/m2, 4500 J/m2 and 4300 J/m2, respectively.The effects of Mars-like environmental conditions (freeze-thaw and UVC radiation) on bioremedial microbial strains was investigated to determine the limits that microbial life can withstand in a Mars-like environment. It is known that freezing and thawing impose several interconnected stresses on cells, including dehydration, hyperosmotic stress, ice formation and oxidative stress. In addition to this, other variables such as nutrient availability and the rate of cooling all play a role in cellular viability within a freeze-thaw environment ,25,26. TE. coli ATCC 25922 can withstand at least 20 successive freeze thaw cycles. While an average reduction in viability of 71.3% was observed between cycle 0 and cycle 1, the remaining 28.7% of the population were able to withstand successive cycles in the absence of a cryoprotectant with a gradual decline in viability being observed over time. While it is known that the formation of intracellular ice crystals can cause cells to lyse completely, there is also a molecular mechanism to this process [E. coli (E. coli B/r), in the absence of a cryoprotectant, was used in the 1975 study and only a single freeze-thaw cycle (unlike this study where 20 freeze-thaw cycles were examined) was observed with temperatures ranging from \u2212196 \u00b0C to +25 \u00b0C. Similarly, a study conducted by Lyscov based on the intrinsic viscosity and sedimentation constants of calf-thymus DNA and phage-T2 DNA, showed that a decrease in molecular weights of the DNA samples was caused by freeze-thaw, while another study by Shao determined that progressive DNA degradation of blood samples was observed in samples with DNA sizes larger than 100 kb, as these were found to be most sensitive to freeze/thaw degradation. It was also seen that increasing the DNA concentration of the stored samples from 10 \u03bcg/mL to 100 \u03bcg/mL had a protective effect on DNA stability [ process . As demotability ,30.E. coli DSM 9079, as seen in E. coli DSM 9079 acted as the SOS-repair-system-deficient control within this study. E. coli DSM 9079 was a \u0394recA mutant, while E. coli ATCC 25922 had the functional RecA gene. RecA encodes a multifunctional enzyme that plays an important role in DNA repair, homologous recombination and the induction of the SOS biomolecular response systems within the cell [E. coli ATCC 25922, over the course of the successive freeze-thaw cycles, it is evident that absence of the RecA gene played an important role in the survivability of E. coli DSM 9079 in response to freeze thawing. When compared to the more robust control (E. coli ATCC 25922), which survived 20 successive freeze-thaw cycles, the \u0394recA mutant was non-viable by freeze-thaw cycle 9. Statistical analysis determined that there were differences amongst both E. coli strains when subjected to repeated freeze-cycles, as stated in the results. While genomic studies are indicated, it could be hypothesized that DNA repair systems are activated after this primary freeze-thaw cycle, owing to the prolonged survival of the remaining 28.7% of E. coli ATCC 25922 cells .When examining the percentage viability of the cell . While nE. coli B cell-line progeny cells, which had been exposed to daily freeze-thaw cycles, developed on average a 54% mortality rate to freeze-thaw conditions compared to their less sensitive ancestors that showed an average mortality rate of 34% [E. coli DSM 9079, unexpectedly, exhibited an average initial decrease in viability of 28%, whereas E. coli ATCC 25922 had an average initial decrease of 71.3%, although was recoverable in small numbers after 20 cycles.Conversely, it has been shown in another study that e of 34% . It is nP. fungorum DSM 17061 was exposed to the same set of freeze-thaw conditions as both the robust and sensitive controls, it exhibited an increased resistance to the effects of freeze-thaw. Indeed, a negligible average initial decrease in viability between cycle 0 and cycle 1 of 2% was observed, and over the course of 20 cycles, the 50% viability threshold had not been reached. This strain exhibited an average viability of 76%; meaning it is capable of withstanding successive freeze-thaw cycles without a dramatic loss in cell population when beginning with a starting concentration of 3.28 \u00d7 108 CFU/mL. Based on these data, it could be suggested that due to the relatively large genome size of P. fungorum DSM 17061 , the studies . In addi by Shao .D. radiodurans ATCC 13939 is a well-documented bacterium in the field of astrobiology, largely due to its renowned radio-resistance. It has been shown that D. radiodurans uses redundant \u2018multi-genomic\u2019 copies to aid in DNA repair by means of inter-chromosomal recombination [P. fungorum DSM 17061, the 50% viability threshold was not reached over the course of the 20 cycles. Remarkably, after the last experimental cycle (cycle 20) the average cell viability was 96%. When the data are compared to the findings of Alur and Grecz (1975), where E. coli B/r was subjected to a single freeze-thaw cycle with a temperature range of \u2212196 \u00b0C to 25 \u00b0C (which concluded that freeze-thaw causes DNA degradation), the data recorded here further supports their research due to the known efficiency of D. radiodurans DNA repair systems, and its evidential ability to survive successive freeze-thaw cycles. If multiple genomes and highly efficient DNA repair systems are key in surviving the effects of freeze-thaw, it could be suggested that P. fungorum DSM 17061\u2032s single yet large genome, may encode repair systems that are more efficient than the robust control E. coli strain, although less efficient than D. radiodurans multiple genomes. Statistical analysis helps to further support the hypothesis that P. fungorum is just as resistant to external environmental insults as D. radiodurans, as will be discussed further on in this paper.bination ,35, and E. coli ATCC strain 25922, E. coli DSM strain 9079, P. fungorum DSM strain 17061 and D. radiodurans ATCC strain 13939. Recorded bacterial strains exposed to freeze-thaw experimentation within the published literature include E. coli B [E. coli can tolerate freeze-thaw to varying degrees, however it must be stated, that in this study, cells were slowly thawed over 8 h and heated to 30 \u00b0C, whereas Sleight et al., thawed at room temperature (22 \u00b0C) for 1.5 h, thus not allowing for direct comparisons. Interestingly, when Pseudomonas paucimobilis [E. coli strains, with sustained levels thereafter, and also evident with D. radiodurans and P. fungorum.While examining the effects of freeze-thaw, it is important to highlight the importance of the work carried out in furthering our understanding of how these microbes react to extra-terrestrial conditions, as there is no available literature focused solely on the long-term independent freeze-thaw cycling of . coli B . It was imobilis  was expoE. coli strains, it is possible that the lack of a DNA repair system within E. coli DSM 9079 may upregulate another response pathway attributing to an initial reduction in freeze-thaw mediated cellular damage during the first freeze-thaw cycle, such as the induction of a nucleotide excision repair system consisting of the genes uvrA, uvrB, uvrC and uvrD [E. coli ATCC 25922, which experienced a dramatic initial decrease based on the action of a competent cell regulatory system in response to damage, yet sustained numbers throughout unlike E. coli DSM 9079. One study notes that low temperature and long storage time could induce the incomplete disassembly of formed RecA-DNA nucleoprotein filaments, thus causing a failure in cell repair in cells with fully functioning repair systems [While examining the effects of freeze-thaw on both and uvrD ; in comp systems , which mP. fungorum has shown itself to be equally resistant to environmental insults as D. radiodurans, this may be due to DNA compaction. It is important to note, the nucleoid of radioresistant microbes, such as D. radiodurans, are known to be highly condensed structures, which allow the organism to withstand high doses of irradiation (DNA damage) while remaining unaltered [D. radiodurans. P. fungorum was also determined to be a highly UV resistant microbe, and while there is no published literature to support the existence of HU protein in P. fungorum, it has been shown that Paraburkholderia cenocepacia (a closely related strain) expresses HU protein [P. fungorum and D. radiodurans may facilitate a tolerance to DNA degradation caused by freeze-thaw conditions.While hypothesizing why naltered . This is protein . TherefoE. coli ATCC 25922, there are multiple studies demonstrating the production of heat-shock proteins in response to temperatures of 45 \u00b0C and above [E. coli genus is known to be a carrier of cold-shock proteins [E. coli ATCC 25922, there is a strong possibility that cold-shock proteins are also playing a role in the bacterium\u2019s prolonged survival. In both proof of concept, and in demonstration of the converse, RecA has been implicated as a cold-shock inducing protein [E. coli DSM 9079 (a \u0394recA mutant) exhibits reduced viability in response to freeze-thaw cycling. Again, while not directly comparable due to the speciation differences, it is possible that cold-shock proteins, or their absence, could play a role in the extent of the observed viability. Previous studies have shown that D. radiodurans is capable of producing both heat-shock and cold-shock proteins. One study found an increased expression of 67 proteins during heat-shock and 42 proteins during cold-shock  and autoradiography) [P. fungorum genome has also revealed the presence of cold-shock proteins [Microbial molecular adaptations to the cold include slow chemical reaction rates and limited enzyme activity, denaturation of proteins, increased water viscosity, decreased cell membrane fluidity and limited water availability for biochemical reactions ,43,44,45nd above ,49. Whilproteins , therefo protein . E. coliography) . It mustproteins . When coE. coli, Paraburkholderia, and Deinococcus [Pseudomonas, Arthrobacter, Flavobacterium and Deinococcus) were able to change their lipid membrane structures in order to retain fluidity depending on the temperature. It was found that this fatty acid distribution was also affected significantly by the growth temperature of the surrounding environment. It is noted that there is no testing completed to date on the effects of membrane lipid distribution during freeze-thaw and UVC irradiation.While not greatly studied in a diverse range of prokaryotic organisms, the process of homeoviscous adaptation may also play a role in the prolonged survival of these microbes. Microbes that are capable of synthesizing their own fatty acids include nococcus ,55,56. Tnococcus ,57,58,59The effect of independent exposure to damaging UVC irradiation was investigated to observe how these bacterial strains behave when exposed to UVC irradiation similar to the Martian environment (200\u2013280 nm). The results were intended to inform future studies into the suitability and longevity of bacteria within extra-terrestrial habitats when exposed to UVC irradiation at 254 nm.E. coli ATCC 25922 was exposed to 245 J/m2, resulting in a standard dose response curve. When compared to E. coli DSM 9079, over four times the amount of UVC energy was needed to inactivate E. coli ATCC 25922 compared to the \u0394recA mutant E. coli DSM 9079, which required a UVC dose of 85 J/m2 in order produce a 50% reduction in viability. Again, while not genetically identical, this finding reaffirms the important function of the RecA gene in DNA repair/homologous recombination and the SOS biomolecular response systems. These systems are needed as UVC energy is absorbed by DNA causing the dimerization of nucleic acid bases including cyclobutane pyrimidine species and pyrimidine (6\u20134) pyrimidone compounds, together with their Dewar isomers [RecA+ strain (E. coli ATCC 25922), they denote a \u201csensitive\u201d and \u201ccompetent SOS response\u201d range for judging the viability of the test strains when exposed to simulated Mars-like conditions. They allow for the creation of a guideline of the lower and upper limits of survival for the average terrestrial microbe.rimidine \u20134 pyrimiP. fungorum DSM 17061. It was noted that, experimentally, 10,000 J/m2 of UVC energy was required to reduce the viability of P. fungorum DSM 17061 by 50 %, with 44,415 J/m2 of UVC energy being required to inactivate the sample completely. When the UVC resistance of P. fungorum DSM 17061 was compared to those of a known radio-resistant bacterial strain (D. radiodurans ATCC 13939), 25,000 J/m2 of UVC energy was required to reduce viability by 50 %, with complete inactivation of the test sample observed at 50,760 J/m2. When the death points of both species were compared, a difference of 6345 J/m2 was recorded, suggesting that P. fungorum DSM 17061 is a highly UV-resistant micro-organism, which has not been reported before.P. fungorum and D. radiodurans in response to comparable UVC energy doses (p = 0.685). The lack of significance denotes that while both species are not totally comparable, they are not statistically dissimilar to each other. This further supports the hypothesis that P. fungorum is akin to D. radiodurans in its ability to withstand extreme environmental insults. It is noted that a 254 nm wavelength was chosen for both its maximal absorbance by DNA and based upon its lack of photoreactive properties. Photoreactivation is the phenomenon by which UV-inactivated organisms regain their activity via photorepair of UV-induced lesions in the DNA by utilizing the energy of near-UV light (310\u2013480 nm) and an enzyme, photolyase [When subjected to statistical analysis, a significant difference was not seen in the recorded viabilities of otolyase ,61,62.E. coli at 254 nm based upon photoreactivation found that wavelengths from 220 nm to 300 nm reduced the level of photorepair in pyrimidine dimers due to a possible interference of the photolyase repair enzyme. As a result, the tests carried out in this paper were subjected to low levels of photorepair occurrences.While photolyases allow for the mitigation of UV damage, most bacteria have either a single photolyase or photolyase-like gene with varying expression levels . StudiesD. radiodurans is one of the most radiation resistant life forms on the planet and as a result is often used as a positive control to demonstrate a high level of radiation resistance within an experimental study [UV resistance is has also been linked to a high intracellular ratio of manganese (Mn) and iron (Fe). Interestingly, this phenomenon is proven to be irrespective of the surrounding levels of Mn and Fe within the environment . D. radial study . Interesal study .P. fungorum, in addition to the possibility of DNA compaction as discussed earlier. Despite the paucity of research on this species, the identification of a type VI secretion system, T6SS-4, is implicated in the metal acquisition of intracellular manganese to protect from oxidative stress within the species Paraburkholderia thailandensis [Based on these data, it is possible that an abundance of intracellular Mn and Fe may be contributing to the UVC resistance of andensis .E. coli as a species does not routinely import Mn into the cell, although it has been shown to do so when iron is unavailable. This allows Mn to act as a substitute for iron, a common enzyme co-factor [RecA plays no part within the induction of Mn-superoxide-dismutase (which prevents the oxidative stress caused by high Mn levels) [o-factor . It doeso-factor . Interes levels) .E. coli strains and different bands of the UV spectrum consistently illustrate the effects of growth phase on UV resistance. It has been demonstrated how E. coli strains show a significant resistance to UV-induced damage when in stationary phase, as opposed to when exponentially growing during log phase [When the effects of the combined treatment of both freeze-thaw and UVC irradiation on the chosen bacterial strains was investigated, a species-dependent synergistic and antagonistic effect was observed. As mentioned, UVC damages active cells by inducing changes in the chemical structure of the DNA strands, resulting in the production of cyclobutane pyrimidine dimers. These dimers cause DNA molecule distortion, which in turns causes defects in DNA replication, which can lead to cell death ,71. In aog phase ,74. As s2 of energy was required to reduce the viability of E. coli ATCC 25922 by 50%  where a significant reduction in UV inactivation was observed in cells that had been frozen prior to UVC irradiation (UV fluences of 15 to 90 J/m2 were used) with E. coli ATCC 25922 showing more UV resistance than the other test organism E. coli O157:H strain 961019 [Upon independent exposure to UVC energy, 200 J/m2 by 50% . When th2 by 50% A, at freE. coli DSM 9079 was independently exposed to UVC energy, a dose of 75 J/m2 was required to reduce viability by 50%. Similarly, when exposed to a combination of both successive freeze-thaws and UV irradiation  and E. coli all have reported doubling times of 2.6 h, 3 h and 25 min, respectively [E. coli strains showed increased levels of viability, while both resistant strains (D. radiodurans and P. fungorum) showed a reduced viability with respect to the independent insults.When examining the combined effects of freeze-thaw and UV, it is important to note the doubling times of the test candidates. ectively ,77,78. WWhile UV resistance upon freezing and thawing appears to be strain-dependent, this phenomenon is useful for microbes intended for bioremediation in extra-terrestrial environments. There are no reported studies examining the phenotypic response of the selected strains to UVC energy over the course of 20 freeze-thaw cycles, thus highlighting a level of importance in furthering our understanding of these microbes both terrestrially and extra-terrestrially.E. coli ATCC strain 25922, E. coli ATCC strain 9079, P. fungorum DSM strain 17061 and D. radiodurans ATCC strain 13939 were subjected to three different rounds of experimentation to simulate Mars-like conditions. These included individual successive freezing and thawing experiments, with temperatures ranging from \u221280 \u00b0C to +30 \u00b0C, independent UVC dose-response experimentation and a combination of both freeze-thaw followed by UVC exposure. When exposed to these parameters separately, the test samples responded as detailed above, however, when combined, both species-dependent synergistic and antagonistic effects were observed.In this study E. coli strains, while reducing the UVC-resistance of P. fungorum and D. radiodurans. It was also observed that P. fungorum DSM 17061 is a highly UV-resistant microbe. Given the resistant nature of the test microbes to adverse Mars-like conditions, these species are potential candidate bioremedial microbes for future potential employment on Mars, based upon their Earthly bioremedial capabilities.While the results observed are species-dependent, the overall experimental data suggest that freeze-thawing significantly increases the UV resistance of both Within an extra-terrestrial setting, these data suggest that a wide variety of common bacteria can survive the tested immediate harsh conditions found on the equatorial Mars-like landscape. Using the data obtained from this research, these selected microbes could potentially be capable of being strategically introduced to the future Martian landscape."}
+{"text": "Medical and Health Organization (MHO) staff\u2019s emergency preparedness awareness and behaviors are essential variables that affect public health emergency response effectiveness. Based on the theory of psychological capital and the theory of planned behavior (TPB), this study discusses the mechanism of the psychological characteristics of MHO staff on their emergency preparedness behavioral intention (EPBI). To verify the research model, we conducted a web-based questionnaire survey among 243 MHO staff from China and analyzed the data using the structural equation modeling software, AMOS 24.0 . The empirical results reveal that psychological capital significantly affected cognitive processes theorized by TPB. This study suggests that the positive psychological capital of MHO staff should be developed and managed to improve their EPBI. With deterioration of the natural environment, changes in the international situation, and the acceleration of domestic, economic, and social reform, the frequency of various natural disasters, accident disasters, mass incidents, and public health events has shown a significant upward trend ,2,3,4. TEmergency preparedness is a sub-field of emergency management, mainly referring to the establishment and maintenance of various preparations for emergency prevention, early warning, response, and recovery. It includes emergency plan formulation, personnel training, preparation and custody of emergency supplies and equipment, drills within the emergency plan, and connection with external emergency forces. Its ultimate goal is to maintain the emergency capacity and rapid response capacity needed for emergency rescue in relation to significant accidents and ultimately reduce casualties and unnecessary losses . The MedIn order to fill the gap in this field, we try to use the theory of planned behavior (TPB) to understand the self-driving mechanism of MHO staff\u2019s emergency preparedness behavioral intention (EPBI). In this study, EPBI is considered to be the intention of MHO staff to prepare so as to avoid losses from emergencies. As an essential theory to explain the general decision-making process of individual rational behavior, TPB is widely used in the field of behavioral science, and has been proved to have good explanatory and predictive power of human behavior, and can help researchers understand how people change their behavior patterns ,12,13. TTherefore, we attempt to add psychological capital (PsyCap) as an antecedent factor to the TPB model in order to better understand the formation mechanism of MHO staff\u2019s EPBI, and simultaneously stimulate their subjective initiative to participate in emergency preparedness work. The psychological capital proposed by the American management scientist Luthans et al. is regarded as a positive psychological state in the process of individual growth and development, a core psychological element beyond human capital and social capital, and a psychological resource to promote personal growth and performance improvement, including hope, optimism, resilience, and self-efficacy ,18,19. AIn this study, attitude refers to MHO staff\u2019s evaluation of their psychological tendency to conduct emergency preparedness. Perceived behavioral control refers to the difficulty or ease that MHO staff feel when responding to emergency preparedness. Finally, subjective norms refer to the social pressure that MHO staff feel when deciding whether to conduct emergency preparedness, primarily obtained by consulting or observing others\u2019 behavior .Firstly, TPB believes that an individual\u2019s attitude towards behavior will affect his or her behavioral intention. In a specific time and environment, individuals can acquire a small amount of beliefs about behavior, which are the cognitive and emotional basis of attitude. Among them, individuals with positive beliefs and values about emergency-related content, knowledge, and skills tend to participate in emergency prevention and preparedness, such as emergency knowledge popularization activities and emergency training drills . Those wHypothesis\u00a01\u00a0(H1).MHO staff\u2019s attitude towards emergency preparedness behavior has a positive impact on their EPBI.Secondly, TPB also believes that perceived behavioral control is related to behavior intention. Perceived behavioral control also emphasizes an individual\u2019s ability to cope with tasks or make choices to a certain extent, and this ability perception mainly comes from a sense of self-efficacy . PreviouHypothesis\u00a02\u00a0(H2).MHO staff\u2019s perceived behavioral control has a positive impact on their EPBI.Finally, an individual\u2019s behavior is influenced or motivated by the norms observed in their environment. For example, before the disaster, if the MHO staff noticed that the people around them  were making preventive preparations, they were more likely to participate actively in emergency prevention and preparedness. Conversely, subjective norms also reflect the degree of support of external factors for MHO staff\u2019s emergency preparedness behavior to a certain extent and play a vital role in the formation of individual emergency attitudes and perceived behavioral control ,43. For Hypothesis\u00a03\u00a0(H3).The subjective norms of MHOstaff will have a positive impact on their EPBI.Hypothesis\u00a04\u00a0(H4).The subjective norms of MHOstaff will have a positive impact on their attitude towards emergency preparedness behavior.Hypothesis\u00a05\u00a0(H5).The subjective norms of MHOstaff will have a positive impact on their perceived behavioral control.Positive PsyCap reflects the following view: Firstly, regardless of whether they are facing the disaster threat, MHO staff are willing to carry out all kinds of emergency preparedness work . SecondlHypothesis\u00a06\u00a0(H6).MHOstaff\u2019s PsyCap has a positive impact on their attitude towardsemergency preparednessbehavior.In recent years, through factor analysis, researchers have found that the standard of perceived behavioral control is loaded on two factors. The former reflects the belief in self-efficacy , and the latter reflects the belief in control  . HoweverHypothesis\u00a07\u00a0(H7).MHOstaff\u2019s PsyCaphas a positive impact on their perceived behavioral control.As an effective way to enhance inner strength and promote individual growth, PsyCap can help MHO staff adjust to psychological and physical problems caused by inter-personal relationships and work stress and improve personal trust and satisfaction ,48. TherHypothesis\u00a08\u00a0(H8).MHOstaff\u2019sPsyCaphas a positive impact on their subjective norms.With careful consideration of Hypothesis 1 to Hypothesis 8, this study puts forward the intermediary hypothesis of attitude, perceived behavioral control, and subjective norms. Based on the previous discussion, this study holds that the positive PsyCap of MHO staff will have a positive impact on their attitude towards emergency preparedness behavior, perceived behavioral control and subjective norms. And these factors will also positively affect their EPBI. Therefore, we have reason to expect that MHO staff\u2019s attitude, perceived behavioral control and subjective norms may play an intermediary role between PsyCap and EPBI. In addition, we also believe that MHO employees\u2019 subjective norms will promote their attitude and perceived behavioral control, and this study also proposes the intermediary role of attitude and perceived behavioral control between subjective norms and EPBI. The specific assumptions are as follows:Hypothesis\u00a09\u00a0(H9).The attitude towardsemergency preparedness behaviorof MHOstaff acts as an intermediary between PsyCap and EPBI.Hypothesis\u00a010\u00a0(H10).The perceived behavioral controlof MHO staff acts as an intermediary between PsyCap and EPBI.Hypothesis\u00a011\u00a0(H11).The subjective norms of MHOstaff act as intermediaries between PsyCap and EPBI.Hypothesis\u00a012\u00a0(H12).The attitude towards emergency preparedness behavior of MHO staff acts as an intermediary between subjective norms and EPBI.Hypothesis\u00a013\u00a0(H13).The perceived behavioral control of MHO staff acts as an intermediary between subjective norms and EPBI.We used the method of questionnaire survey to test the research model. The data sources of this study were accurate and reliable. Firstly, to avoid the systematic error caused by the deviation of the standard method, this study invited five doctoral students of related majors to revise the questionnaire repeatedly to make the question items as concise and easy to understand as possible. To avoid individual repetition, we set the questionnaire to be answered only once per IP address. Secondly, to encourage the participants to answer the questions frankly and truthfully, the online questionnaire used in this study provided complete anonymity\u2014the researchers never knew the identity of the interviewees. Further, the survey was conducted and analyzed outside the organization\u2014enhancing the interviewees\u2019 perceived anonymity and actual anonymity. Finally, to ensure the diversity of data sources, this study selected a group of staff composed of staff from different MHOs in China as the research cohort. The use of group samples increases the certainty that the sampling population will accurately represent the target population, and the survey subjects are MHO staff. Therefore, the survey results are more likely to be extended to all MHO workers\u2019 groups.The measurement scales used in this study were adapted from the maturity scale proposed by previous scholars. We invited relevant professionals to translate repeatedly to avoid measurement errors caused by semantic differences. Considering that if a potential variable is measured by three or more observation variables, the estimation deviation of the model parameters is almost zero, this study retained three questions for each potential variable. The measurement items of PsyCap were adapted from the questionnaire of Luthans et al. (2007) . The resInitially, the survey received responses from 289 MHO staff. After excluding incomplete answers and screening out spoiled solutions , it was determined that the number of valid samples was 243, and the effective recovery rate was 84.1%. Among the participants, 80.7% were female, 37.4% were aged between 18 to 24, 72.1% had a Bachelor\u2019s degree or above, and 45.7% were nurses. Additionally, 40.7% of the respondents supported Wuhan during the \u201cCOVID-19\u201d epidemic. In this study, we analyzed the data in three steps. In the first stage, the reliability and validity of the measurement model were tested. In the second stage, the fitness of the structural equation model was tested. In the third stage, the structural equation model was used to test the hypotheses. SPSS 26.0  and AMOS 24.0  were used to analyze data.Although all the scales in this study have been recommended, their reliability and validity still need to be evaluated. Firstly, SPSS 26.0 was used to test the reliability of the questionnaire data. The results show that the Cronbach\u2019s \u03b1 coefficient of the whole questionnaire data is 0.960, and that of each variable is more significant than 0.8, indicating that the internal consistency of the scale used in this study is promising.Secondly, we use AMOS 24.0 software to analyze the validity of the questionnaire data, including the content validity test, convergent validity test, and discriminant validity test. The results show the following: First, except for one observation variable\u2019s standard factor load coefficient between 0.6 and 0.7, the other observation variables are all above 0.7; all of them have reached a significant level, indicating that the questionnaire has good content validity. Second, the composite reliability (CR) of each variable is more significant than 0.8, and the average variance extracted (AVE) is more significant than 0.6, indicating that the scale has good convergent validity. Third, there is no obvious distinction between the four substructures of PsyCap, but there is obvious differentiation between these four substructures and other variables, as well as other variables. Previous studies have conceptualized PsyCap into a higher-order structure . Compare2/df, RMR, RMSEA, and other indicators to verify the fit of the model and data. The results showed that all the indexes reached or approached the range of recommended standards , and there was a good fit between the model and the data. After testing, we found the absolute value of skewness of each observed variable was between 0.155 and 1.140, and the absolute value of kurtosis was between 0.022 and 3.520, so the data formed a normal distribution. The Chi-square versus Mahalanobis distance diagrams of variables were drawn by using the extension program \u201cNormaltest_V1.0\u201d of SPSS 26.0 software. The points in the map approximately formed a straight line, and the combination of all observed variables was close to multivariate normal distribution, so the maximum likelihood estimation method was used to estimate the model parameters. Considering that the overall fitting index value of the model is easily affected by the number of samples, this study selected \u03c7p < 0.001) and perceived behavioral control  significantly affected their EPBI, supporting Hypothesis 1 and Hypothesis 2. Furthermore, the subjective norms of MHO staff significantly affected their attitude  and perceived behavioral control , Hypotheses 4 and 5 were supported. Finally, the PsyCap of MHO staff had a significant influence on their attitude towards emergency preparedness behavior , perceived behavioral control , and subjective norm , Hypotheses 6\u20138 were supported. However, the subjective norms of MHO staff  did not significantly impact their EPBI, and Hypothesis 3 was not supported. A reasonable reason for this result may be bias in the study design. For example, there is no unified consensus on the definition of subjective norms in this study. However, this study mainly adopts mandatory subjective norms, whether the essential people around them support their EPBI, but does not consider the demonstrative subjective normative structure, such as how the important people around them do it themselves. Conversely, it may be because most human behaviors are under the control of self-will; thus, the EPBI of MHO staff is more affected by their attitude and perceived sense of behavioral control, making the role of subjective norms relatively weak.Based on the premise that the above model fits well, we tested the hypothesis of this study, and p-value is 0.040. The results show that the indirect effect is significant, and its estimated value is 0.122, supporting Hypothesis 9. The confidence interval of intermediary path \u2461 under the Bias-Corrected method at 95% confidence level is , and the p-value is 0.003. The results show that the indirect effect is significant, and its estimated value is 0.113, supporting Hypothesis 10. According to the same method of judgment, we found that paths \u2463 and \u2464 were also significant, supporting Hypotheses 12 and 13. However, the confidence interval of intermediary path \u2462 under the Bias-Corrected method at 95% confidence level is , and the p-value is 0.303. The results show that the indirect effect is not significant, Hypothesis 11 was not supported, echoing the fact that Hypothesis 3 is not valid in the hypothesis test.In testing the mediating effect, we consider that the commonly used Sobel method has some limitations. Therefore, this study chooses the bootstrap method to test the mediating role of attitude, perceived behavioral control, and subjective norms in the three paths based on 5000 repeated samplings ,58. TablWith the continuous spread of global social risks, the emergency management system plays an increasingly important role in China\u2019s national governance system . HoweverFirstly, in the context of China, TPB can be used to predict and explain the intention of emergency preparedness behavior, which further expands the application field of TPB. Secondly, external motivation and internal opportunity work together to affect the EPBI of MHO staff. Most of the previous literature considered the influence of external environmental factors on the formation of EPBI, but seldom considered the influence of individual internal factors. However, the impact of external drivers on EPBI is not real-time and effective, which largely depends on individual self-management. Therefore, this study integrates the theoretical viewpoints in the field of positive psychology and organizational behavior, and further discusses the self-driving mechanism of MHO staff\u2019s EPBI. Talking about the influence of PsyCap on EPBI enriches the existing research literature in the field of emergency preparedness, and its conclusions deepen our understanding of the formation mechanism of EPBI. Finally, this study complements the original TPB model by including role-width resources that can promote individuals to perform a broader range of tasks, which provides a more perfect theoretical model for predicting role-width behavior in MHO staff\u2019s behavior database and provides an important update consideration for the development of a comprehensive behavioral model.(1)Cultivate crisis awareness and improve the psychological risk reserve of MHO staff.(2)Strengthen the training of emergency knowledge to make MHO staff fully aware of the significance and value of EP.(3)Conduct emergency practice drills to enhance the confidence of MHO staff in dealing with unexpected accidents.(4)Establish emergency logistics support work to ensure MHO workers\u2019 health and life safety, etc.Understanding the influence of TPB on EPBI and the influence of PsyCap on behavioral decision-making process are of great significance to the practical application of positive psychology and organizational behavior theory. Firstly, considering the impact of TPB mechanism on EPBI, management can take corresponding measures to encourage MHO staff to develop the necessary safety skills and knowledge, and stimulate their EPBI in multiple ways. These measures are as follows:(1)Involve MHO staff in the process of preparing emergency preparedness and response plans.(2)Make realistic and optimistic expectations to counteract the pessimism of MHO staff about emergency preparedness.(3)Reinforce the transferable value of emergency preparation behavior in the career development of MHO staff.(4)Provide positive feedback to MHO staff who are actively involved in emergency preparedness, etc.Additionally, the influence of PsyCap on the behavioral decision-making process also provides a unique opportunity for managers to improve the enthusiasm of MHO staff to carry out emergency preparedness work. Managers can develop the PsyCap of MHO staff at the sub-structure or macro-level to improve their attitude towards emergency preparedness behavior, perceived behavioral control and subjective norms. The following are several measures to develop the PsyCap of MHO staff:This study has important theoretical and practical significance for understanding and stimulating MHO staff\u2019s EPBI, but there are still some limitations. First, the subjects of this study are MHO staff, and the original data were collected through the form of a network questionnaire; hence, the homologous bias cannot be eliminated. Therefore, the research method should be further improved in subsequent studies. Second, the model in this study only considered the mediating effect and did not involve moderating variables. Therefore, future studies should consider more influencing variables to make the results more objective and comprehensive. Finally, the samples used in this study to verify the hypothesis are from China, and whether the research results can be inferred in other countries (regions) needs to be further verified. Follow-up research can increase the scope of the survey and sample size, so that the research results can adapt to a wider range of research objects, and improve the external validity of the research results."}
+{"text": "In vitro activities of ceftazidime-avibactam (CAZ-AVI) and key comparators against AmpC-overproducing Enterobacterales and Pseudomonas aeruginosa isolates from four Phase 3 clinical trials and against OXA-48\u2013producing Enterobacterales with multiple resistance mechanisms from the Antimicrobial Testing Leadership and Surveillance (ATLAS) program were evaluated. Susceptibility to CAZ-AVI and comparators was determined by reference broth microdilution methods. Clinical response at test of cure (TOC) was assessed in patients from Phase 3 trials with baseline OXA-48\u2013producing Enterobacterales or AmpC-overproducing Enterobacterales and P. aeruginosa treated with CAZ-AVI or comparators. Against 77 AmpC-overproducing Enterobacterales isolates from Phase 3 trials, meropenem-vaborbactam (98.7% susceptible [S]), CAZ-AVI (96.1% S), and meropenem (96.1% S) had similar in vitro activity and were more active than ceftolozane-tazobactam (24.7% S). Clinical cure rates in patients with baseline AmpC-overproducing Enterobacterales were 80.7% (n\u2009=\u200921/26) and 85.0% (n\u2009=\u200917/20) for CAZ-AVI and comparators. Against 53 AmpC-overproducing P. aeruginosa isolates from Phase 3 trials, CAZ-AVI (73.6% S) was more active in vitro than ceftolozane-tazobactam (58.5% S) and meropenem (37.7% S). Clinical cure rates in patients with baseline AmpC-overproducing P. aeruginosa were 85.7% (n\u2009=\u200912/14) and 75.0% (n\u2009=\u20099/12) for CAZ-AVI and comparators, respectively. Of 113 OXA-48\u2013producing isolates from the ATLAS program, 99.1% were susceptible to CAZ-AVI. Four patients with baseline OXA-48\u2013producing Klebsiella pneumoniae isolates treated with CAZ-AVI in Phase 3 trials were clinical cures at TOC and had favorable microbiological response. CAZ-AVI was among the most active agents against AmpC-overproducing P. aeruginosa and Enterobacterales and had greater in vitro activity against OXA-48\u2013producing Enterobacterales than meropenem-vaborbactam, meropenem, ceftolozane-tazobactam, and other comparators. Enterobacterales and Pseudomonas aeruginosa are the most common Gram-negative organisms causing serious bacterial infections with high morbidity and mortality. When these organisms produce either AmpC or OXA-48 \u03b2-lactamases, in combination with extended-spectrum \u03b2-lactamases (ESBLs), they are often resistant to most \u03b2-lactam antibiotics, resulting in very few treatment options and higher mortality , as well as the constitutively expressed plasmid-encoded blaampC genes (pAmpC)  . A stable system . Strainsfailures , 6. PathCeftazidime-avibactam (CAZ-AVI) is a combination of the \u03b2-lactamase inhibitor avibactam and the broad-spectrum cephalosporin ceftazidime. Avibactam is a non\u2013\u03b2-lactam \u03b2-lactamase inhibitor that binds reversibly to \u03b2-lactamases, efficiently inactivating \u03b2-lactamase enzymes and preventing the hydrolysis of \u03b2-lactam compounds, such as ceftazidime . CAZ-AVIin vitro activities of CAZ-AVI and key comparators against AmpC-overproducing Enterobacterales and P. aeruginosa isolates collected from Phase 3 clinical trials, RECLAIM , complicated urinary tract infections (cUTI), or hospital-acquired pneumonia/ventilator-associated pneumonia (HAP/VAP) caused by Gram-negative pathogens. REPRISE was an open-label clinical study that evaluated the efficacy of CAZ-AVI compared with best available therapy (primarily carbapenems) in patients with either cIAI or cUTI caused by ceftazidime-resistant Gram-negative pathogens. OXA-48\u2013producing Enterobacterales with multiple resistance mechanisms were also collected from a CAZ-AVI global surveillance program , and susceptibility of these isolates to CAZ-AVI and comparator antibiotics was tested as part of this study. Finally, clinical response at test of cure (TOC) was also assessed for patients from the Phase 3 studies that had AmpC-overproducing or OXA-48\u2013producing Enterobacterales and P. aeruginosa at baseline.This study evaluated  RECLAIM , RECAPTU RECLAIM , REPROVE RECLAIM , and REP RECLAIM  , followed by amikacin , meropenem-vaborbactam (MVB) , and gentamicin . Overall, the in vitro activities were poor for ceftazidime , meropenem , levofloxacin , and ceftolozane-tazobactam  against OXA-48\u2013producing Enterobacterales . It is important to note that CAZ-AVI (98.4% S) remained highly active against OXA-48\u2009+\u2009ESBL + limited-spectrum \u03b2-lactamase (LBSL)-coharboring isolates. These isolates had decreased susceptibility to MVB (68.9% S), ceftazidime (18.0% S), MEM (8.2% S), and TZC (4.9% S) with no major changes in susceptibility to AMK  and was very limited in MEM and TZC . The most common ESBL gene types were the K 82% S; . PlasmidK. pneumoniae possessing OXA-48bla in combination with additional \u03b2-lactamases . At baseline, the CAZ-AVI MIC for all isolates was 0.5\u20131\u2009mg/L and the ceftazidime MIC was >64\u2009mg/L. All 4 K. pneumoniae isolates were resistant to MEM (MIC 4\u201332\u2009mg/L), TZC (MIC \u226564\u2009mg/L), GEN (MIC >32\u2009mg/L), and LEV (MIC \u226516\u2009mg/L), and susceptible to AMK (MIC 2\u20134\u2009mg/L). For MVB, two isolates were susceptible (MIC 2\u20134\u2009mg/mL) and two were resistant (MIC 32\u201364\u2009mg/mL).There were 4 patients with OXA-48\u2013producing isolates at baseline in the CAZ-AVI treatment group in the following Phase 3 studies conducted from 2012 to 2016: 1 from RECLAIM (cIAI), 2 from REPRISE , and 1 from RECAPTURE (cUTI). All 4 patients had Enterobacterales isolates collected from CAZ-AVI Phase 3 clinical studies, MVB , CAZ-AVI , and MEM  exhibited similar levels of in vitro activity, followed by AMK , GEN , LEV , TZC , and ceftazidime , according to Clinical and Laboratory Standards Institute (CLSI) breakpoint interpretation  and 32 E. cloacae . The most common ESBL gene type was blaCTX-M . The CTX-M-15 encoding gene was detected in 26 isolates, including 6 from C. freundii complex and 20 from E. cloacae. The CTX-M-3 gene was observed in 4 C. freundii complexes and 23 E. cloacae. A total of 28 isolates carried the gene encoding OXA-1/30. This gene was mostly observed among isolates carrying other ESBLs, mainly CTX-M-15. Plasmid-encoded AmpC gene CMY-42 was noted in one E. coli isolate  was among the most active agents in vitro, when tested against 53 AmpC-overproducing P. aeruginosa isolates collected from Phase 3 clinical studies, followed by amikacin , meropenem-vaborbactam (MVB) , TZC , gentamicin , meropenem , levofloxacin , and ceftazidime , when CLSI and European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoint criteria were applied  in the CAZ-AVI and control groups of all 4 clinical trials. The clinical outcome evaluation carried out in this study included only those patients with the isolates present at baseline. A total of 40 patients in the CAZ-AVI group, including 26 patients with baseline AmpC-overproducing Enterobacterales, and 14 patients with baseline AmpC-overproducing P. aeruginosa, were pooled together from the Phase 3 clinical studies: RECLAIM, RECAPTURE, REPROVE, and REPRISE. Clinical cures in patients with baseline AmpC-overproducing P. aeruginosa were 85.7% (n\u2009=\u200912/14) in the CAZ-AVI group versus 75.0% (n\u2009=\u20099/12) in the control groups. Clinical cures in patients with baseline AmpC-overproducing Enterobacterales were 80.7% (n\u2009=\u200921/26) in the CAZ-AVI group versus 85.0% (n\u2009=\u200917/20) in the carbapenem control groups.Isolates included in the Enterobacterales and P. aeruginosa owing to inhibitory activity of avibactam toward carbapenemases, including OXA-48 and KPC  were susceptible to ceftolozane-tazobactam, with only 8.1% of ceftazidime-resistant isolates  reported as susceptible to ceftolozane-tazobactam  are a significant advance in the therapeutic armamentarium against multidrug-resistant Gram-negative pathogens. The CAZ-AVI combination has shown potent activity against carbapenemase-producing  and KPC , 6. More and KPC . While tzobactam . The expblaOXA-48 when combined with meropenem , which is consistent with observations that vaborbactam does not inhibit blaOXA-48. The difference in susceptibility can be attributed to a higher breakpoint set for MVB (susceptible \u22644\u2009mg/L) than that for MEM alone (susceptible \u22641\u2009mg/L). An efficacy study of human-simulated exposure of MVB and MEM in the neutropenic murine thigh infection model revealed that the activity of both drugs against blaOXA-48 carrying Enterobacterales was poor despite more than a third of isolates falling within the susceptible range per EUCAST and CLSI MIC interpretation criteria  to AMK. The activity of AMK against isolates carrying blaOXA-48 in various combinations with ESBLs and other \u03b2-lactamases decreased by approximately 2\u201310%. As the presence of carbapenems or \u03b2-lactamases is not expected to impact aminoglycoside susceptibility, aminoglycoside-modifying genes, 16S rRNA methylases, and other potential mechanisms remain to be investigated. This in vitro susceptibility study demonstrated that CAZ-AVI was the most active agent in vitro against OXA-48\u2013producing Enterobacterales carrying multiple \u03b2-lactamases compared with other antibiotics, including \u03b2-lactams, \u03b2-lactam\u2013\u03b2-lactamase inhibitor combinations, and aminoglycosides.Vaborbactam, a cyclic boronic acid BLI, exhibited no activity against isolates harboring eropenem . Previoucterales , 16. In ctamases ; howeverEnterobacterales, CAZ-AVI showed promising results, even in monotherapy, for the treatment of patients with severe infections due to OXA-48\u2013producing Enterobacterales and limited therapeutic options  in the CAZ-AVI group versus 75% (n\u2009=\u20099/12) in the carbapenem control groups. Clinical success rates in patients with baseline AmpC-overproducing Enterobacterales were 81% (n\u2009=\u200921/26) in the CAZ-AVI group versus 85% (n\u2009=\u200917/20) in carbapenem groups. Similar results were observed in a systematic review and meta-analysis of randomized controlled trials comparing the clinical efficacy of CAZ-AVI and carbapenems for the treatment of ESBL/AmpC-producing Enterobacterales; the clinical response for AmpC producers in CAZ-AVI and carbapenem arms was 80% (n\u2009=\u200932/40) and 88% (n\u2009=\u200937/42), respectively  and P. aeruginosa (n\u2009=\u200953) were collected from 4 CAZ-AVI clinical trials: RECLAIM , REPRISE , RECAPTURE , and REPROVE . AmpC-overproducing Enterobacterales included Enterobacter cloacae (n\u2009=\u200949), Citrobacter freundii complex (n\u2009=\u200914), Enterobacter aerogenes (n\u2009=\u20098), Escherichia coli (n\u2009=\u20095), and Serratia marcescens (n\u2009=\u20091). The isolates included in the in vitro susceptibility evaluation were collected at various time points, including baseline, TOC, and EOT in the CAZ-AVI group and in the comparator study treatment groups.Nonduplicate clinical isolates of AmpC-overproducing K. pneumoniae isolates were collected from CAZ-AVI clinical trials . Nonduplicate clinical isolates of OXA-48\u2013producing Enterobacterales (n\u2009=\u2009113) were collected from the 2018\u20132019 Antimicrobial Testing Leadership and Surveillance (ATLAS) global program by medical centers in 25 countries. The OXA-48\u2013producing isolates included K. pneumoniae (n\u2009=\u200965), Klebsiella oxytoca (n\u2009=\u20096), Klebsiella aerogenes (n\u2009=\u20095), E. cloacae (n\u2009=\u200917), Enterobacter kobei (n\u2009=\u20092), E. coli (n\u2009=\u200911), and others (n\u2009=\u20094).Four OXA-48\u2013producing \u03b2-Lactamase gene screening in AmpC-overproducing clinical trial isolates was conducted using either microarrays  and PCR or a combination of both as described previously \u201323. The \u2013Enterobacterales isolates were screened for genes encoding carbapenemases  as well as the presence of coharbored \u03b2-lactamase genes encoding TEM, SHV, CTX-M-1 group, CTX-M-2 group, CTX-M-8 group, CTX-M-9 group, CTX-M-25 group, ACC, ACT, CMY, and DHA using a combination of microarray and multiplex PCR assays, followed by amplification and sequencing of the full-length genes. The screening was conducted at the central reference laboratory .OXA-48\u2013producing In vitro susceptibility testing was performed by the broth microdilution (BMD) method, using a custom-made panel manufactured by ThermoFisher Inc.  consisting of CAZ-AVI, ceftazidime, meropenem (MEM), meropenem-vaborbactam (MVB), ceftolozane-tazobactam (TZC), gentamicin (GEN), levofloxacin (LEV), and amikacin (AMK).P. aeruginosa  procedures (document M07) . Avibactruginosa .Enterobacterales and baseline AmpC-overproducing P. aeruginosa, as well as baseline OXA-48\u2013producing Enterobacterales, treated with CAZ-AVI or carbapenem comparators. Because the primary endpoint and analysis populations for the Phase 3 studies differed depending on the study and regulatory authority (U.S. Food and Drug Administration or European Medicines Agency), the clinical response  at TOC visit in the microbiologically modified intention-to-treat (mMITT) population was selected for comparisons. Briefly, clinical cure was defined as complete resolution or substantial improvement of signs and symptoms of the infection, such that no further antibacterial therapy  was necessary. TOC was assessed 21 to 25\u2009days after randomization  or 28 to 35\u2009days after randomization (RECLAIM).In this study, the clinical response at TOC was reviewed in patients with baseline AmpC- overproducing"}
+{"text": "Multinomial logistic regression estimates showed that depression symptoms, anxiety symptoms, and insomnia were significant negative predictors, while daily physical exercise was a significantly positive predictor. We found an overall downward trend in bus drivers\u2019 burnout, particularly in the sub-dimension of personal accomplishment. Mentally healthier drivers and those who were usually exercising were more resilient to occupational stress and less likely to suffer burnout.This study aimed to characterize job burnout in longitudinal trajectories among bus drivers and examine the impact of variables related to job burnout for trajectories. A longitudinal study was conducted in 12,793 bus drivers in Guangdong province, China, at 3-year follow-up assessments. Growth mixture modeling (GMM) was used to estimate latent classes of burnout trajectories and multinomial logistic regression models were applied to predict membership in the trajectory classes. In general, there was a decrease in job burnout in 3 years . Among those sub-dimensions, reduced personal accomplishment accounted for the largest proportion. GMM analysis identified five trajectory groups: (1) moderate-decreased ( Stress at work is an increasingly common phenomenon in society. Over a period of time, this stress may result in exhaustion, named burnout syndrome, which has been recognized as an occupational health problem . PreviouBurnout is a long-term response to chronic emotional and interpersonal stress at work and is defined by three dimensions: exhaustion, cynicism, and inefficacy . The dimThere are various theory models of the formation and development of burnout. Some models focus on the relationship between the three dimensions of burnout, which are usually described as successive stages . ExhaustIn addition, there are theories that focus on resources. For example, the Job Demands-Resources (JD-R) model suggests that when an individual\u2019s job demands are increasing, but there are not enough resources to meet and cope with these demands, this is when the burnout happens . Bus driPrevious research has found gender ,13, age In a study of professional drivers in China, job burnout was positively associated with aggression, which may be related to long working hours, poor working conditions, lack of discipline, and even passenger verbal and physical aggression . AlthougThe aim of this study was to identify and characterize potential job burnout trajectories and the relationship between job burnout and other related variables. By using the growth mixture modeling (GMM), we were able to offer a long-term understanding of job burnout for bus drivers and to provide evidence to develop better occupational mental health strategies.The study was a repeated measurement study performed at three time points. The three examined times were T1: 10 August to 9 October 2019; T2: 9 October to 9 December 2020; T3: 21 October to 9 December 2021. Bus drivers in Guangdong province, China, were invited to participate through online questionnaires using Sojump , marital status , the number of children, financial situation , body disease , mental illness , driving years (How many years have you been in the business?), working hours (How many hours do you work per day?), and daily physical exercise  were determined. More information can be seen in The Chinese version of Maslach Burnout Inventory-General Survey (MBI-GS) was used to measured burnout among bus drivers . The MBIThe Chinese version of the Patient Health Questionnaire (PHQ-9) was used to measure depression symptoms. PHQ-9 measures depression symptoms within two weeks . The totThe Chinese Generalized Anxiety Disorder Scale (GAD-7) was used to measure anxiety symptoms . The totWe used the Insomnia Severity Index (ISI) for evaluation of bus drivers\u2019 insomnia . The ISIThe Chinese version of the Buss\u2013Perry Attack Scale was used to measure aggression among drivers . It rangLatent class growth analyses were analyzed using Mplus, version 8.0. GMM analysis was an iterative procedure. It starts from a one-class model, and then successive models extract additional classes. After analyzing the one-class model, a two-class model was examined, followed by a three-class model, and so on. Model solutions were evaluated according to the Akaike Information Criteria (AIC), Bayesian Information Criterion (BIC), Entropy, posterior class probabilities, and the Adjusted Lo\u2013Mendell\u2013Rubin (LMR) test. Lower BIC and AIC values, as well as higher Log-Likelihood values, indicate a better model fit. The model\u2019s classification accuracy was summarized using entropy, which ranged from 0 to 1 and more than 0.8 meant that the classification quality of the model was acceptable. Based on the best-fit statistics of the model, the model was selected with the optimal number of classes. After selecting the best fit model, predictors of latent class membership were examined.A multinomial logistic regression analysis was conducted using Mplus 8.0 in order to identify predictors that distinguished the different trajectories, with class trajectories as dependent variables. The prediction variables were as follows: age, gender, marital status, number of children, financial situation, mental illness, driving years, working hours, daily physical exercise, depression symptoms, anxiety, insomnia, and aggression. Among these independent variables, age, the number of children, driving years, and aggression were continuous variables, and all other independent variables were coded as dichotomous variables. The Code of Latent class growth and multinomial logistic regression are visible in As shown in p < 0.00) and also a decline in reduced personal accomplishment , p < 0.00), but little change was observed in emotional exhaustion , p < 0.00) and in cynicism , p < 0.00).Because each dimension had a different number of items , we used the mean score of each dimension for comparison. As shown in The average latent class probabilities for most likely latent class membership is presented in n = 5062) was characterized by a low slope  and a low intercept  compared to others. Though the slope in class 2 was significant, the absolute value was close to 0. Therefore, class 2 was termed \u201clow-stable\u201d. The same as \u201clow-stable\u201d, class 5 (n = 3216) had a stable slope  while class 5 had a higher intercept  compared to \u201clow-stable\u201d. Therefore, class 5 was termed \u201chigh-stable\u201d. The third largest class  also had a high intercept  and a stable slope , so we named it \u201cmoderate-decreased\u201d. Compared to \u201cmoderate-decreased\u201d, class 3 (n = 141) had a more oblique slope  and had the highest intercept , so class 1 was termed \u201crapid- decreased\u201d. The last one was class 4 (n = 1504) and was characterized by an increased slope , therefore termed \u201cmoderate-increased\u201d.The largest class  the moderate-decreased, (2) the low-stable, (3) the rapid-decreased, (4) the moderate-increased, and (5) the high-stable.Results of the multinomial logistic regression were shown in \u201cModerate-increased\u201d and \u201clow-stable\u201d had comparable scores of MBI at T1, but one score became higher, and one score remained stable. We were curious about the potential predictors for this, so we set \u201clow-stable\u201d as the reference class. Similarly, we also set \u201cmoderate-decreased\u201d as the reference class to find the potential factors that \u201cmoderate-decreased\u201d and \u201chigh-stable\u201d trends were different. \u201cLow-stable\u201d and \u201chigh-stable\u201d were both stable, but we were curious as to why one scored high and one scored low, so we set \u201clow-stable\u201d as the reference class. In the two trajectories, \u201cmoderate-increased\u201d and \u201cmoderate-decreased\u201d, one was increased and one decreased. We were also curious about what their underlying factors were, so we set \u201cmoderate-decreased\u201d as the reference class.The overall results for the multinomial logistic regression showed that daily physical exercise, depression symptoms, anxiety symptoms, and insomnia were significant predictors. Daily physical exercise for more than 30 min was a protective factor, while depression, anxiety, and insomnia were risk factors.In addition to these factors , older drivers or those with more years of driving experience were more likely to have a moderate increase in MBI than those who had been stable-low. Drivers with lower than normal economic status or higher aggressiveness were more likely to be on a stable-high trajectory than a moderate-decreased trajectory. Lower financial situation, aggression, and longer driving years predicted a high stable trajectory in contrast to a stable low trajectory, while number of children predicted a stable low trajectory in contrast to a high stable trajectory. Older, more children, lower financial situation, and longer daily working time predicted a moderate increase in MBI compared to a moderate decrease.In general, job burnout of bus drivers tended to decrease over time, of which the most considerable component was the decrease in reduced personal accomplishment. The second component was cynicism, and the third was emotional exhaustion. It showed that the driver\u2019s burnout was mainly reflected in personal accomplishment, suggesting that it was effective to enhance personal accomplishment to reduce driver burnout. These findings were consistent with the observation in previous studies in which the professional efficacy dimension was a more central symptom in burnout syndrome . In the n = 141) was relatively small while \u201chigh-stable\u201d (n = 3216), \u201clow-stable\u201d (n = 5062). We would like to interview \u201crapid-decreased\u201d as a next step. This finding indicated that most bus drivers displayed a stable low in job burnout. Our data were similar to the result from Benjamin B.D\u2019s study in which burnout was relatively stable for employees without company changes [Based on the findings from this research, bus drivers were likely to observe five patterns of job burnout: (1) the moderate-decreased group, (2) the low-stable group, (3) the rapid-decreased group, (4) the moderate-increased group, and (5) the high-stable group. The largest group  had stable-low levels of job burnout and changed very little over the 26 months. The second group was class 5, which had stable-high levels of job burnout over the 26 months. Compared with class 2, class 5 tended to be stable-high while class 2 was more likely to be stable-low, although both class 5 and 3 had similar membership . Class 5 had stable-high levels of job burnout over the 26 months, and class 3 tended to be decreased moderately-. Compared with class 1, class 3 (1%) showed a rapid decrease. It was undeniable that \u201crapid-decreased\u201d was very researchable for reducing driver\u2019s burnout. However, in the regression, the results of the regression might be biased if compare \u201crapid-decreased\u201d with other groups, because the number of \u201crapid decrease\u201d  were lower or more stable for respondents who did daily physical exercise for more than 30 min. This was consistent with a systematic review and meta-analysis showing physical activity was beneficial for burnout syndromes ,38 and vJob burnout was associated with mental health. In our study, depression, anxiety, and insomnia were important predictors of driver burnout. This indicates that mentally healthier people were more resilient to occupational stress and less likely to suffer burnout. Similarly, a meta-analysis showed a significant association between burnout and anxiety and depression . In a meIn addition to the factors above, several demographics and burnout-related factors were also related to changes in job burnout. As for the number of children, if the driver was changing , burnout was changing, and the slope was steep. Having more children was associated with worse burnout scores. If the driver was in a stable state  and the slope of burnout did not change, drivers with more children were more likely to be in a peaceful state in their careers. In combination with age, the older the driver and the more children compared to rapid-decrease and moderate-decreased, the more likely the driver was in a precarious state. Age was unlikely to be a risk factor for low-stable, because the odds ratio (OR) was only 1.01. Low odds ratio meant that the effect of age was minimal and insignificant.Regarding the variables of drivers\u2019 financial status, financial status was a risk factor for driver burnout, except for drivers who already have been in low burnout. This suggested that company managers could try to increase burnout pay and invite experts to give lectures on financial literacy. Financial literacy was overlooked as a solution for burnout. Improving financial literacy was the first step in helping tackle financial problems that might worsen burnout . On the Moreover, although it has been reported that psychophysiological disorders were positive predictors of exhaustion and cynicism , this isThis article had the following limitations. Variables such as number of children, financial status, and driving years were discussed, but in terms of the comprehensiveness of the variables in JD-R, our study did not cover the variables involved in those theories well enough to argue for it, which is a shortcoming of our article. Regarding the JD-R, the individual\u2019s resources might include family functioning, social support, emotion regulation, etc., but in this study, none of these variables were measured. Furthermore, we focused more on negative variables and we covered fewer positive variables such as social support or quality of leadership. Factors related to driver personality and job characteristics have predictive effects on burnout, but this part of the study lacked exploration. Moreover, burnout, depression symptoms, anxiety symptoms and insomnia were examined through self-reported questionnaires; thus, participants\u2019 answers could be affected by potential self-report biases. In addition, we did not control for confounding factors between measurements, such as the COVID-19 epidemic, the gradual boom of the subway resulting in lower bus trips, bus accidents in the news during this period, etc. This study also ignored group heterogeneity, such as whether they worked night shifts, different driving routes, etc.In the 26-month longitudinal research, we found evidence that bus drivers\u2019 burnout showed an overall downward trend, mainly reflected in personal accomplishment. Public transportation companies could make certain interventions in this area to improve drivers\u2019 sense of accomplishment in a targeted manner, thus alleviating burnout. Mentally healthier drivers and those who were usually exercising were more resilient to occupational stress and less likely to suffer burnout. These results have practical implications. Public transportation companies could intervene to alleviate burnout by addressing the above factors. Future research could select subjects from the group for targeted interviews."}
+{"text": "Teachers' attitudes, behavior, and practices play an integral role in enhancing the students' learning activities. Teachers' strategies ensure the individuals' professional development by creating a healthy learning environment. The study's primary objective is to analyze students' attitudes toward learning English as a foreign language. The data was collected from the 359 colleges and universities students by adopting a convenient sampling technique. The study shows English self-concept and teacher behavior student orientation significantly positively impact attitudes toward learning English as a foreign language. Motivation for English and English Intentions mediate the relationship between English Self-concept, teacher behavior-student orientation, and Attitude toward learning English as a Foreign Language. The study motivates future studies to focus on the EFL, individuals' learning motivation and intentions in other domains across diverse contexts. In the globalized world of multilingual societies, the global education system has caused English to extend its spectrum from a common language to international reach. Today, English, holding a prominent linguistic position, has significantly become a vital spread serving worldwide communities. Its growing popularity has made it an imperial tool to foster global learning activities, substantially becoming a paramount requirement of today's academia . In partIn education, the students face considerable challenges in adjusting to the increasing demand of the ill-structured learning environment. These emerging difficulties affect the students' mental health and wellbeing. Given the articulation, the study shows that interest in students' mental health and wellbeing has grown exponentially in recent years . A healtIndeed, this matter of students' health and wellbeing holds a profound prominence in fostering the students' learning. Undoubtedly, today, English has gained massive foreign popularity. Its growth in education has made it function as an international tool for fostering learning outcomes . LearninIn language acquisition, an individual's success largely depends on their self-concept. Self-concept is a dynamic system of an individual's learned beliefs and actions . The selAlso, the study shows that the teachers' positive behavior significantly affects the students' academic success, learning intention, and achievement . TeacherSignificantly, besides teachers' having responsibility for influencing the students learning activity, student learning motivation also works as a critical factor in ensuring successful learning. Motivation is a primary source affecting the students learning. Given the illustration, the research shows that the lack of motivation makes the students quit learning English . PerhapsIndeed, today, the power to master the English language has gained considerable academic attention . PresentGiven the evidence, the literature showed that a high motivation boosts the students' intention to learn new concepts. However, besides this favorable relation, the study revealed that motivation to learn English as a foreign language has constantly faced backlash from the students . Indeed,However, against this drawback, the main objective of the present study is to investigate the factors stimulating the students' attitude toward learning English as a foreign language. The study expects that studying these factors will lead the students to embrace English learning as an international language fostering self-concept and teachers' behavior student orientation. Moreover, the study also highlights the mediating effect of motivation and English intention against EFL .Therefore, this incorporation appears to be a potential benefit in advancing the education sector. These insights develop a valuable contribution to the world's education sectors . In particular, the existing research gap showed that interest in the factors such as motivation and intention call for further investigation in the context of learning the language. Significantly, the study holds immense importance by empirically exploring the trajectories influencing students' perception of EFL learning. However, irrespective of specific influence, the unique research incorporated combinations of factors, thus elevating positive learning outcomes.Significantly, this knowledge of English proficiency provides distinctive characteristics to students by making them essentially advance their careers through effective English learning. The study offers the learners a comparative opportunity to experience EFL learning. It suggests improving the English learning motivation, self-concept, and intention to learn English to gain effective learning outcomes. Furthermore, the research findings also allow the educationists and government to understand the benefit of learning the English language, thus optimally supporting the English teachings. In particular, the research findings suggest adopting high linguistics teaching skills, thereby improving motivation and attitude.As a quick reminder: this current paper has split into six different sections where the proceeding section highlights the research background. Similarly, Section Methodology elaborates on the research methodology, and section Results and discussion documents the research outcomes. Lastly, section Conclusion discusses the study outcomes, with section Conclusion concluding the research paper.English, a global language, enhances the learners' fluency by significantly advancing their learning activity . Today, Individuals' success largely depends on the mastery of the English language, where positive self-concept plays an integral role in influencing the individual's learning attitude. The students' learning perception  helps them assess their need for EFL. In support, the study shows that the language self-concept provides feedback to the students, fundamentally shifting their focus toward EFL . The posH1: English Self-concept has a positive and significant impact on Attitude toward learning English as a Foreign Language.With the changing paradigm of global education, exploring teacher behavior has become an independent area demanding the establishment of English language teachings. The EFL has highlighted the teachers' role in advancing foreign language teachings from local to international settings. In support, the study from Turkey showed that teachers adopting novel teaching programs had significantly addressed the students' EFL learning needs . PerhapsStudents' learning depends on the teachers' teaching methods, style, and practices. Significantly, the instructors teaching material act as a guide in assisting the students learning activities. In this regard, today, teacher behavior- student orientation is gaining growing attention in shaping the students' attitude toward their studies. Teachers' teaching strategies fundamentally work to improve the students learning process. Accordingly, in support, the study shows that teachers' English learning strategies have improved the proficiency of the technical learners, thus directing their focus toward English learning .Altogether, teachers have a responsibility to influence student learning orientation. The teacher's positive emotion toward EFL elevates the need for an effective curriculum to strengthen the students' skillset. Teachers' constructive feedback and assignments improve students' understanding of English subjects, thus gaining academic success. The study shows that teachers providing the necessary learning materials develop an EFL environment  , substanH2: Teacher Behavior Student Orientation has a positive and significant impact on Attitude toward learning English as a Foreign Language.In the rapidly evolving learning climate, a positive self-concept boosts the students' confidence in the work activity. The self-concept motivates individuals to achieve good academic grades at all educational levels. English is the most important foreign language that has become the core for accelerating learning activities. Learning English requires getting knowledge about the subject. However, today, the motivation to learn English as a foreign language demands global attention. The self-concept makes the students identify the reason behind studying the English language, ultimately boosting their learning motivation .Indeed, academic self-concept predicts the students' motivation for learning. Given the illustration, the study shows that Self-concept increases individuals' autonomy, competence, and motivation . UndoubtH3: English Self-concept positively and significantly impacts motivation for English.The EFL demands the teachers' attention, involvement, and guidance in executing the learning activity. The teacher-behavioral-student orientation regulates the individuals' goal , fostering their desire to gain language proficiency . In the Undoubtedly, today, instructors widely contribute to students' learning process. The teachers provided curriculum helps the students to understand the challenging concept. English is a complex language that demands teachers' assistance. Teachers' behavior-student orientation allows teachers to make learning simple and easy with their teaching skills. Based on this statement, the research shows that teachers' teaching boosts students learning intention, thereby fostering the process of learning a language . Verily,H4: Teacher Behavior Student Orientation positively and significantly impacts English Intentions.Most students learn English because they believe it plays a paramount role in advancing their educations careers. However, without understanding the language's significance, students lose hope in learning the language. Undoubtedly, the English language has received global popularity by significantly making multinational students realize its benefit. A positive self-concept is a critical component of students' achievements and success. Given the illustration, the study shows that by recognizing the value of a foreign language, the students of Arab countries have significantly shifted their attitude toward learning English as a foreign language .The academic self-concept influences one's interest in the subject. In recent years, this idea fundamentally works well in education, especially in English. English self-concept makes students learn new vocabulary, thus performing well in every sphere of life. Given the illustration, the study from Vietnam showed that students' perception influences their career interest in EFL, thereby increasing their career intention in English . ConsistH5: English Self-concept has a positive and significant impact on English Intentions.Motivation as the instrumental factor acknowledges the need for acquiring English as a foreign language. Teachers' behavior- student orientation, an integrative approach, makes the students study English with more motivation and enthusiasm, thus influencing their academic choices. Motivation to learn English as an international language is highly related to teaching aspects  . Today'sH6: Teacher Behavior Student Orientation positively and significantly impacts Motivation for English.Motivation develops a strong bond with students' learning attitudes. Academic progress highly depends on students learning motivation and their interest in language. The students come to the classroom to learn something new. However, understanding these motives increases their morale by realizing the worth of studying the language. Generally speaking, motivation plays an integral role in advancing English learning as a foreign language. English boosts the students' motivation and learning attitude. Given the explanation, the study indicates a positive relationship between student motivation, learning attitude, and EFL .The acquisition of English supports both the teaching and learning orientation. Perhaps, learning this language demands individuals' time, attitude, and motivation. Aptitude and motivation are effective drivers of developing language skills . Indeed,In the same vein, motivation leads the individual to gain language proficiency. It helps the students to raise their competency to an expert level. Accordingly, the study shows that the significant shift in language learning has made the teachers' characteristics increase the students' participation , thus enH7: Motivation for English has a positive and significant impact on Attitude toward learning English as a Foreign Language.H7(a): Motivation for English mediate the relationship between Self-concept of English and Attitude toward the learning of English as a Foreign Language.H7(b): Teacher Behavior- Student Orientation mediates the relationship between English Self-concept and Attitude toward the learning of English as a Foreign Language.Fundamentally, English as a medium of language has grown over time, thus fostering the students' professional activities. The EFL inspires the students to learn the language at all school levels. It is an instrumental motivator encouraging the students to involve in EFL learning. Given the explanation, the study shows that EFL positively influences the students' attitudes toward English . The stuMoreover, the study also shows that EFL motivates the students to obtain positive learning outcomes by strongly shifting their focus toward English . The selAdditionally, by understanding the language learning belief, teachers provide instrumental support to their students' by arranging sufficient teaching materials for their assistance . However, the strength of the educational system cannot be judged without the EFL. Therefore, in coping with the growing educational requirement, the study showed that innovative teaching methods have rigorously improved the students' English reading and writing skills, thus making them confident in the learning methods , 60. HenH8: English Intentions have a positive and significant impact on Attitude toward learning English as a foreign language.H8(a): English Intentions mediate the relationship between English self-concept and attitude toward the learning of English as a foreign language.H8(b): English Intentions mediate the relationship between teacher behavioral student orientation and attitude toward the learning of English as a foreign language.The study used quantitative research to examine the impact of English self-concept, and Teacher behavior student orientation on the attitudes toward learning English as a foreign language. Additionally, the study examines the mediating effect of motivation for English and English intentions between English self-concept, teacher behavior and attitudes toward learning English as a foreign language. The study utilized the positivism philosophy, which means data gained by scientific observations and measurements. The deductive reasoning approach is applied in this study. The data was collected from the students studying in the Colleges and Universities of China. The study adopted the convenient sampling technique for the data collection process. Previously developed structured questionnaires were used for data collection.Moreover, the five Likert scales were used to measure the survey, ensuring the reliability and validity of items. However, before sending the questionnaires to respondents, a cover letter is attached to ensure the confidentiality of respondents for research objectives. The sample of 430 questionnaires was distributed to the students at different colleges and universities in China, and data was collected electronically. After that, 400 questionnaires were received, and finally, 359 questionnaires were chosen to analyze the data with an 89% response rate. The study used the smart PLS for the Partial least square's structural equation model.Common method bias using Harman's single-factor approach was applied in this study. The variance extracted by one single factor is 11.961% which is <50%, indicating no common method bias .Reliability, validity, and discriminant validity were analyzed in the measurement model. In reliability, the Alpha and CR values must be investigated, and both must be higher than 0.6 , 64. ConAs shown in This research also checks the VIF values for English intentions, motivation for English, and learning English as a foreign language. English self-concept and teacher behavior student orientation. All values fall below the threshold see .This research applied the PLS-SEM technique using Smart-PLS software version 3.3.3 for hypothesis testing . The booOn Attitude toward learning English as a Foreign Language was accepted and confirmed with a standardized path coefficient of 0.222. The impact of English Self-concept on English Self-concept is positively significant at the 0.001 level. H2) Teacher behavior-student orientation positively impacts attitude toward learning English as a Foreign Language was accepted and confirmed with a standardized path coefficient of 0.194. The impact of English Teacher behavior student orientation on Attitude toward learning English as a Foreign Language is positively significant at the 0.001 level. H3) English self-concept positively and significantly impacts motivation for English was accepted and confirmed with a standardized path coefficient of 0.423. The impact of English self-concept on motivation for English is positively significant at the 0.001 level. H4) Teacher behavior student orientation has a positive and significant impact on English intentions was accepted and confirmed with a standardized path coefficient of 0.335. The impact of Teacher behavior student orientation on English intentions is positively significant at the 0.001 level. H5) English self-concept positively and significantly impacts English intentions was accepted and confirmed with a standardized path coefficient of 0.411. The impact of English self-concept on English intentions is positively significant at the 0.001 level.H6) Teacher behavior student orientation has a positive and significant impact on motivation for English was accepted and confirmed with a standardized path coefficient of 0.347. The impact of Teacher behavior- student orientation on motivation for English is positively significant at the 0.001 level.H7) Motivation for English has a positive and significant impact on attitude toward learning English as a foreign language was accepted with a standardized path coefficient of 0.271. The impact of motivation for English on attitude toward learning English as a foreign language is positively significant at the 0.001 level.H8) English Intentions has a positive and significant impact on Attitude toward learning English as a Foreign Language was accepted with a standardized path coefficient of 0.279. The impact of English Intentions on attitude toward learning English as a foreign language is positively significant at the 0.001 level.p-value was <0.05. H7b) Teacher behavior student orientation mediates the relationship between English Self-concept and attitude toward the learning of English as a foreign language was accepted at the beta value of 0.0.094, and the p-value was <0.05. Hypothesis H8(a) and H8(b) were accepted and confirmed at the beta value 0.115 and 0.093, respectively (as shown in H7a) Motivation for English mediates the relationship between Self-concept English and attitude toward the learning of English as a Foreign Language was accepted at the beta value of 0.114, and the Today, English has become a dominant language used worldwide. It is spoken and understood by the majority of people. It has made cross-culture education possible for the global communities . In particular, learning the English language has become a necessity in academics. Its increasing significance has made today's educators well aware of English teaching, concepts, and benefits. Indeed, in understanding its widespread, this study presents a generalized model illustrating the students' intention, motivation, and teachers' student orientation to influence the EFL process. Fundamentally, the study explores the factors that alter students' English language learning attitudes. Accordingly, section Conclusion provides a complete understanding of the benefit of EFL, thus significantly predicting students' intention to learn English as a foreign language. Altogether, this section highlights the study results compared to the previous works.Undoubtedly, people have different reasons to learn the English language, but learning English as a foreign language demands a positive student attitude. Fundamentally, today's educators encourage the students to self-control their learning activity by regulating their skillsets and abilities. Therefore, for learning to be more effective, self-concept has drastically been adopted by the worldwide knowledge members . Education in all stages embraces the self-concept process as a beneficial instrument for enhancing students' English learning. Given the articulation, the study shows that self-concept allows the students to foster their EFL attitude and proficiency . Also, fVerily, learning English as a foreign language has become essential for students. English exposure is crucial for a learner's self-concept. In this regard, the study shows that self-concept makes students realize the value of learning the English language, thus motivating them to indulge in the English learning process . Also, tAccordingly, positive teaching is a critical driving force that nurtures the classroom environment. Teachers play an instrumental role in elevating the students' motivation in the English teachings. Students' motivations in English as a foreign language influence students' learning process. Given the illustration, the teachers' support increases the students' autonomy and motivation for language learning in the future . StudentSimilarly, students' language learning intentions, attitudes, beliefs, and abilities foster the classroom environment. Hence, understanding this phenomenon makes the educationists predict the students' foreign language learning intentions. In the illustration, the study shows that students prefer to take foreign language learning classes over the other classes . Indeed,In recent years, the global education sector has embraced the English language as a profound learning tool for increasing the students' motivation and intention. Apart from one's native language, English is the dominant learning of this modern world that has inevitably gained acceleration in the international world. English as a foreign language has brought different cultures together, thus encouraging the countries to mark it as a fundamental tool fostering the students' education. However, besides its increasing significance, English is a popular topic that has remained unaccepted in education.Accordingly, the main objective of the present study was to investigate the factors stimulating the students' attitude toward learning English as a foreign language. These factors will lead the students to embrace English learning as an international language fostering self-concept and teachers' behavior student orientation. Moreover, the study also highlights the positive mediating effect of motivation and English intention against EFL . In particular, the existing research gap showed that interest in the factors such as motivation and intention. Significantly, this study holds immense importance by empirically exploring the trajectories that influence students' perception of EFL learning.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. The patients/participants provided their written informed consent to participate in this study.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "This study aims to examine and compare female and male Kurdish EFL students\u2019 level and type of motivation based on L2 motivational self-system components and to identify their dominant type of motivation. The participants of this study were 118 students  were randomly selected as the participants of this study from different cities of Erbil governorate in Kurdistan region of Iraq. A Learners questionnaire used following the application of semi-structured interview sessions with learners who participated in the study. The data obtained from the questionnaire were analyzed through SPSS software, and the interview results were interpreted and analyzed through content analysis. The findings of this study revealed that both male and female Kurdish EFL students generally have high levels of motivation and the attitudes of both groups of learners were found to be generally positive towards English language and its learning. The most frequent motivational strategies, ways of motivating and arranging for them a suitable culture and recommended by the teachers in this study are helpful, especially for teacher educators in teacher training programs, so that they can get aware of the most used strategies by Kurdish EFL teachers in high school to be the leadership for this purpose, and try to improve and correct them, and try to teach them the most effective strategies and ways of raising motivation in the students. Every human being is born with the learning ability and the learning happens in every stage of our lives consciously or unconsciously according to our needs. However, considering the educational programs, like any other formal or specific programs, learning happens consciously and therefore it cannot effectively and successfully takes place if the students do not have enough motivation. In fact, conducting research on the issue of motivation and its related concepts can help us to find the answer to the question of why some students are eagerly involved the class activities, do their homework eagerly, and do much effort in learning, while some others avoid doing their class assignments and if they do the class activities or study, they do it reluctantly. More specifically, in the case of foreign language learning, the students need to find enough reasons and interest for learning the target language. Since their beliefs and attitudes towards learning a foreign language, and in general the extent of their motivation, is determiner of their language learning achievement, it is important to study about their level of language learning motivation, their dominant type of motivation, and the factors which play the most important role in motivating them. Accordingly, this study aims at investigating and comparing the extent of male and female Kurdish students\u2019 motivation in learning the English language as a foreign language in their classes, to explore the relationship between L2 Motivational Self System components  with their language learning motivation and their language proficiency; and also to identify the most frequent type of motivation they have in terms of integrative and instrumental, and their attitudes towards language learning in general. Accordingly, considering the theoretical framework, this study has been partly conducted based on the L2 motivational self-system model proposed by Conducting this study is very beneficial in providing useful information for the educational decision makers, authorities, teachers, and scholars in this field to answer such questions and look for appropriate strategies and plan effective programs to enhance the students\u2019 learning motivation and in the case of our study, the students\u2019 language learning motivation and attitudes.While many studies have been conducted in the area of language learning motivation, these types of studies on the Kurdish students have been rarely found in the literature, which calls for more investigation in this context.To date, many research works have been conducted in the domain of motivation and foreign language learning, most of which were carried out in recent years e.g., , 2012, hEvery research work comes with some limitations and similarly, this study also faced with few of them. One of the limitations that were faced in this research was due to some students\u2019 unwillingness to participate in the study. In this regard, some of the students did not fill the questionnaires that were administered to them and did not turn them back and in case of interview sessions, majority of the participants did not permit the sound-recording, which could be helpful in giving more complete and accurate data. It must be declared that the data were collected during the limited time that the schools were opened in Kurdistan region of Iraq before they were closed again due to the COVID-19 virus pandemic, and all ethical considerations were observed during the data collection process.Considering the type of motivation that students have regarding English language learning, Considering the learners\u2019 motivation, in the studies by Regarding the factors affecting students\u2019 motivation, Regarding EFL students\u2019 perceptions towards English language learning and the issues related to the foreign language learning , we can briefly mention to the following studies and their findings that correspond to the findings of the present study:Regarding the L2 motivational self-system, The results of the study by The results of the study by In contrast to the above findings, in the studies by Generally, this study intends to answer the following 2 main questions:Is there any difference between male and female Kurdish students\u2019 beliefs and perceptions towards their own motivational level, type of motivation, and the effectiveness of the motivational strategies used by their teachers?Is there any difference between the L2 motivational system components  which mostly affect male and female Kurdish students\u2019 motivation and language proficiency?The participants of this study were 118 students, including 46 female and 72 male Kurdish students. The detailed description of the participants is illustrated in Moreover, 36 students who showed consent and willingness to be interviewed were selected to participate in the interview sessions in order to elaborate on their thoughts and perceptions.As a quantitative data collection tool, a contextually adapted and modified version of a questionnaire that was originally developed by In designing this questionnaire, a 6-point Likert scale was used for the items, where considering the statement-type items, the participants were supposed to choose a number for their answers from 1 to 6 .In this study, the qualitative data obtained from interview sessions for learners were analyzed descriptively through content analysis method. The researcher partly designed the questions raised in the interview sessions according to the aims of the study, and the other part was based upon the items they had responded to before in the questionnaires. In this regard, the participants\u2019 responses were typed exactly the same way as they were uttered, so that the data would be kept authentic and without any modification or correction. Then all the participants\u2019 answers were typed and listed under each question that was asked to organize the data and to facilitate the data interpretation process. After that, the answers were all reviewed by the researcher through document and content analysis, and then through open-coding strategy, the whole data were analyzed and interpreted.The learner motivation questionnaire was administered to the students at the end of the academic semester, so that the students could better answer the questionnaire for they had experienced the English language class for enough time with their teacher. The questionnaire was also administered in the presence of the researcher, so that in case of any ambiguity the researcher could answer the participants\u2019 questions and solve their possible problems in understanding the questionnaire. Moreover, the researcher translated the items which seemed difficult to understand for the students, so that they could answer without worries of not understanding, and so their responses would be more accurate.After collecting data through questionnaire, the volunteered participants were selected to be interviewed. The researcher took the notes of the students\u2019 responses for most of the participants did not agree their voice to be sound recorded. So the researcher also used an assistant in order not to miss anything from the participants\u2019 responses to the interview questions.Regarding the ethical considerations, before administrating the questionnaires, the nature and goals of the study were briefly explained to the students in a way that they do not affect the answers they provide and hence the reliability of the data which was going to be collected. The participants were also assured of the confidentiality of the data they would provide and they were asked to return the questionnaires anonymously.The same ethical issues were also considered in data collection procedure for the interview. Before conducting the interview, the participants\u2019 permission was asked to sound record the interview sessions to be used for the easier data analysis, and therefore for those who felt uncomfortable, the interview was conducted without sound-recording, and only by note-taking. Unfortunately, few of the participants agreed on sound recording the interviews, and the others were note-taken.It must be asserted that all participants were assured of the confidentiality of the data they provided and that the researcher is the only one who will have access to the collected data and specially the sound-recorded data.The participants\u2019 responses to the questionnaire were analyzed through the frequency analysis method which shows the distribution of the participant\u2019s responses to each item in the questionnaire and was interpreted based on each of these 7 mentioned scales or categories that this questionnaire aimed to assess. First, the results of the participant\u2019s responses will be represented as a whole for all seven scales. Then the responses of the participants in the groups of females and males will be represented separately and compared.To analyze the data collected through questionnaire, SPSS software was used and the results were represented through tables and diagrams, using frequency analysis and descriptive statistics. On the other hand, content analysis method and open-coding strategy was used to descriptively analyze and interpret the data obtained from interview.The participants\u2019 responses to the questionnaire were analyzed through the frequency analysis method which shows the distribution of the participants\u2019 responses to each item in the questionnaire, and were interpreted based on each of these 7 mentioned scales or categories that this questionnaire aimed to assess. First the results of the participants\u2019 responses will be represented as a whole for all seven scales. Then the responses of the participants in the groups of female and male will be represented separately and compared as it can be seen in According to According to the obtained results, as shown in According to the results of comparing the male and female students\u2019 responses to the questionnaire scales, as shown in Considering the results of learners\u2019 interviews and based on the factor of gender, it is noteworthy that generally no significant difference had been observed between male and female students\u2019 general beliefs and attitudes in the interview section. Hence, since most of the participants\u2019 responses were almost similar to each other, we raised a few examples of responses in elaborating on learners\u2019 responses to the interview questions.Learners\u2019 responses to question 1 (How much do you consider yourself motivated or demotivated regarding English language learning?)The majority of the participants (79.6%) considered themselves motivated in learning the English language, while the rest of them considered themselves demotivated in this respect. To mention some of the responses and the participants\u2019 elaboration on their answers we can refer to the following responses which belonged to the students who considered themselves motivated:\u201cMotivated. Yeah, I am motivated enough to eagerly learn this language. You know, I like to travel abroad and visit different countries and meet people from different cultures and of course languages. I feel learning about different languages is amazing, but I prefer English the most and I choose it to learn and I do it eagerly because I think no matter where to travel, wherever I go, there are people who can understand English and I can use it to communicate different people. That\u2019s why I feel very motivated in learning this language.\u201dLearners\u2019 responses to question 2 (What factors do you think were effective in motivating or demotivating you for learning the English language?)The most frequent effective factors that the participants mentioned, respectively, included teacher (90.6%), the better job opportunities in the future (82.8%), more future academic success (75%), educational setting (60.9%), class atmosphere (87.5%), learning facilities (70.3%), method or ways of teaching by teachers (93.7%).To mention some of the responses and the participants\u2019 elaboration on their answers we can refer to the following responses:\u201cI think my teacher played the most important role in demotivating me in learning English. I think teachers\u2019 teaching practices, strategies, and behaviors are very effective in motivating or demotivating the students. My teacher\u2019s way of teaching for example is not attractive and his behavior is so cold.\u201cI think many factors are involved including families, society, and teachers mostly, and at the end, school and educational setting. I think they all are important factors. They can all have an impact on the students\u2019 motivation or demotivation.\u201dLearners\u2019 responses to question 3 Most of the participants considered learning English very beneficial, especially for future work (84.3%) and continuing education (76.5). The other reasons that they mentioned for the benefits of learning this language included the ability of communication with people from all over the world (87.5%), social status and prestige that knowing this foreign language can give to a person in Erbil (57.8%), watching original movies in English (45.3%), and the last one was travelling and studying abroad (67.1%).To mention some of the responses and the participants\u2019 elaboration on their answers we can refer to the following responses:\u201cI think it\u2019s very useful especially because we live in a new world. English is the international language and if we know it we can use it everywhere, we can communicate, trade and study everywhere with different people from countries other than ours.\u201d\u201cOf course, it is important and beneficial. It is the international language and the main language of the world. We live in the era of technology. Working with computers, internet, and working with different programs mostly need knowing English and if we know it well it can be really helpful. It makes finding a good job easy and we can watch our favorite English language movies and read original books easily in English and do not need to look for their translations or subtitles of the films in our own language.\u201dLearners\u2019 responses to question 4 (What do you think about English language and its instruction at school?)Majority of the participants had positive attitudes towards English as a foreign and international language (96.8%), and most of them also considered it crucial to be included in school curriculum and be taught as one of main subjects (93.75). To mention some of the responses and the participants\u2019 elaboration on their answers we can refer to the following responses:\u201cI think English is a very sweet language, and its learning is easy if it is taught well at school.\u201d\u201cI feel learning English gives me self-confidence and power. I like to become fluent in English language speaking and use it easily. But I feel it\u2019s not being taught well at schools. For example, in our school English teachers mostly concentrate on grammar rather than conversation.\u201d\u201cEnglish language is not my field of interest, but I study this school subject a lot and I want to learn it well because it is one of our main school subjects and the score, I get for this class is important both for me and for my family.\u201dLearners\u2019 responses to question 5 (To what extent do you think your teacher was effective in motivating you?)The majority of the participants (9 3.7%) considered the teacher\u2019s role very effective in motivating them. To mention some of the responses and the participants\u2019 elaboration on their answers we can refer to the following responses:\u201cOur teacher has a great role in this respect. Especially due to the interesting class activities that he assigns to us and the out-of-class activities. For example, he asked us to search in the internet and collect information about a specific subject. Activities like making wall newspaper. His teaching practice and the way of his teaching is so interesting and attractive to me.\u201d\u201cI really like my English teacher. One of the major reasons for my motivation in learning English is my teacher. Before previous years, I was weak in the English language and I even felt that I did not like studying English at all. But this year, my English teacher made me attracted in learning this language. Especially with his specific behavior and the attention he paid to me in English class, he gave me self-confidence and raised my self-esteem. He always encouraged me and did not look at me as a weak student in class, that\u2019s why I became encouraged and motivated to study more, and now I like to study more to make my teacher and my family more satisfied with me.\u201dLearners\u2019 responses to question 6 [To what extent do you think the educational system was effective in motivating you ].Most of the students who participated in this study considered the role of the educational system very important and effective in motivating the students (98.4%). To mention some of the responses and the participants\u2019 elaboration on their answers we can refer to the following responses:\u201cI think what is being taught at school has been effective in creating motivation in me. For example, our English textbook is very good. Nearly all needed contents are included in this book. It is attractive to some extent. I think we have a good educational system because overall, school is the main motivating factor in me.\u201dThe answers to the research questions of this study are represented and discussed below in light of the obtained results that were elaborated in the previous section.Is there any difference between male and female Kurdish students\u2019 beliefs and perceptions towards their own motivational level, type of motivation, and the effectiveness of the motivational strategies used by their teachers?\u201d the results showed that there was no significant difference between male and female Kurdish students\u2019 beliefs and perceptions towards their own motivational level, type of motivation, and the effectiveness of the motivational strategies used by their teachers. Based on the findings of this study, majority of Kurdish students were interested in learning English language as a foreign language and they mostly considered themselves motivated in learning this language. They also mostly preferred to learn English for itself, not as a class requirement. It must be asserted that no significant difference was observed in the responses of the male and female students. we should refer to the results which showed that in general all Kurdish EFL learners in our study mostly had both instrumental and integrative orientations towards learning English. However, female students showed to be more instrumentally motivated than male students, while for male students, integrative orientation was much higher than being instrumentally motivated. They had integrative motivations due to the fact that they mostly loved learning English for they considered it important for facilitating the communication and interaction with foreigners. They mostly have strong desire to learn the target language and to become fluent in that. On the other hand, they showed to be instrumentally motivated since they were mostly motivated to learn foreign language for the reasons such as getting a good job and becoming more educated, and also due to the fear of getting bad mark or disappointing their family and culture.Considering the first main research question of this study \u201cWhat is the Kurdish EFL students\u2019 attitude towards English language and its instruction at school?,\u201d the results of this study showed that generally, Kurdish EFL students  had positive attitudes and perceptions towards learning English as a foreign language. In fact, they generally loved English language and had overall favorable attitudes regarding learning this language as a foreign and international language, and most of them considered it very important to be included in school curriculum and be taught as one of main school subjects.Regarding the research question \u201cThe other related findings also showed that both male and female Kurdish learners of English language had positive attitudes towards native English speakers and communicating with native speakers of English was interesting to them. Moreover, considering learners\u2019 attitude and beliefs about their English classes, most of Kurdish learners who participated in this study showed their love and interest regarding English subject, however they mostly declared that their class atmosphere was not enjoyable enough and it is somehow boring.Exploring the attitudes and beliefs of male and female Kurdish EFL students also showed that learning English is very beneficial to them, especially for their future work and continuing education, increasing the communication with people from all over the world, the social status and prestige that knowing this foreign language can give to a person in Erbil, watching original movies in English, and the travelling and studying abroad. Considering the role of teacher in motivating the students from the Kurdish EFL learners\u2019 perspective, it was found that while they  generally believed that teacher\u2019s role is very effective in motivating and demotivating the students, majority of these students did not find their own teachers\u2019 role and practice much effective. In fact, they mostly believed that their teachers were generally good and inspirational majority of Kurdish EFL students  considered educational system as being highly effective and believed that it plays a very important role in motivating the students in learning English language.Other findings of this study based on Kurdish EFL students\u2019 opinions on the related issues which are interesting and noteworthy indicate that Majority of Kurdish students believe their parents are good motivators and have special role in directing and encouraging them towards learning English as a foreign language, and they mostly considered their parents as being actually supportive enough in helping them with their studies and pleasing them is very important for the students. Moreover, better future job opportunities, more future academic success, educational setting, class atmosphere, teaching aids and learning facilities, teachers along with their behaviors and methods or ways of teaching, are found to be the most frequent factors which the Kurdish EFL students perceive as being effective in motivating or demotivating them.\u201cIs there any difference between the L2 motivational system components  which mostly affect male and female Kurdish students\u2019 motivation and language proficiency?\u201d based on the results obtained from the Kurdish EFL learners\u2019 responses to the L2MSS questionnaire, It was found that generally all three main components of L2 motivational self-system  highly affect both male and female students\u2019 motivation almost similarly and consequently affect their language proficiency. Considering other motivating factors that was assessed through questionnaire, it was also found that family and culture, positive attitude towards learning English, and criterion measure that represents students\u2019 extent of interest in English language class play a great role in motivating both female and male students. Both male and female students did not show much fear of assimilation, which is good, because having fear of assimilation might make the learners less motivated and make them show avoidance behavior. However, both male and female students did not consider the culture of their English language classes interesting and motivating, and both groups showed high range of English language anxiety, while the English anxiety of female students were slightly higher than male students. Therefore, special attention must be directed towards improving the motivating factors and removing demotivating factors by teachers and the education system.Concerning the second main research question All in all, considering the EFL students\u2019 level of motivation, the results of this study are in line with results of the studies by This study also provides support for the studies by The findings of this study regarding factors affecting the students\u2019 motivation are also consistent with the findings of Who put emphasize on the positive and most effective role of teachers on enhancing students language learning motivation.The findings of this study are also in line with the findings of Considering the effectiveness of L2 motivational self-system components and its relationship with students\u2019 motivation, the findings of this study provide support for the findings of the studies by The results of this study has provided many useful information about Kurdish students\u2019 extent of English language learning motivation and the factors that have impact in motivating and demotivating them by the declaration, experience and beliefs of the participants of this study. All of these information has implications for the teachers, policy makers, educators and all involved in the education system. In general, to provide better learning environment and try to find the best strategies and policies to enhance students\u2019 English language learning motivation and consequently increase their language learning achievement in general.The fact that in this study we found majority of Kurdish EFL students are highly motivated by having both instrumental and integrative motivation is very promising, which shows that education system in this region of Iraq can be hopeful about the future of their students and the effectiveness of their system of education in general. Moreover, teachers can take advantage of the finding which shows the students\u2019 integrative motivation is higher than their instrumental motivation, and try to choose and plan appropriate strategies and ways to help them also increasing their instrumental motivation.The most frequent motivational strategies and ways of motivating used and recommended by the teachers in this study are also helpful, especially for teacher educators in teacher training programs, so that they can get aware of the mostly used strategies by Kurdish EFL teachers, and try to improve and correct them, and try to teach them the most effective strategies and ways of raising motivation in the students.Generally, in the light of the main findings of the study, it can be concluded that special attention must be paid to the students\u2019 motivation for the important and crucial role that it plays not only on the students\u2019 educational achievements and success, but also on their future job opportunities and social status. At the end, it is hoped that the outcomes of this intensive research work be useful in developing the students\u2019 English language learning motivation and in helping the education system and the teachers in setting appropriate plans and strategies to make that happen.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/The studies involving human participants were reviewed and approved by Ethical Committee Board of Cyprus International University. The patients/participants provided their written informed consent to participate in this study.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "The emerging field of human\u2013computer interaction (HCI) opens up more opportunities for technology integration in language learning. Technology creates more workforces in the Science, Technology, Engineering, Mathematics (STEM) field, yet the number of STEM pursuers is declining due to poor command of the English language. There is a gap in English vocabulary for STEM, which needs a novel solution from the perspective of HCI. This study aims to explore the needs of STEM learners in creating an English language competency mobile module. The methodology used is through a qualitative study. Seven STEM learners, 17 years old, participated in semi-structured interviews. The results from the interview are divided into four main themes: (1) the importance of learning English, (2) problems of learners, (3) strategies of English language learning, and (4) learners' readiness in using a mobile app. This study is significant for mobile app designers, English language teachers, and course designers as the input will provide an overview of the STEM learners' needs in the English language. Plus, designing a mobile app from the learners' perspectives gives more effectiveness to HCI, rendering success to second language acquisition. Future work can design and develop a mobile app to enhance STEM learners' English language competency based on the learners' perspectives. Technology creates more workforces in the Science, Technology, Engineering, Mathematics (STEM) field. The STEM field is vast, yet the number of STEM pursuers is declining due to poor command of the English language among non-native speakers  was pioneered by Professor Ben Shneiderman and has since evolved with the industrial revolution, providing more fresh ideas for its integration  importance of English, (2) problems in learning English, (3) learning strategies used in learning English, and (4) mobile readiness in learning English. Only one researcher took part in this interview session to ensure that learners did not feel pressured by the presence of many researchers. The researcher also took field notes to note on participants' non-verbal behavior. Researchers did not use any template for the field notes.Data from the interview transcripts were analyzed using thematic analysis. The transcripts were analyzed using the Atlas.ti software. The process included six steps by Braun and Clarke . The proFrom the analysis, eleven subthemes emerged, categorized into four main themes: (1) Importance of English, (2) Problems in learning English, (3) Strategies in learning English, and (4) Mobile readiness in learning English. Why do you think English is important for you as a secondary school student?Question: in university, everything is in English.\u201d Participant 1 mentioned, \u201cfor me, learning English is easy if we start from young or maybe we keep learning and learning, like that.\u201d These perceptions showed that the participants had been exposed to learning English since they were young, perceiving that English is important for education.The researchers asked participants why English is essential to identify the learners' view of learning English. Across the responses, education is the most crucial reason for learning English. English is used in many subjects of education, especially Science and Mathematics. Also, in tertiary education, most STEM courses are taught in English, making the participants aware of the importance of English. The response from Participant 6 mentioned, \u201c, \u201cwe can talk with our idols\u201d (Participant 5). Another response mentioned, \u201ceasy to communicate with other people, can speak easily\u201d (Participant 4). These responses referred to the participants' perception of learning English to communicate with other countries. One participant said, \u201cwe will not be easily deceived by foreign people\u201d (Participant 5). These responses showed that learners in high school might have the thought of going abroad; hence learning English seemed to be important to them for communication.Based on the responses, a few participants mentioned that learning English for communication is essential, which many learners value. Five participants said that English is vital for communication. When prompted to, one participant mentionedEnglish is the language people always use. It's wide and wherever we go, other than our own language, Bahasa Melayu, we have to have side language. The most common is English\u201d (Participant 1). In enhancement, Participant 2 also quoted the word lingua franca in his response, \u201cThe English language is one of the lingua franca languages, and almost all country in the world know English as a communicating language\u2026 Even though English is not our mother tongue language, but we have to learn it because English is also one of the lingua franca languages\u201d (Participant 2). Additionally, Participant 1 added, \u201cand when we go anywhere, it's not only our language, but we have to learn English so that it's easy\u201d. Looking into these responses, the notion that English is an international language becomes one reason for learning English.Another importance of learning English is because English is an international language. One of the participants mentioned, \u201cWhat are the difficulties or challenges that you've faced in learning English?Question: they are weak in vocab and grammar. Vocab is important because we want to write essays and all that, we use vocabs\u201d (Participant 7). Another response noted, \u201cthe problem of grammar, grammar. Lack of vocabulary\u201d (Participant 3). Most of the responses revolved around grammar and vocabulary. The researcher followed up with prompt questions to know why they said grammar and vocabulary are difficult, and one response mentioned, \u201cbecause if we don't understand the vocab, we won't understand\u201d (Participant 1). Language related problems that learners face are related to grammar and vocabulary. They know that they would not acquire other language skills without vocabulary.The researcher then asked the second question to know the learners' problems in learning English. One of the problems was related to language difficulties. One response highlighted, \u201csometimes when they try to speak English, people will make fun of them, like judgmental society\u201d (Participant 6). Another response highlighted that \u201cdidn't get support from friends, maybe family\u201d (Participant 4). These participants felt anxious when they did not have the support from their surroundings to encourage them in using English. Another response gave an example of anxiety that he felt when he spoke English, \u201cbut for speaking, you need to have courage. It must come from your own self. No one can help in courage own effort. In speaking, especially in presentation, it is difficult to improve in a group speaking. Because sometimes, someone will speak more and some will not speak at all. It's not about group. Also, to build confidence, we present alone, but only four or five people watch. It's not like we present many members and many people watch also, the courage is difficult to build\u201d (Participant 7).The next problem related to English language learning is the learners themselves. The barrier is often associated with learners' shyness and lack of confidence in using English, despite having learnt it since young. Also, they are afraid of being laughed at; hence they refrain from using English. One participant mentioned, \u201cHow do you learn English? What are the strategies that you use to learn English?Question: via texts emerged as the subthemes, only two participants mentioned it. Regardless of that, the researcher believed it is an exciting finding that should not be excluded from the report. One response mentioned, \u201cuse Thesaurus\u201d (Participant 4). Hence, the interviewer probed the answer:Third, the researcher asked what strategies the participants used in learning English. Though learning Interviewer: Could you explain how you use Thesaurus to improve your English?Participant 4: I find similar words. [long pause]Interviewer: Could you elaborate? Why do you use Thesaurus instead of Google?Participant 4: Google sometimes not good. I have it at home, the Thesaurus. Thesaurus has like synonyms and antonyms and got sample sentences.Reading novels, I like them. Sometimes I read Webtoons. My favorite\u201d (Participant 5). Webtoon is an online platform for comics, mainly from Korean authors, but the comics are officially translated to English. Though participant 4 preferred to use the hard copy version of Thesaurus, Participant 5 mentioned that she even read novels online because it's convenient.Another participant expressed, \u201cAlthough we [referring to him and his family] watch English movies, subtitles also English\u201d (Participant 6). Generally, participants learned English by watching and learning through the subtitles. Still, one response mentioned, \u201cI always watch movies and anime by reading the subtitles, and if I don't understand it, I would gladly Google translate the word\u201d (Participant 2).Audio\u2013visual materials like videos encourage learners to learn English. Many participants mentioned they watched videos on YouTube and other platforms in the English language. One interesting response said, \u201c, \u201cfrom games also we can know more words like vocabulary game\u201d (Participant 4). Another response also mentioned a similar answer, \u201cI use game word search also. I get to know more words\u201d (Participant 6). On another note, one response elaborated using video games instead of word games, \u201cand I used to expose to English at a young age since my nephew and uncle play video games so much, I started to play it and started to understand English a little bit\u201d (Participant 2).Another way of learning English, which various participants mentioned, was through games. One response saidvia materials, one participant specifically mentioned that learning English in class with a teacher's guidance helped him improve. He noted, \u201cOther than that, the learning process in class is also important to make sure our grammar, pronounce and literature good and nice. If somehow students out there didn't seem like to give a full attention on classes, maybe they won't be able to improve their English\u201d (Participant 2). Though only one participant mentioned this, it is worth noting as learning English with proper guidance helps.Though participants agreed that they learned How ready are you in mobile learning? What are the contents you require in a mobile application?Question: of course. Now no one wants to hold a heavy dictionary\u201d (Participant 6). Aside from the easiness to use a mobile app, Participant 2 mentioned that people are more attached to gadgets in the current days, which will be beneficial if an app for English is created. He specifically said, \u201cyes, because people nowadays are more attached to mobile phones, laptops, tablets, and PC more than they attached to books. So, it's really helpful for them to have a good app to improve their English\u201d (Participant 2). Additionally, Participant 1 personally mentioned the current pandemic situation, which makes learning through an app more beneficial through a mobile app. She stated that \u201cyes, of course. Because now during pandemic, students do nothing at home. So, for those who want to learn English, think English is difficult and want to improve their English, they can use the English apps\u201d (Participant 1).Finally, the researcher asked participants' readiness in using a mobile app to learn English. Participants are positive toward using the mobile app. All the participants are ready to use a mobile app as a supplementary lesson to improve their English language competency. Some of the reasons are the mobility feature, where learners can learn anywhere, which is easy. One response mentioned, \u201cit's important to put media like audios, videos, graphics, and animations\u201d (Participant 5). Another response also said that using audio\u2013visual materials, \u201cvideo can, but the important thing is not too long, like a minute okay maybe\u201d(Participant 5). Plus, one participant said, \u201cif we watch video, we can improve vocab\u201d (Participant 7). Aside from audio\u2013visual materials, many participants also mentioned games like quizzes and mini-games. Participants 3 and 7 noted that they are agreed on integrating quizzes in the app.Learning activities are central to improving learners' English language competency through the mobile app. One participant mentioned, \u201cfor example, if we play the tap app, understand? When you want to go to the next question or anything next, they'll ask you to tap this and that. Ah, that sounds like fun because it makes us feel like we want to know more, want to know more, like that\u201d (Participant 1). Additionally, Participant 2 also mentioned the fun aspects of a mobile app with mini-games, \u201c\u2026maybe mini-games, so that the app will not be very boring for everyone to use. A little fun would be okay\u201d (Participant 2).Participants 1 and 2 mentioned including mini-games to create a more engaging app. The suggestion from Participant 1 is unique as, she said, \u201cThis study aimed to explore the needs of STEM learners in creating the English language competency mobile module, adhering to HCI. The freedom to learn STEM subjects in different languages might cause difficulties for learners in the future Ye, . With reAdditionally, this study showed that many STEM learners mentioned being anxious and shy to speak English. Supporting the findings, a study that looked into the factors affecting English language learners' shyness reported that they had limited knowledge of the language, resulting in anxiety (Ahsan et al., via HCI could embed the social constructivism theory by Vygotsky (Human\u2013computer interaction prioritizes multiple interaction patterns and the openness of the design space. Participants in this study mentioned the use of audio\u2013visual materials. However, one participant also specified the teacher's involvement. Although the focus is on technology in HCI, there is a need to look into human interaction. Therefore, findings from this study clearly showed the importance of HCI to ensure that teachers' presence is also an aspect to be considered when developing an app. In enhancement, app development Vygotsky  to incluVygotsky . HoweverVygotsky . This prvia a mobile app, which can be considered when designing and developing feasible mobile applications.This study was carried out to explore the needs of STEM learners in creating an English language competency mobile module, which looked into specific samples with specific backgrounds. There are limitations to our study. First, there is a limitation in transferability. Also, the number of participants who volunteered was from one class in a public school; hence the sample might not represent the whole population of STEM learners. Despite the limitations, this study has strengths. First, this study is one of the earliest studies to look into the needs of STEM learners in second language learning from the HCI perspective. Second, this study provides evidence that learners have their preferences in learning via HCI.One major significance of this study contributes to the language policy aspect. Many non-native countries teach English as either a second or foreign language, especially in higher education, because English is an international language and learning English is necessary for education, especially for learners to further their education (Wallace and Lee, This study proposed exploring the needs of STEM learners in an English language competency mobile module. The results from the interview implied that STEM and English language rely on each other because language ability is an important aspect. This study also implied the importance of introducing vocabulary to second language learners first before teaching other language skills. This is because vocabulary is the foundation of language learning. Furthermore, findings from this study implied that further research on HCI to assist vocabulary learning is feasible, opening up more opportunities for future research on HCI.This study is significant for mobile app designers, English language teachers and course designers as the findings provide an overview of the STEM learners' needs in the English language. This study looks into learners' perspectives in mobile learning, which gives more effectiveness to HCI, rendering success to second language acquisition. One notable finding from this study, which is the mobile readiness of learners, provides insight into future mobile learning research. This study is only a preliminary study, and future work could look into the HCI aspect in designing and developing a mobile app to enhance STEM learners' English language competency.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent from the patients/ participants or patients/participants legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.KRMR, HH, and MMY conceived the study, participated in its design and coordination, and performed final analyses and co-drafted the manuscript. KRMR collected field data, entered study data, assisted in data analysis, and interpretation of study results. All authors read, revised, and approved the final manuscript.Universiti Kebangsaan Malaysia funded this research with Grant No. GG-2021-003.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "Self-regulation is important in enhancing students\u2019 academic performance, yet evidence for the systematic and valid instruments to measure self-regulated learning strategies of college students in an English as a foreign language context is far from robust. This study was situated to develop an evaluation tool to examine the status quo of self-regulated learning strategies employed by college English learners and the associations between the use of these strategies and their academic achievement. A large-scale survey was conducted at a university in Macau to provide evidence of the construct validity of responses to the questionnaire on self-regulated learning strategies. Conceptualized in social cognitive theory, the questionnaire comprised environmental, behavioral and personal self-regulated learning strategies with 48 items weaving into 10 dimensions. Strong evidence for reliability and validity was found. Findings also revealed that students who intrinsically valued and used more self-regulated learning strategies achieved higher academic performance. Students in advanced-level English course reported significantly more frequent use of self-regulated learning strategies than students in medium-level and mixed-level English courses. Our results draw attention to the pedagogical orientation for teachers of English as a foreign/second language in helping students become adaptive learners with self-regulative process. Since its inception, self-regulation has been accorded great importance as a 21st-century skill . SchoolsPrevious studies on SRL strategies have laid a promising theoretical foundation, but the validity evidence for a systematic instrument to measure college students\u2019 SRL strategies in English as a foreign language (EFL) contexts is far from robust . It is padaptive learners with a range of self-regulative processes through which they set goals, manage resources, self-monitor, and seek feedback; (b) positive learners who sustain learning interest and show confidence in achieving learning objectives; and (c) proactive learners who know how to select the best strategies to suit their abilities, based on their self-motivational belief in their strengths and weakness , learners clarify and share the goals and standards to attain in a certain task. This phase involves students\u2019 perception of the task\u2019s affordances and constraints and their motivation arising from beliefs about learning such as self-efficacy and outcomes expectancy (performance), learners engage with the task and monitor their learning, usually deploying planned strategies to compare their progress against standards set in the forethought phase and discover causes of learning events. In the third phase (self-reflection), learners evaluate their work and generate applicable revisions or adjustments thereto. This includes reflecting on feedback and mentally storing ideas and concepts to use in the task. To conclude, self-regulated learners make deliberate and goal-directed efforts to adjust, adapt, or abandon their learning strategies and identify, retrieve, and seek new information for future learning , evaluating learning performance (self-judgment), and responding to performance outcomes (self-reaction). In a meta-analysis of SRL by The most comprehensive set of self-regulatory strategies  has been widely discussed in the sphere of language learning . PublishOxford and colleagues reported a broader range of SRL strategies, with six sub-scales: memory strategies, cognitive strategies, compensation strategies, metacognitive strategies, affective strategies, and social strategies . In the via empirical investigations in the Macau SAR. Such investigations can provide insights to educators in other places similar to Macau.Although a plethora of research has discussed the link between SRL strategies and learning achievements, several issues remain. Concerns include theoretically important but empirically uncertain questions regarding the use of SRL strategies among college students in the Macau context, the variables affecting their English language proficiency, and the lack of a valid and reliable instrument to measure their SRL strategies for EFL education. An investigation into these factors would be theoretically intriguing and is urgent for the context of the present study, the Macau Special Administrative Region of China. The fundamental law of Macau education system has placed great emphasis on more quality-concerned English classes and launched the appeal of \u201cnurturing students\u2019 attitude and ability of life-long learning\u201d . Over thInvestigating the challenges and factors that may affect foreign language learning is pivotal for informing theory, practice, and policy. In a highly dynamic and constantly changing educational landscape, researchers and teachers more than ever need to dissect the field\u2019s challenges and move forward to best serve the shifting needs of language learners. Informed by research from recent decades, de facto function , opportunities to interact with these languages in everyday settings are prodigiously few with large class size  were enrolled in a required Level One English course, 42% (n = 250) in a Level Two course, and 10% (n = 62) in two mixed-level classes for students admitted in or before the 2018/2019 academic year. These students spend 2 h each week in class and approximately two additional hours each week doing homework for the English class. Approximately 43% of the participants were male (n = 255) and 57% were female (n = 343). Participants\u2019 ages ranged from 16 to 26  and their years of English learning ranged from 0 to 23 years . The participants were from diverse academic faculties, including School of Business, Faculty of Law, Faculty of Chinese Medicine, Faculty of Hospitality and Tourism Management, University International College, and Faculty of Humanities and Arts. The context of leveled and mixed ability groups provided accountable data for further multi-trait comparisons.Volunteer participants were recruited from Macau University of Science and Technology during the first semester of the 2020/2021 academic year. The study was approved by the Research Ethics Committee and carried out in accordance with the institutional requirements. The researchers obtained informed consent from all students before conducting the survey. Meticulous attention was paid to research consent, benefits, privacy, and confidentiality, and participants were informed of their right to withdraw from the study at any time. The sample comprised 598 undergraduate students from 22 classes. Approximately 48% (Gaokao or the Joint Admission Exam (both are further elaborated in the following section). As the research was carried out in the autumn term, students with the highest proficiency level were enrolled in Level Two and students at the medium level were in Level One. The two mixed-level classes primarily contained students who had failed either Level One or Level Two in a previous academic term. Lecture periods typically comprised a weekly 3-h session for Level One and Level Two and two 2-h sessions for the mixed-level course.The overwhelming majority of Level One/Two participants were in their first year, while the students in the mixed-level classes were juniors or seniors. All students were required to earn required general English education credits in their first academic year, but the university sorted them into different course levels based on their English language test in the This research progressed through three stages. At the first stage, the conceptual stage was oriented toward questionnaire item generation by exploring the existing literature and consulting with several well-versed practitioners in the field. A pilot study was then conducted in the second stage to determine the framework of ESRLQ. A subsequent interview with 10 first-year students from the 2019/2020 academic year was used to confirm that the items in ESRLQ were appropriate for this population. The third stage involved scrutiny of the psychometric properties of the updated ESRLQ using confirmatory factor analysis.A feasible and reliable questionnaire from In line with The research team received participants\u2019 permission to access their official English final grades at the end of the semester. Participants\u2019 academic achievement was evaluated based on their final reading test score, taken from university records. The grade scale ranges from a minimum of 0 to a maximum of 100, with higher scores indicating greater English language proficiency. For each final, members of the panel of expert or experienced tertiary educators are invited to collaborate in designing the test. They hold meetings to scrutinize the course objectives and make sure the test martial must cover all relevant parts of the course it aims to measure, so as to maintain the norms and the content validity of the exam. The average score of the participants on this exam was 62.56 with a standard deviation of 18.97.Gaokao, a standardized test administered annually in mainland China, is commonly known as the national university entrance exam and also viewed as China\u2019s version of the American SAT and British A-level exams. One of the mandatory Gaokao exams is the English language test. The English language test consists of three sections  and the full mark of English language test accounts for 150 in most places while only a few provinces have 120 in total score. Gaokao English results are important because students with a higher score are assumed to be more proficient in the language, which increases the possibility of being enrolled to a top-tier university. Notwithstanding the fact that some top test-takers in Gaokao English exam may be questioned about their practical communication competence with native speakers, universities still accept the score as a basis for direct entry and it also witnesses an uptrend in western institutions to take Gaokao result as a measure of academic competence .mpetence . The valts e.g.,  and cons average . Gaokao JAE) represents a local student\u2019s language proficiency as it is the admission examination jointly organized by four higher education institutions in Macau . Only Chinese, Portuguese, English, and Mathematics are on the JAE; other subjects are tested by institutions individually. The JAE English language test (JAE-E), per the announced syllabus, corresponds to the Common European Framework of Reference .The 120-min test is composed of Language Use (40 marks), Reading Comprehension (30 marks), and Essay Writing (30 marks); exam questions are set at a variety of levels, including Elementary (CEFR A2), Pre-Intermediate (CEFR B1), Intermediate (CEFR B2), and Upper-Intermediate/Advanced (CEFR C1). The final mark received by the local student in Macau is generally a weighted sum of their subject marks . Every year, around ten academic members from the four institutions constitute a committee to jointly organize and administer the exam. To understand the construct and content validation of Gaokao and JAE, all the raw scores were transformed into z-scores and then back-transformed into standardized scores with the same mean (62.56) and standard deviation (18.97) with the final English language Examination. This is reducing the chance of the violation of homogeneity of variance in statistical analysis. The transformed scores keep the rankings of the raw scores.As the range of scores differ between the The structural aspect of the validity of participants\u2019 responses to the survey was checked with Confirmatory factor analysis (CFA). Instead of using Gaokao/JAE English test score for comprehensive English language proficiency and the final English exam score measured at the end of the semester for English language reading competence. Multivariate analysis of covariance (MANCOVA) was employed to compare the SRL strategy use as well as the comprehensive English language proficiency and English language reading competence between the three cohorts: Level 1, Level 2, and Mixed Levels. The number of years studying English was used as a covariant. Effect size  was reported as small (0.01), medium (0.06), or large (0.14) according to Pearson correlation coefficients were used to check the external aspects of the construct validity by correlating the use of SRL strategies measured by ESRLQ with student performance on two English examinations: r = 0.47, p < 0.001, as well as student\u2019s English language reading competence, r = 0.46, p < 0.001, both with a large effect size . However, participants reported the least use of seeking opportunities to practice the English language although the use of this strategy was strongly associated with the comprehensive English language proficiency (r = 0.40) and English language reading competence (r = 0.41).Descriptive statistics of the use of SRL strategies by students in the three cohorts are in r = 0.30, p < 0.01. As a result, the assumption to use MANCOVA to compare the SRL strategy use between the students from the three cohorts: Level 1, Level 2, and Mixed Levels was met. Statistically significant differences were found in the linear combination of all three dependent variables, Wilks\u2019 lambda = 0.50, F = 76.36, p < 0.001, partial \u03b72 = 0.29 (large effect). Tests of between-subjects effects suggested statistically significant differences in the use of SRL strategies, F = 47.62, p < 0.001, partial \u03b72 = 0.15 (large effect); comprehensive English language proficiency, F = 258.92, p < 0.001, partial \u03b72 = 0.48 (large effect); and English language reading competence, F = 64.44, p < 0.001, partial \u03b72 = 0.19 (large effect). Post hoc multiple comparisons noted that students in Level Two (advanced level) scored more than their peers in Level One (medium level) and that students in Level One scored more than their counterparts in Mixed Level (those who failed in Level One or Level Two) in all three dependent variables, namely, use of SRL strategies, comprehensive English language proficiency, and English language reading competence.The linear relationship between the number of years studying English and the English language proficiency score was statistically significant, SRL theory has been promoted to examine and foster students\u2019 functioning in academic contexts . The curFuture researchers may adopt this instrument to measure Chinese college students\u2019 use of SRL strategies, especially in the region of Macau (and maybe Hong Kong) where student characteristics differ from those in mainland China. Students in Macau and Hong Kong are similar to students in mainland China in that they are all Chinese in ethnicity but different from students in mainland China in that they inherit quite a lot western culture due to the history. Another difference is that English is used much more often in both school settings and social context in Macau and Hong Kong.Also noteworthy was that in our study, advanced-level students reported significantly more frequent use of self-regulated learning strategies and scored higher than their peers in comprehensive English language proficiency test and English language reading competence test. The positive relationship between the use of SRL strategies and academic achievement reported in this study echoed previous research with learning strategies in the field of teaching/learning English as a foreign/second language e.g., .Empirical evidence emerged in our research also supports the social cognitive theory  and prevThe findings of our study provided evidence for the reliability and validity of student responses to the survey to measure the use of SRL strategies. First, the internal structure of the instrument to measure the use of SRL strategies remained consistent with the previous version from which it was adapted . The extDescriptive statistics of the use of SRL strategies suggest that college students in Macau use some SRL strategies but not very often. Strategies under the category of Seeking Opportunities  were least used. One possible reason is that these strategies are more related to students\u2019 intrinsic motivation rather than teachers\u2019 requirement in English reading courses. While societal norms and cultural values in Macau may have an impact on students\u2019 learning intention and behavior\u2014in absence of a positive sense of wellbeing, students tend to follow teachers\u2019 request and are not often trained to take initiatives for their learning . This imEnglish language instructors may need to adopt SRL strategy development approach to help students gain more SRL strategies. A recent meta-analysis of 22 primary studies has provided a strong and positive link between the use of SRL strategies and language learning outcomes . In thisThis study also has practical implications for teachers of ESL/EFL. The findings of this study echoed previous empirical studies and provided evidence to support the strong and positive associations between the use of SRL strategies and English language proficiency. Teachers of English are encouraged to participate in professional development workshops and to learn how to explicitly and systematically incorporate the use of SRL strategies in their classroom instruction so that EFL/ESL learners can benefit more from the English language course. With the help of their teachers, ESL/EFL learners may become adaptive learners with self-regulative process who know how to set goals, manage resources, and monitor their progress, positive learners who may sustain interest and maintain confidence, and proactive learners who know how to select the most appropriate strategies for themselves , 2008. TGaokao like other Chinese students do. The JAE test was specially tailored for them. Therefore, test equivalence between Gaokao and JAE is another direction for psychometricians in the future. Finally, readers should be cautious when interpreting the results and generalizing the findings to their own population .Future researchers are encouraged to have a closer examination of the differences between deep and surface cognitive strategies, which was not examined within the current study. Another limitation of this study lies in the small target population. Although the students in Macau were rarely studied in the past, they are a special Chinese group because they do not have to take The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Macau University of Science and Technology. The participants provided their written informed consent to participate in this study.XD: data collection and manuscript writing. CW: conceptualization, data analysis, and reviewing and editing. JX: reviewing and editing. All authors contributed to the article and approved the submitted version."}
+{"text": "The present study aimed at investigating the motivational factors affecting Iranian learners\u2019 learning of English as a Foreign Language across differing levels of language proficiency. To this end, 110 males and 70 females with an age range of 18\u201331 took part in the study and a mixed-methods approach was adopted. First, the researchers administered Oxford Placement Test (OPT) to determine the proficiency level of the participants and placed them into three levels of language proficiency. Elementary, intermediate, and advanced. Next, as for the quantitative part of the study, the Attitude/Motivation Test Battery (AMBT) questionnaire was distributed among all the participants to delve into the motivational factors. Finally, a semi-structured interview was conducted to qualitatively probe into EFL learners\u2019 motivational factors affecting their learning of English. It was found that the motivational factors which encouraged EFL learners resulted from either intrinsic or extrinsic tendencies. The recognized intrinsically-based motivational factors were personal enjoyment, social prestige, and being challenged. The extrinsically-oriented factors as the source of motivation to learn English were getting a job, pursuing education, earning more money, traveling to foreign countries, pursuing academic careers, making efficient use of technology, and effectively interacting with native speakers. It was concluded that beginner and intermediate English language learners tended to learn English more based on extrinsic factors whereas advanced learners were found to be more intrinsically motivated. Motivation plays a pivotal role in the psychological and pedagogical settings. However, attempts to theorize motivational factors through a single mechanism have failed and are mainly done away with . NeverthMotivational factors affect the choice of language learning strategies such as memory, socio-affective, and cognitive ones . Zheng eAlthough, motivation and some of its aspects has been investigated abundantly in the literature e.g., , the motFrom the early beginning of the 20th century, motivation has been abundantly dealt with in the literature. Despite developing many theories and research, motivation is still a controversial topic and remains a bone of contention among many researchers and practitioners within the field of applied linguistics. Many scholars used the most known theories and models of motivation as a starting point for their work within the realm of motivation. Armstrong (2007) summarized those theories in his book about employee reward management. He argued that Taylor\u2019s theory of motivation to work is related to compensations and penalties which are directly connected to performance. A less instrumental approach is Maslow\u2019s concept of the hierarchy of needs which describes motivation emanating from people\u2019s unsatisfied needs. The distinction between extrinsic and intrinsic motivators was the study of Herzberg. Those \u201cold\u201d theories are important but not perfect .Emphasizing what motivates people, requirements that are the same for everyone. Despite assuming a similar set of needs for all people, they define the needs and requirements of everyone differently. Maslow\u2019s theory about human needs is the most well-known in the field of motivation. He states that human behavior is driven by the existence of unsatisfied needs .A dynamic character characterizes the process theories, not static as content theories. What motivates people is the main concern, not how motivation occurs. Process theories explain the way of behavior and attitudes of people which are aimed at certain options. Process theories focus on individual\u2019s mental processes in specifying the motivational level .Self-determination theory (SDT) is one of the most all-embracing and inclusive motivational theories, the primary tenets of SDT are explained and delineated by Going through the related literature, it seems that there exists a paucity of research dealing with motivational factors that affect EFL learners\u2019 learning of English in the Iranian context at differing levels of language proficiency. More specifically, most of the previously-done studies have adopted either a quantitative approach or in few instances a qualitative approach separately without adopting a combination of both to delve into the motivational factors. Moreover, not only did the study intend to adopt a mixed-methods design to augment the overall validity and reliability of research findings, but also it went even further to see whether any discernible pattern of motivational factors existed for EFL learners across differing levels of language proficiency. Therefore, the researchers formulated the following research questions to meet the above-stated objective of the research:(1)What motivational factors influence Iranian EFL learners\u2019 English language learning?(2)What motivational factors influence the Iranian EFL learners\u2019 English language learning across different English proficiency levels ?(3)Are there any common motivational themes among Iranian learners at different levels of language proficiency?(4)Is there any significant difference among Iranian elementary, intermediate, and advanced EFL learners in terms of the motivational factors which influence their English learning?Two-hundred Iranian EFL learners studying English in Gooyesh, Rohroshd, and Safir Language Institutes in Qom, Iran took part in the study. Due to the special conditions imposed by the COVID-19 pandemic, from among this number, 180 EFL learners  from different language proficiency levels  could complete the entire course of the study. Their age ranged from 18 to 31. The demographic information of the participants is presented in r = 0.86), which was fairly satisfactory. Furthermore, to ensure its validity, it was reviewed by two language experts and their comments were applied in the follow-up version of the main study. The OPT consisted of three parts: Vocabulary Test, Reading Comprehension, and Grammatical Structures. This test included 40 vocabulary tests, 40 grammar tests, and 20 reading comprehension tests.To determine the participants\u2019 language proficiency level, the researchers utilized Oxford Placement Test (OPT) at the beginning of the study. Also, using the KR21 formula, the internal consistency of the test was calculated and reported : One full professor and two associate professors. They were kindly asked to go through the related questions and determine whether they were appropriate based on the participants\u2019 proficiency level and the purpose of the study. Accordingly, a pilot study was conducted before the main study to probe into the potential drawbacks of the questions.The questionnaire was adopted from Gardner\u2019s Attitudes/Motivation Test Battery (AMTB). This questionnaire is composed of 10 multiple-choice items on a 5-point Likert scale. Furthermore, to make sure about the reliability of this research instrument, the researchers conducted a pilot study with 30 participants before starting the main phase of the study and the obtained coefficient was found to be 0.79 which is considered an acceptable and satisfactory level of internal consistency.The following steps were taken to achieve the already-stated objectives of the study. First, the researchers administered OPT to determine the proficiency level of 200 participants and placed them into three levels of language proficiency: Elementary, intermediate, and advanced. Next, as for the quantitative part of the study, the researchers distributed the AMBT questionnaire among all the participants to delve into the motivational factors that could have affected their English learning. Finally, a semi-structured interview was conducted to qualitatively probe into EFL learners\u2019 motivational factors affecting their learning of English.Having recorded all EFL learners\u2019 responses to the semi-structured interview questions, the researchers went through the recorded data as thoroughly and meticulously as possible and transcribed the recordings to determine the most typical themes within the transcriptions. The thematic analysis was done based on To answer the research questions for the quantitative phase, the researchers inserted the results of the OPT into the SPSS software and employed both descriptive and inferential statistics. As for the qualitative data analysis, the obtained information from the semi-structured interviews was transcribed, coded, and analyzed. A systematic guideline for data coding was adopted from To investigate whether the interview items enjoyed an acceptable level of reliability, the researchers ran inter-coder reliability. The results of this analysis are summarized in As demonstrated in \u201cThe extent to which an instrument looks as if it measures what it is intended to measure\u201d . Direct Prior to the administration of the questionnaire for the main study, the researchers piloted it with 30 EFL learners. The obtained data were analyzed using Cronbach\u2019s alpha method to find out the reliability coefficient.As shown in The participants\u2019 answers to seven interview questions about motivational factors were categorized into nine distinct themes which are presented in \u201cLearning English skills brings about a sense of satisfaction and happiness.\u201d Concerning being challenged, one of the participants stated: \u201cI am really motivated to engage in challenging tasks with an optimal level of difficulty.\u201d Furthermore, concerning the communication theme, another participant answered that: \u201cI like to get in touch with some native individuals from English-speaking countries through emails or other social networks.\u201d Regarding pursuing academic purposes, one of the participants asserted that: \u201cI\u2019d like to attain the highest score possible in IELTS and TOEFL tests and be awarded such important certificates.\u201d Within the employment theme, another participant responded: \u201cEnglish helps me to develop and improve my business by being able to understand English sources, texts, and materials.\u201dAccording to The second research question sought to determine the motivational factors which might influence the elementary EFL learners\u2019 tendency to learn English as a foreign language. The elementary level participants\u2019 responses to the interview questions were analyzed and the results of thematic analysis are presented below.\u201cLearning English brings about a kind of high social class and prestige for me in front of my friends and relatives.\u201d Another participant who learned English for enjoyment and entertainment stated that: \u201cI am very interested in learning and knowing about a language other than Persian.\u201dAs shown in Learning English helps me to work with my PC much more easily and efficiently.\u201d Concerning finding job opportunities, another participant stated that: \u201cI\u2019m going to be able to teach English one day and make money with that.\u201d Yet another elementary level EFL learner responded that: \u201cI\u2019d like to be able to speak fluently with an English native speaker.\u201dFurthermore, the elementary level participants reported three extrinsic Factors to which their tendency toward learning a foreign language could be attributed. They pointed to the efficient use of new technologies, facilitation of employment, and making effective communications as the extrinsically-oriented factors that encouraged them to learn English. For example, one of the participants reported that: \u201cMost non-proficient and elementary learners who were at the initial stages of EFL learning wanted to learn English based on extrinsic factors especially their needs to find a decent job, to earn money and to interact with individuals who live in an English-speaking country. Moreover, from among these elementary EFL learners, a few considered intrinsic factors as their main source of motivation and primary driving force to learn English.The second research question also sought to specify the motivational factors which influenced the intermediate EFL learners\u2019 motivation to learn English as a foreign language. The results of a thematic analysis of these learners\u2019 responses to the interview items are illustrated in \u201cI really like the social class and prestige I gain through learning English and I feel more confident in the community I live in.\u201d The other participant who learned English to meet personal interests and find enjoyment said that: \u201cWhen I speak English fluently with a tourist, it makes me proud of myself.\u201d Concerning the extrinsic factors, the intermediate EFL learners reported three extrinsic factors which affected their motivation to learn English. To the intermediates, educational needs, employment opportunities, and making communication were the major driving forces to engage in learning English. For example, one of the participants responded that: \u201cMastery of English vocabulary and grammar would greatly contribute to my likely success in the university entrance exam.\u201d With regard to finding a decent job, another respondent stated that: \u201cI\u2019m going to be able to teach English in a private English language institute.\u201d With regard to communicating with English native speakers, a participant said that: \u201cI\u2019d like to be able to communicate well with an English native speaker.\u201dThe information in Most intermediate English learners who were considered to possess a relatively good command of English language skills and proficiency were more extrinsically-motivated. Finding a decent job, pursuing academic careers, and communicating well with native speakers were among such motivational factors. Moreover a few intermediate language learners regarded intrinsic factors as the primary source of motivation and tendency to learn English.The last part of the second research question intended to explore the motivational factors which underpinned the advanced EFL learners\u2019 disposition to pursue learning of English at higher levels of language proficiency. This group of the participant\u2019s responses to the interview questions were analyzed and the results of thematic analysis are demonstrated in \u201cLearning different aspects of a foreign language is a deeply enjoyable and pleasant experience for me.\u201d However, this group of participants reported three extrinsically-oriented factors to which their desire to learn English could be attributable. They introduced and recognized the facilitation of fulfilling academic purposes, provision of good employments; and migration opportunities as the extrinsically-oriented motivations which might direct and encourage them to improve their English proficiency level as much as possible. For example, one of the respondents said that: \u201cI\u2019d like to receive as many English learning and teaching certificates as possible.\u201d Another one contended that: \u201cI\u2019d like to be admitted to a reputable university in Canada and move to this foreign country.\u201d Accordingly, it can be said that most advanced EFL learners reported such extrinsic motivating factors as pursuing academic careers, finding job opportunities, and migrating to an English-speaking country as the secondary driving forces to learn English.As demonstrated in The third research question aimed at examining the motivational factors underlying the participants\u2019 tendency to learn English which was common among the three groups of EFL learners in terms of their proficiency level. In all three groups of EFL learners, i.e., elementary, intermediate, and advanced, both intrinsically and extrinsically-oriented factors were found to play an effective role in fostering their tendencies toward English learning. More specifically, the analysis of their responses to the interview questions indicated some similarities and differences. Concerning the intrinsic factors, the \u201cpersonal enjoyment\u201d was reported by all three groups of the participants as one of main rationale behind English learning. In the same vein, social prestige was also found to be a common factor that might provide ground and motivation to learn English among elementary and intermediate EFL learners. On the other hand, concerning the extrinsically-oriented factors, finding job opportunities was found to be a common factor which the participants across all three groups of language proficiency expressed as their driving force to learn English. More precisely, when comparing the elementary and intermediate EFL learners, not only were finding job opportunities reported by both beginner and intermediate language learners as one main driving force to pursue learning English, commutating well with native speakers was also another motivating factor that were alluded to by both groups of language learners. In other words, elementary and intermediate participants had unanimously agreed upon these two extrinsic factors. To sum up, it was found that personal enjoyment and finding decent job opportunities were the two motivational factors which all the participants, regardless of their English proficiency level, declared as the major driving forces to pursue their English learning.To further investigate the motivational factors and their differences in terms of participants\u2019 English proficiency level, the obtained data from the motivational factor questionnaire was analyzed and used to answer the last research question of the present study quantitatively.The fourth research question of this study intended to investigate quantitatively whether the participants in three proficiency groups  were reported to be motivated by different factors in learning English. To answer this research question, a one-way ANOVA analysis was conducted to draw a comparison among the three groups. The descriptive statistics of the participants\u2019 self-perceived motivational factors are summarized in M = 4.49, SD = 0.98) and the lowest mean score  to the advanced level participants. To ensure that the differences are statistically significant the data in As indicated in p < 0.05 level in driving motivational factors to English learning for the three groups: F  = 38.21, p = 0.001. In addition to reaching statistical significance, the actual difference in mean scores between the groups was also large. The effect size, calculated using eta squared, was 0.75. As displayed in post hoc comparisons using the Scheffe test indicated that the mean score for Group 1  was not significantly different from Group 2 . While, it was found that Group 3  did differ significantly from both Groups 1 and 2. In other words, the findings indicated that the EFL elementary English learners were not significantly different from the intermediate English learners in terms of the driving forces that pushed them forward to learn English.There was a statistically significant difference at the This study was set to delve into the motivational factors that affected Iranian EFL leaners across differing levels of language proficiency. To pursue this line of inquiry, a concurrent triangulation mixed-methods design was adopted in which both quantitative and qualitative data were concurrently collected to provide thorough responses to the research questions.The first obtained results from the qualitative parts of the study suggested that for all the participants the motivational factors which might encourage and direct the EFL learners to learn English can stem from either intrinsic or extrinsic tendencies and orientations. Advanced learners were reported to resort to intrinsic factors as the driving forces for learning English. They declared some factors such as personal enjoyment and being challenged as the reasons for which they would like to pursue their English learning process. Beginners and intermediate language learners expressed the extrinsically-oriented factors as their source of motivation toward English learning and pointed to reasons such as getting a job, pursuing education, earning more money, traveling to foreign countries, fulfilling academic purposes, making efficient use of technology, as well as making effective interactions with native speakers as the motivational factors which might encourage them to learn English. However, majority of elementary and intermediate English language learners were more extrinsically-motivated to learn English and their underlying reasons to pursue English were primarily extrinsically-oriented.The second major qualitative finding of this study suggested that many novice English learners tended to learn English based on extrinsic factors, especially employment and communication requirements. Moreover, the obtained results from qualitative analysis indicated that the participants, regardless of their proficiency level, unanimously considered personal enjoyment (as an intrinsic factor); and getting employment opportunities (as an extrinsic one) as the two significant factors which underlay their motivation to learn English.The qualitative finding in this section seems to be partly in agreement with that of To further investigate the qualitatively identified differences between the three groups of participants  in terms of the driving motivational forces to learn English, a quantitative analysis was also conducted using ANOVA statistics. It was found that despite the reported distinctions in the qualitative results, in the quantitative analysis, there was only a significant difference between the advanced EFL learners and their elementary and intermediate counterparts with regard to the motivational factors affecting their English learning. Such motivational differences across varying levels of language proficiency can be attributed to the purposes, perspectives, environment, and family class. More precisely, the educational and schooling system in Iran necessitates that both teachers and learners to resort and being largely accustomed to largely to the traditional procedures of language teaching, testing, and learning in which the main focus is on the prescribed and passive teaching and learning materials. The students in this learning atmosphere are not expected to show enough motivation, self-confidence, and interest to engage in innovative learning activities in the classroom and obtain desired goals and objectives. A great number of families even teachers and students are not satisfied with the achievements of English learning. The low quality and ineffectiveness of the schooling system seem to be among the most crucial factors that exacerbates the this dissatisfaction.On the other hand, the commonality of employment and personal enjoyment as motivational factors among Iranian learner\u2019s English learning could be considered an expected result which might not come as a surprise to some extent. Iran\u2019s increasingly tight job market, as a result of poor economic policies, as well as global sanctions turn employment into a big challenge for young people. Therefore, one of the most prevalent driving forces among the Iranians, specially the young educated ones to pursue an academic career is to get a decent job and be given a good and proper job opportunity. Learning a foreign language like English is no exception.In this study, a mixed-methods design with a concurrent approach was adopted to explore and compare the categories of motivational factors which foreground the EFL learners\u2019 tendencies toward learning English as a foreign language. The obtained data from a motivation questionnaire and interview questions were analyzed using some inferential statistics and thematic analysis. It was found that the motivational factors which direct the EFL learners to English learning can result from either intrinsic or extrinsic tendencies. The recognized intrinsically-based motivational factors were personal enjoyment and being challenged which were considered as the reasons for which they would like to pursue their English learning process. Beginner and intermediate language learners expressed the extrinsically-oriented factors as their source of motivation toward English learning proposed reasons such as getting a job, pursuing education, earning more money, traveling to foreign countries, pursuing academic career, making efficient use of technology, and effectively interacting with native speakers as the motivational factors which might motivate them to learn English. Furthermore, it was concluded that a large number of novice English learners tend to learn English based on extrinsic factors, especially employment and communication requirements. Intermediate EFL learners\u2019 tendency to learn English could be attributed to being successful and getting the desired achievement in education, providing a lot of employment opportunities, and making effective interactions with native speakers. Furthermore, most of the advanced learners considered academic purposes, employment, and migration as the main reasons which might underpin their tendency to maintain learning English.Moreover, educators should be more cautious in choosing the right instructional materials and pedagogical tools. To decrease demotivation, teachers are recommended to pay due attention to the proper selection and adoption of teaching methodology and their manner that can boost learners\u2019 motivation. Furthermore, teachers need to integrate all language skills, instead of vocabulary and grammatical rules at the expense of oral skills. To motivate language learners, teachers need to take into account the contextual elements and learners\u2019 personal experiences as well.In the same vein, materials developers are highly recommended to revise instructional materials to increase students\u2019 motivation. Learners\u2019 negative perceptions about and attitudes toward the second language could be changed by promoting the content of textbooks. English textbooks might fail to attract the learners\u2019 interests and decrease their motivation by putting too much emphasis on grammar and vocabulary. Instead, textbooks should provide opportunities for classroom interaction and enable educators to improve learners\u2019 overall oral skills. Accordingly, materials developers need to develop intriguing tasks and topics for the texts in their textbooks to promote learners\u2019 motivation. Regarding students\u2019 negative beliefs and attitudes toward the English speaking community as a demotivating factor in learning English, learners\u2019 motivation can be readily boosted through incorporating cultural information in students\u2019 English textbooks. For instance, cultural tasks can be appropriately included in the language lessons to improve the quality of instructional materials. Educators and instructors need to play their pivotal parts as motivators to students who are struggling to learn a second or foreign language.In the Iranian educational system, the English curriculum often focuses on preparing L2 learners to pass their exams. English instructors can pay more attention to the communicative facets of the English language in their instructional practices. In this regard, English teachers can maintain and enhance learners\u2019 L2 motivation through various activities in such pedagogic books. This study might provide practitioners and educators with some useful insights into English planning and policy, and it would also offer suggestions for English syllabus designers and material developers in Iran that are congruent with L2 learners\u2019 motivational orientations. Finally, this study might help enhance the organization and presentation of current English-language academic sources in various pedagogic settings.Nevertheless, some limitations should be taken into account, and caution should be exercised in interpreting the findings of the qualitative part of study because of the following reasons. First, generalizability of the findings could be undermined due to the included sample in the qualitative part. Second, because of the difficulty of unraveling and coming up with the most typical motivational themes across varying language proficiency levels and the relative subjectivity inherent in such thematic analysis, replication research is also required to corroborate the results of the present study. Finally, EFL learners might have suffered from a self-flattery syndrome and overrated themselves to project an ideal self-image of themselves when answering the items of the motivation questionnaire.The effect of adopting motivational strategies on the EFL learners\u2019 attitudes toward foreign language learning can be investigated. Second, the effectiveness of intrinsic versus extrinsic motivation strategies on language learning achievement can be explored among EFL students at tertiary level. Third, this line of inquiry can be pursued by investigating the correlation between extrinsic and extrinsic motivation with such variables as self-efficacy and anxiety. Finally, motivation is just one of the several elements that could affect the learning process and outcomes. Therefore, instructors need to closely attend to other pertinent factors such as students\u2019 cognitive style, personality factors and language aptitude to further engage them in the learning process. The motivational factors discussed earlier need to be complemented by further empirical research as well.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent from the participants\u2019 legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.RB was the corresponding author and contributed more than AF. RB as the corresponding author was the main contributor of this research paper. Both authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "The present study examined English use anxiety, motivation, self-efficacy, use of English, and their predictive effects on top university students\u2019 English achievements. Two hundred and twenty-three students of the Top-Notch Students of Basic Disciplines Training Program in a top Chinese university answered a battery of questionnaires, which consisted of the 8-item English Use Anxiety Questionnaire, the 5-item Motivational Self-Talk Questionnaire, the 3-item Self-Efficacy Questionnaire, the 19-item Language Learning Orientations Questionnaire, and a Background Information Questionnaire. Analyses of the data revealed the following major findings: (1) the participants had a low level of extrinsic motivation-introjected regulation, a low-to-medium level of English use anxiety, extrinsic motivation-external regulation, intrinsic motivation-knowledge, and a medium-to-high level of motivational self-talk, self-efficacy, extrinsic motivation-identified regulation, intrinsic motivation-accomplishment, and intrinsic motivation-stimulation, (2) use of English anxiety (UAE) and language learning orientation were generally significantly negatively correlated with each other, and significantly correlated with other measured variables, (3) UAE and intrinsic motivation-knowledge significantly predicted the participants English achievements, measured both by standardized test scores and self-rated overall English proficiency, and (4) use of English and self-efficacy mediated the effects of English use anxiety and language learning orientations on the participants\u2019 English achievements. These findings further pinpoint the importance of anxiety and motivation in second/foreign language learning. Many studies have proven that besides being influenced by teachers and teaching materials, second language (L2) learners\u2019 achievement can be affected by their own individual factors such as gender, age, anxiety, proficiency in the target language, self-confidence, motivation, and self-efficacy . It is gAs reviewed in more detail below, contact with the target language, foreign language anxiety, language learning motivation, and self-efficacy are all important factors in L2 learning and acquisition. However, they have rarely been examined in relation to L2 learning outcomes simultaneously in the same context. Moreover, most research in second language acquisition focuses on ordinary students, with a few studies targeting good/successful learners, little research can be found on top students of top universities. In order to prepare leading scholars in basic disciplines, China\u2019s Ministry of Education has been investing in top undergraduate students in top universities by providing them with (much) more learning and research resources during the recent decade. Nevertheless, little research has been done on these students to examine how they learn a foreign language. This is the aim of the current study, which sought to examine the profiles of English use anxiety, motivation, and self-efficacy of top students in Chinese top universities and how they affected the students\u2019 learning of English, the primary foreign language in China, hoping to shed light on general English teaching and learning in higher education in China and elsewhere of the world.Studies about anxiety of foreign language learners can date back to the 1970s, but it was not identified as one separate anxiety type until mid-1980s , when scForeign language anxiety can affect the second language learning process in various ways and may interfere all three stages of language input, processing, and output, being especially problematic during the two latter stages . HorwitzTo quantify L2 learners\u2019 anxiety, Even though positive psychology is catching researchers\u2019 increasing attention, anxiety is still an important factor affecting L2 learning and a topic of research in second language acquisition. And it is interesting to know whether and to what degree top students in Chinese top universities feel anxious in learning and using English.The relationship between learning attitudes and second language achievement has always been what researchers are interested about . GardnerIt is not easy to relate the complicated construct of motivation to actual behaviors and build up a certain single model, let alone the specific motivation of learning a second language . AccordiDespite the plethora of research on L2 motivation, more continuous research is needed on students with various backgrounds, since motivation is dynamic and is continuously shaped by the learning environment .The concept of self-efficacy was introduced by At the same time, self-efficacy can also influence L2 learner\u2019s learning experience and results . StudiesStudies about the relationship between anxiety and motivation have come up with different conclusions. A large number of scholars hold the belief that anxiety and motivation are negatively correlated. For instance, As discussed above, foreign language anxiety, language learning motivation, and self-efficacy are all important factors in second language acquisition and learning despite learners\u2019 ages, nationalities, and educational backgrounds. Nevertheless, just because of the complexity of learner populations and second language learning process, more and continuous research on these issues are still called for. And although some research shows that they interact with one another and collectively affect second language learning outcomes, little research has examined their interaction simultaneously in the same situation. Moreover, there is hardly any research focusing specifically on top students\u2019 attitudes toward learning a second language and how these attitudes affect their language proficiency. Hence, this research sought to examine English class anxiety, motivation, and self-efficacy in students from various top-notch programs of a top university in China. The following research questions were of particular interest:What are the profiles of the participants in terms of English proficiency, English use anxiety, motivation, and self-efficacy?What is the relationship between the participants\u2019 English use anxiety, motivation, and self-efficacy?How do English use anxiety, motivation, and self-efficacy affect the participants\u2019 standardized test scores?To foster talents and prepare them to be worldwide first-class scholars in basic yet important disciplines such as mathematics and physics, China\u2019s Ministry of Education initiated the Top-Notch Students of Basic Disciplines Training Program in about 15 top universities in 2009. To join the program, students must do well in the National College Entrance Examinations to be admitted by a member university, and then excel in the selective tests of the Top-Notch Program of the university. Each year, the program of each university admits around 200 out of about 4,000 students it enrolls. Students in the program often enjoy more and better resources than other students of the same university. For example, they are more likely to be taught by leading professors and be granted funds for learning and research. Consequently, they generally excel in many aspects and their way of learning is expected to be enlightening to other students. Nevertheless, since 2009 when the program was first launched, not much research has been done on the program or students, even less research in second language acquisition has been done on them. This mainly motivates the present research.The present study was conducted in a highly prestigious state-owned research-oriented university in Beijing , whose TIn total, 223 university students  joined the study, and their average age is 19.67 (SD\u2009=\u20091.067), ranging from 16 to 22\u2009years old. These participants are at different undergraduate years, and they are from various disciplines, including physics, mathematics, chemistry, biological science, life sciences, clinical medicine, computer science, artificial intelligence, interdisciplinary information sciences, theoretical and applied mechanics, foreign language and literature, and philosophy.The participants were required to accomplish a battery of questionnaires see  about thBackground Information (Item 1\u201311): In this section, participants\u2019 age, gender, major, grade, and time length of English use per day were required. Also, participants\u2019 level of English proficiency was recorded in two ways: one was the score of the most recent standardized English test they had taken, for instance, International English Language Testing System (IELTS) or College English Test (CET). The other was the self-rated proficiency of their English speaking, writing, listening, reading, and overall performance, respectively, and each item was reflected on a 1-to-10 scale, with 1 being \u201cnot at all satisfied\u201d and 10 being \u201cvery satisfied.\u201dEnglish Use Anxiety Questionnaire (Item 12\u201319): Motivational Self-Talk Questionnaire (Item 20\u201324): This questionnaire was adapted from that in Self-Efficacy Questionnaire (Item 25\u201327): With reference to the self-efficacy questionnaire used in Language Learning Orientations Questionnaire (Item 28\u201346): The questionnaire used in the current study is based on the Language Learning Orientations Scale of The questionnaires were in Chinese, and they were organized and distributed online to the participants. In total, 223 valid questionnaires were collected and analyzed via SPSS 27. Since the scores of different standard tests were provided under different marking systems (for instance the full marks are 346 for Graduate Record Examination (GRE), 120 for Test of English as a Foreign Language (TOEFL), 9 for International English Language Testing System (IELTS), and 710 for CET-4/6), they were all converted to the centesimal system for computing and analysis. Also, by averaging the scores of related items, the 35 statements of the four latter questionnaires corresponded to 9 variables for analysis: Use of English anxiety , Motivational Self-Talk , Self-Efficacy in learning English , Extrinsic Motivation \u2013 External Regulation , Extrinsic Motivation \u2013 Introjected Regulation , Extrinsic Motivation \u2013 Identified Regulation , Intrinsic Motivation \u2013 Knowledge , Intrinsic Motivation \u2013 Accomplishment , and Intrinsic Motivation \u2013 Stimulation . Means and standard deviations of various items were firstly computed to indicate the general features of the participants, then the relations between different variables were analyzed via correlation analysis and regression analysis.According to the collected data, the participants reported using English for 2.12\u2009h (SD\u2009=\u20091.10) on average per day after entering university. One hundred and ninety-one out of the 223 participants had taken part in a standardized English proficiency test like GRE, TOEFL, IELTS, CET-4, CET-6, Test for English Majors (TEM), and Tsinghua English Proficiency Test (TEPT) (English proficiency test of the university). The scores were then standardized on the scale of 1\u2013100, which showed that the participants had an average score of 80.29 out of 100 (SD\u2009=\u20099.79). Data of self-ratings showed that the participants scored 4.77 to 6.50 in different aspects of English see , indicatp\u2009\u2264\u20090.01). Of all the self-ratings, self-rated proficiency in overall English had the highest coefficient with standardized tests scores (r\u2009=\u20090.487) and that in speaking proficiency (r\u2009=\u20090.810), listening proficiency (r\u2009=\u20090.853), reading proficiency (r\u2009=\u20090.757), and writing proficiency (r\u2009=\u20090.793), respectively. Thus, standardized test scores and self-rated proficiency in overall English were used as indicators of students\u2019 English achievements for further analyses in the present study.Meanwhile, As shown in r\u2009=\u2009\u22120.441, p\u2009\u2264\u20090.01), IMK  and IMS , and it was significantly positively related to IntroR , in which the correlation was moderate with SEQ (0.4\u2009<\u2009|r|\u2009<\u20090.6), weak with IMK (0.2\u2009<\u2009|r|\u2009<\u20090.4), and very weak with IMS and IntroR (0\u2009<\u2009|r|\u2009<\u20090.2). EUAS had no significant correlations with MSTQ, ER, IdenR, and IMA. MSTQ had a significant and positive relation with SEQ  and all six sections of LLOQ , in which the correlation was weak with SEQ, ER, and IntroR (0.2\u2009<\u2009|r|\u2009<\u20090.4) and moderate with IdenR, IMK, IMA, and IMS (0.4\u2009<\u2009|r|\u2009<\u20090.6). SEQ had a significant and positive relation with five sections of LLOQ , except the section of IntroR, which was significantly negatively related , and the correlations were very weak (0\u2009<\u2009|r|\u2009<\u20090.2) both ER and IntroR, weak (0.2\u2009<\u2009|r|\u2009<\u20090.4) with both IMA and IMS, and moderate with IdenR and IMK (0.4\u2009<\u2009|r|\u2009<\u20090.6). At last, all six sections of LLOQ  are significantly positively correlated with each other , with the weakest correlation between IntroR and IdenR (0\u2009<\u2009|r|\u2009=\u20090.179\u2009<\u20090.2) and the strongest between IMK and IMA (0.6\u2009<\u2009|r|\u2009=\u20090.732\u2009<\u20090.8).To explore whether English use anxiety, language learning motivation, and self-efficacy predicted students\u2019 English achievements, multiple stepwise regression analyses were done, with EUAS, SEQ, and IMK being independent variables and standardized test scores and the self-rated overall English proficiency being the dependent variable, respectively. The results are reported in the tables below.\u03b2\u2009=\u20090.268, t\u2009=\u20093.99, p\u2009=\u20090.000, f2\u2009=\u20090.28) and EUAS  were good predictors for the participants\u2019 standardized test scores, with the first being a positive and the second a negative predictor.As shown in \u03b2\u2009=\u2009\u22120.465, t\u2009=\u2009\u22128.200, p\u2009=\u20090.000, f2\u2009=\u20090.07) and SEQ  were good predictors for the participants\u2019 self-rated proficiency in overall English, with the first being a negative and the second a positive predictor.Regression analyses also yielded two models with the change in R2 being 0.357 for model 1 (EUAS) and 0.073 for model 2 (EUAS and SEQ) for self-rated proficiency in overall English. Namely, EUAS  and that of IMK was also significant . Then R2 in step 2 is 0.361 and the dependent variable was SE. Both standardized coefficients of EUAS (\u03b2\u2009=\u2009\u22120.405) and IMK (\u03b2\u2009=\u20090.050) were significant (p\u2009=\u20090.000). At last, all three standardized coefficients were significant when the dependent variable is the standardized test score. The data for each term were SE , EUAS  and IMK . Since the effects of both EUAS and IMK decreased after SE was introduced into the model, the influences of both EUAS and IMK on STS were partially mediated by SE  were significantly negatively correlated was in accordance with the discoveries of multiple previous researchers . SecondlThe regression analyses of how anxiety and motivation affect English achievements were conducted, and the results indicated a significant influence on both the scores of standardized tests and the self-rated proficiency. The effect of learning motivation was positive, and students who were more motivated tended to perform better in language tests and consider themselves more proficient in using English. Oppositely, the impact of English use anxiety could be adverse, interfering the learners\u2019 performance in taking language tests and causing them to underestimate their own language proficiency.English use anxiety and language learning motivation were negatively correlated in the current study, and the correlation was specifically significant between anxiety and intrinsic motivation \u2013 knowledge, as well as between anxiety and intrinsic motivation \u2013 stimulation. Referring to the statements of the questionnaires, English L2 learners who are more anxious with using the language may be feel less willing to joining the course or related activities, which then influences the process of knowledge acquisition. In this way, the intrinsic motivation towards learning new information will be diminished. Also, without enough knowledge learned, the capability of speaking , writingThe mediating effect of self-efficacy was also proved in the study. Firstly, a high level of self-efficacy emphasizes on the learners\u2019 self-confidence in accomplishing the learning assignments and language tests, so that the learners are more determined in their language skills, which can balance the negative effect caused by the anxiety  and leadThe present study explored how English use anxiety, language learning motivation, and self-efficacy were related to one another and how they collaboratively predicted English achievements of students in top-notch programs of a top university in China. Major findings were:The participants had a low-medium level of English use anxiety, and among different types of motivations, extrinsic motivation \u2013 introjected regulation was the least detected (low level), extrinsic motivation \u2013 external motivation and intrinsic motivation \u2013 knowledge was the next in line (low-to-medium level), while extrinsic motivation \u2013 identified regulation, intrinsic motivation \u2013 accomplishment and intrinsic motivation \u2013 stimulation were mostly recognized (medium-to-high level). Also, the participants had a medium-to-high level of motivational self-talk and self-efficacy.English use anxiety and language learning motivation were negatively correlated with each other, and to be more specific, English use anxiety was significantly negatively correlated with both intrinsic motivation \u2013 knowledge and intrinsic motivation \u2013 stimulation.English use anxiety significantly negatively predicted the learners\u2019 English achievements, while intrinsic motivation-knowledge and self-efficacy significantly positively predicted the learners\u2019 standardized test results and self-rated proficiency in overall English, respectively.Self-efficacy of the learners mediated the influence of anxiety and motivation on the top students\u2019 English achievements.These findings further pinpoint the importance of English use anxiety, motivation, and self-efficacy in second language acquisition and learning. They also brought attention to the affect of top students of top universities, focusing on how these leading learners of various disciplines evaluate their language use anxiety, motivation, and self-efficacy. The results showed that for top students in various academic fields, learning and using English still occupies a part of their daily life, and they are mostly motivated by internal determination of accomplishing English-related tasks and the excitement of reaching the goals instead of the external regulations. Consistent with previous research, participants in current studies also experienced negative effects of anxiety and positive influence of motivation on their language achievements, and those with high self-efficacy can deal with the effect better. Besides conveying academic information, L2 teachers should pay more attention to understanding and sensing learners\u2019 attitudes and emotions, and build up efficient regulatory strategies to help the learners cope with tension and stay motivated, so that to reach better performance in both teaching and learning. At the same time, L2 learners, especially the self-taught ones, should be aware of the emotional changes and take active steps to keep a positive attitude toward the learning experience.The current study has some certain limitations. Firstly, the participants took part in different types of standardized tests, and these tests do not always share the same evaluation criteria or level of difficulty. Though the self-evaluation of the participants was also included to provide a more comprehensive image of learner\u2019s language proficiency in the current study, future studies could still discover better ways to describe the participants\u2019 language skills in a more unified manner. Also, this research did not continue to discover how L2 learners cope with their anxiety or develop their motivation, which can be examined by further studies of learning strategies and learning styles. Another aspect that can be included in the following studies is the individual differences among the participants, for instance how English learning has benefited their university life and study of their own majors. These qualitative studies could be realized via interviews or open-ended questionnaires.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.XW: design of study, analysis and interpretation of data, writing-review and editing, and supervision. HY: writing-original draft and analysis and interpretation of data. JL and ZL: data collection. All authors contributed to the article and approved the submitted version.This research was sponsored by the 2021 Top-Notch Students of Basic Disciplines Training Program 2.0 Project (No. 20211008).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.https://www.frontiersin.org/articles/10.3389/fpsyg.2022.953600/full#supplementary-materialThe Supplementary material for this article can be found online at: Click here for additional data file."}
+{"text": "To mitigate parasite resistance to current chemotherapy, computer-aided drug design (CADD) techniques have been employed in the discovery of novel drug targets and the development of small molecule inhibitors to provide an intriguing alternative for malaria treatment. The evolution of plasmepsins, a class of aspartyl acid proteases, has gained tremendous attention in drug discovery, especially the non-food vacuole. They are expressed at multi-stage of the parasite\u2019s life cycle and involve in hepatocytes\u2019 egress, invasion, and dissemination of the parasite within the human host, further highlighting their essentiality. In silico exploration of non-food vacuole plasmepsin, PMIX and PMX unearthed the dual enzymatic inhibitory mechanism of the WM382 and 49c, novel plasmepsin inhibitors presently spearheading the search for potent antimalarial. These inhibitors impose structural compactness on the protease, distorting the characteristic twist motion. Pharmacophore modeling and structure activity of these compounds led to the generation of hits with better affinity and inhibitory prowess towards PMIX and PMX. Despite these headways, the major obstacle in targeting PM is the structural homogeneity among its members and to human Cathepsin D. The incorporation of CADD techniques described in the study at early stages of drug discovery could help in selective inhibition to augment malaria chemotherapy.The past decade has seen most antimalarial drugs lose their clinical potency stemming from parasite resistance. Despite immense efforts by researchers to mitigate this global scourge, a breakthrough is yet to be achieved, as most current malaria chemotherapies suffer the same fate. Though the etiology of parasite resistance is not well understood, the parasite\u2019s complex life has been implicated. A drug-combination therapy with artemisinin as the central drug, artemisinin-based combination therapy (ACT), is currently the preferred malaria chemotherapy in most endemic zones. The emerging concern of parasite resistance to artemisinin, however, has compromised this treatment paradigm. Membrane-bound Ca Plasmodium (P) parasites, which are spread by the bite of female anophele mosquitoes, are responsible for the crippling disease malaria. At least 3.2 billion of the world\u2019s population stand at risk of contracting malaria, and an estimated 350\u2013500 million clinical malaria cases occur annually, with the majority of the incidents emanating from the sub-Sahara regions . In SoutP. falciparum, P. ovale, P. malariae, P. knowlesi, and P. vivax are the five known Plasmodium species to infect humans [P. falciparum is the most common in sub-Saharan Africa and has been implicated in serious life-threatening malaria conditions and thus tagged as the most lethal [Currently, t humans . Among tt lethal . Despitet lethal .Plasmodium species. However, the use of herbs in phytotherapy is often poorly standardized and controlled. Moreover, their active substance content may vary according to plant genetics as well as climatic factors. As such, most malaria control strategies today depend on safe and effective drugs, as they have done for decades [Herbs and herbal extracts are frequently used in various parts of the world to manage and treat malaria because they are affordable, easily accessible, and effective ,6. Curcu decades . The majArtemisinin-based combination therapy (ACT), a drug combination therapy with artemisinin as the central drug, has, over the years, been at the forefront of the treatment of uncomplicated malaria . In the Plasmodium falciparum is prone to drug resistance [Plasmodium vivax is known to put a significant burden on the malaria-endemic world with high morbidity and mortality due to its tendency to cause recurrent infections. Weeks after the initial attack, the dormant liver stages called hypnozoites trigger a relapse of the infection. Recurring infections, which are the primary cause of vivax sickness, can happen as frequently as every three weeks. Chloroquine, the first-line treatment for P. vivax malaria in most endemic countries, faces similar setbacks of resistance in different parts of the world [P. falciparum multidrug resistance gene (pfmdr), which also regulates the resistance phenotype in some PfCRT mutant parasites [dhps-437 and dhps-540 recorded a risk of treatment failure of 37% in children with a parasite load of fewer than 45,000 parasites/\u03bcL [Antimalarial resistance poses a major obstruction to malaria chemotherapy in sub-Saharan Africa  largely sistance . Resistasistance . The parsistance . The firsistance . A summahe world . The traarasites . Folate sites/\u03bcL . ChemicaIn the quest to overcome the global challenge of parasite resistance, researchers have geared prospective studies toward the investigation of novel drug targets whose inhibition is catastrophic to the parasite. Among these targets are plasmepsins, a family of aspartic acid proteases responsible for hemoglobin degradation, hepatocellular egress, and dissemination of the infection within the host. The role of plasmepsin in malaria treatment is further expatiated in P. falciparum. It is, however, intriguing to know that Chloroquine, Amodiaquine, Quinine, and Mefloquine interact significantly with Glutamic acid (Glu) in securing overall stability within the binding pocket. The drugs form a mesh of conventional hydrogen bonds and hydrostatic interaction with Glu, as shown in This chapter elucidates the interaction of quinoline front-line antimalarial drugs in the active pockets of respective target proteins of The hunt for innovative pharmacological targets offering a more alluring path toward the effective treatment and relief of malaria has been prompted by the enormous occurrence of malaria and the rising emergence of parasite resistance to conventional therapies . These tP. falciparum PMIX and PMX against human cathepsin D (4OBZ). The outline of homology modeling techniques is represented in Ligand\u2013receptor complex is essential to envisage the interaction of drug candidates within the catalytic domain of the receptor. These interactions stem from the formation of bonds, most significantly conventional hydrogen bonds and hydrophobic interactions. In drug discovery, drugs are designed complementary to the active pockets. Computation analysis of the ligand\u2013receptor complex encompasses the use of the crystallized structure of the receptor and the ligand. Early studies on the non-food vacuole plasmepsins were centered on homology modeling as the crystalized structures were unavailable at the protein data bank ,19. HomoStructure\u2013Activity Relationship (SAR) is an online technique intended to find relationships between structural-related properties, mostly between chemical structure and biological activities. The pharmacophoric moiety of the therapeutic compound responsible for eliciting biological effects in the target protein is highlighted, serving as a template to search for drug-like compounds with similar structures. This permits adjustment of the effect or the potency of the bioactive compound by altering its chemical structure. This technique was employed in our previous study to identify compounds with the 2,4-diaminopyrimidine rings in the quest to develop inhibitors for the treatment of Alzheimer\u2019s disease . SimilarP. falciparum was carried out [50 of 0.35\u03bcM, 0.4 \u03bcM, and 2.0 \u03bcM, respectively, thus demonstrating the essentiality of this approach in the identification of lead compounds against P. falciparum. Additionally, azithromycin derivatives from SAR showed a significant enhancement in vitro antimalarial activity against P. falciparum; the aromatic moiety on the 9a-position of the sugar-containing macrocyclic ring plays an important role [To forecast new in vitro antimalarial medicines, a quantitative structure\u2013activity relationship analysis employing a library of 395 compounds previously evaluated against chloroquine-susceptible strains of the blood stages of ried out , the stuant role .Virtual screening (VS) is a computational technique engaged in probing libraries of small molecules to identify therapeutic compounds with the tendency to influence drug targets. vs. plays a central role in the early stage of drug discovery and development, one of the productive and cost-effective technology in the search for novel lead compounds . ComparaStructure-based virtual screening (SBVS) is a computational method performed in the early-stage drug discovery process to search libraries of chemical compounds for novel bioactive molecules against a known drug target. SBVS employs a 3D structure of a target protein of interest which is determined experimentally or through homology modeling, as discussed earlier. The active pocket of the target protein is docked by small molecule inhibitors from the database and is ranked based on their predicted binding affinity or complementarity to the binding site. The outcome of SBVS could be hundreds of compounds, but mostly only a few top-ranked drug-like compounds are selected for further experimental assays. This approach is essential in the identification of hits.Ligand-based virtual screening technique employs the therapeutic information present in a known active compound rather than the structure of a target protein for the identification and optimization of lead. To find potentially active drug-like molecules for experimental evaluation, large biodata sets are investigated. Quantitative structure\u2013activity relationships (QSAR), pharmacophore mapping, similarity substructure searching, and machine learning are examples of methods used in LBVS. Ligand-based visual screening of dual PMIX and PMX inhibitor 49c led to the identification of three leads from the ZINC database, namely; ZINC03351002, ZINC03417506, and ZINC02619047 [The drug-likeness assessment of potential pharmaceutical compounds is an important stage in the drug discovery process. It evaluates the administration, distribution, metabolism, excretion, and toxicity of the compound of interest. The Lipinski rule of five is an online tool to predict the drug-likeness of a compound with parameters such as molecular weight, blood-brain permeability, gastrointestinal absorption, and lipophilicity, among others. This technique has been employed in previous studies ,21 in thThis computational technique mimics the interaction of ligand\u2013receptor complexes at the molecular level as though it is in a living system. MD simulations generate trajectories that are saved at 1 ps. Post-dynamic analysis on each of the systems under study is performed using the CPPTRAJ module within the AMBER 14/18 package. Subsequent graphical representations are conducted with the aid of the user interfaces of UCSF chimera and Microcal Origin analytical tools for the evaluation of structural dynamics such as the root mean square deviation (RMSD), root mean standard fluctuation (RMSF), and thermodynamic energy evaluation .Recognition of plasmepsins in antimalarial drug discovery, though renowned, is yet to unknot an approved drug. Several proposed inhibitors were abandoned due to the limited potential of their therapeutic pathway. PMs are characterized by \u201ctwo\u201d aspartic acid residues in the active site. Presently there are ten classes of plasmepsins; PM I-X . PlasmepPepstatin A showed aspartic protease inhibitory prowess, specifically against PMII. However, the adverse side effect upon the administration of Pepstatin A renders its clinical usage questionable. Small molecule competitive inhibitors 49c and WM382 have experimentally proven to exert dual inhibitory prowess on both PMIX and PMX [Hexapeptide statin-containing amino acid  and PMX ; the enzP. falciparum is the only species that has numerous plasmepsins active within the food vacuole. The development of the intraerythrocytic stages of malarial parasites depends on hemoglobin (Hb) breakdown. Four aspartic proteases, PMI, PMII, Histo-aspartic acid (PMIII), and PMIV, are known to be involved in this process, which takes place inside an acidic digestive vacuole (DV) [ole (DV) . These eole (DV) .Pepstatin A, with a putative catalytic dyad comprising histidine and aspartic acid [Hemoglobin degradation involves multiple stages whose initiation is specifically conducted by PMI and PMII. These two proteases cleave the native hemoglobin tetramer 54 between \u03b1Phe33-\u03b1Leu34 in the highly conserved hinge region, unraveling and exposing the protein to further degradation. This collective effort may enable efficient degradation of the molecule. Plasmepsin III, also known as HAP, is an active protease with a catalytic mechanism activity that mimics that of the aspartic acid proteases in hemoglobin degradation and seemingly has a high affinity for The remaining members of the plasmepsin family PMV-PMIX are known as non-food vacuole plasmepsin as they are effective outside the acid food vacuole. Unlike the FV plasmepsins, this class of plasmepsins shows relatively lower structural similarity. Sequence alignment analysis of PMIX and PMX showed a structural similarity of 44.31% (unpublished paper). In comparison, PMVI-VIII are expressed in the vector during the parasite\u2019s intraerythrocytic stages of sporozoite formation and motility. They, however, do not play a direct role in the transmission of the parasite within the human host [The food vacuole class of plasmepsins has gained much attention, yet there is no approved drug in this category . Recent Another headway inhibitor is compound 49c, similar to WM382, 49c causes structural compactness on both proteases, thereby prohibiting the characteristic twist motion of the flap residues ,19. UsinThe characteristic prowess of plasmepsin inhibitors to impose structural compactness on their target proteins restricts the typical twist motion, especially in the flap and hinge regions. The flap regions usually move away from the binding site exposing the catalytic dyad of the protease to the ligand. The chemical structures of compounds WM382 and 49c are represented in Pepstatin A, a hexapeptide statin-containing amino acid, despite its inhibitory prowess against the food vacuole plasmepsins, was associated with off-targeting of the host Cathepsin D [The development of highly selective plasmepsin inhibitors remains a major obstacle in the drug discovery and development domain, as these proteases exhibit a high percentile of structural homogeneity. PMIX and PMX share over 40% structural similarity, including the catalytic dyad. Aside from the intra-family similarities, there is an alarming concern about the degree of the monotony of plasmepsins to human Cathepsin D, a pervasive lysosomal enzyme present in most cell types. The undesirable effect of hepsin D ,38.The lack of competitive inhibitors and the increasing challenge of parasite resistance to current antimalarial drugs has triggered the search for more intriguing drug targets and small molecule inhibitors to augment malaria chemotherapy. This current study discussed antimalarial drugs and transport proteins implicated in their resistance. Furthermore, we elaborated on the emergence of aspartic acid proteases as potential drug targets in the discovery and development of antimalarial drugs. In silico and bioinformatic techniques to speed up the antimalarial drug discovery and development to mitigate parasite resistance in malarial chemotherapy were as well expounded.Pepstatin A. To cover these challenges, computed-aided drug design (CADD) techniques have been incorporated. These in silico approaches aid in evaluating the pharmacokinetics and potency of drug candidates. Moreover, these methods predict the binding orientation and affinity of drug candidates to their respective targets, thereby reducing the incidents of off-targeting. Structure\u2013activity relation and ligand/structure-based virtual screening augment the discovery of new drug candidates driven by the pharmacophoric moieties of known drugs or experimental inhibitors. Through molecular dynamic simulation, the enzyme inhibitory mechanism of potential drug candidates could be demonstrated and predict their usefulness in disease control.Hemoglobin degradation, hepatocellular egress, and dissemination of malaria parasites within the host are significant factors that influence the incision and virulence of malaria infection. Arresting the perpetrators of these acts is a step toward the relief and treatment of malaria. 49c and WM382, and experimentally proven small molecule inhibitors, are spearheading the course of plasmepsin inhibition. Both compounds have dual inhibitory activities against PMIX and PMX, the two vital non-food vacuole plasmepsins. 49c and WM382 form stable conformation within the active pocket of the proteases, which is initiated by hydrogen and hydrophobic interaction with the catalytic dyad and cause rigidity of the protease. Despite the advancement, the obstacle, however, is the structural homogeneity expressed among the family of plasmepsin as well as their similarity to human Cathepsin D, which hindered the clinical administration of"}
+{"text": "Burnout is a growing problem among medical staff worldwide and empathy has been described as an essential competence to attenuate burnout. Previous studies found job satisfaction and job commitment were affected by the empathy and associated with burnout. This study explores the effect and mechanism of empathy on burnout on medical staff and investigates the mediating role of job satisfaction and job commitment in the relationship between empathy and burnout among medical staff.Based on a self-administered questionnaire which included the Maslach Burnout Inventory (MBI) to measure burnout, 335 responses from medical staff in Tianjin City, China, yielded data on socio-demographic characteristics, empathy, burnout, job satisfaction and job commitment. Bivariate correlation and structured equation modeling (SEM) analyzed the relationships between empathy, job satisfaction, job commitment and burnout multi-group invariant analysis was used to evaluate whether the model was consistent across different type and level of hospitals and different job and employment type subgroups.p\u2009>\u20090.05) and hospital type (p\u2009>\u20090.05), but was inconsistent across hospital level (p\u2009<\u20090.05) and job type (p\u2009<\u20090.05).A total of 202 (60.3%) medical staff had low level burnout, 115 (34.3%) staff had the moderate level and 18 (5.4%) staff had the high level burnout. The results of the SEM showed that empathy not only had a direct negative effect on burnout (Our results pointed out the important role that empathy plays in addressing burnout and revealed that managing job satisfaction and increasing the job commitment attenuated burnout. We recommend that the government should accelerate the reform of the resourcing of different hospital levels; facilitate hospital managers to implement additional training; and support hospitals to strengthen psychological testing and counseling to reduce medical staff burnout.The online version contains supplementary material available at 10.1186/s12889-022-13405-4. Burnout, or when individuals experience chronic work-based stress for which the perceived job demands exceed personal and work-place resources, is common among medical workers . BurnoutH1: Empathy has a negative association with burnout.The health care literature has explored the factors attenuating medical staff burnout, including empathy, job commitment and job satisfaction. Empathy is a unique psychological resource because of its impact both on patients and medical staff. Through the formation of personal relationships and enhanced communication with patients , empathyH2a: Empathy has a positive effect on job satisfaction.H2b: Job satisfaction negatively mediates the relationship between empathy and job burnout.H3a: Empathy has a positive effect on job commitmentH3b: Job commitment negatively mediates the relationship between empathy and job burnoutIn the burnout literature, many factors have been associated with burnout in medical staff, including sex, working hours, the presence of work-home conflict, coping strategies, staff perceptions, job commitment and job satisfaction . Job satBased on the above discussion, Fig.\u00a0Previous research also found that the empathy ability and burnout level are different through several personal characteristics, such as work position, working experience and hospital types , 21, 40.H4: The model in Fig.\u00a0Our study extends the empathy-burnout literature in several ways. Previous studies have mainly focused on the relationship between job burnout and empathy, or job commitment and job satisfaction and burnout. Few studies have focused on the mediating effect of variables on the empathy-burnout relationship. Constructing a mediated structural equation model, we extend the burnout literature by assessing whether job satisfaction and job commitment mediated the relationship between empathy and job burnout. Second, most burnout studies have collected data on one type of health professional, such as doctors or nurses, in a specific health setting, such as nursing homes or hospitals . Our stuUsing cluster sampling, we conducted a pilot survey to assess the robustness and validity of our variables and the framing of our questions, before undertaking face-to-face and online surveys of medical professionals from six hospitals in Tianjin, China from July 1, 2019 to September 8, 2019. After the agreement of the human resource office in each hospital, we trained the staff on how to administer the questionnaire, the understanding of informed consent and the meaning of each questionnaire item. The questionaries were accompanied by an official letter from the human resource office in each hospital. Respondents could scan the QR code on their mobile phones to complete the survey or fill out paper questionnaires. In order to improve the accuracy of the survey responses, the survey purpose, method and the conceptual basis of each variable was explained to participants. The inclusion criteria of participants captured physicians, nurses, medical technicians, pharmacists, administrators and other medical personnel with more than 1\u00a0year of work experience engaged in medical and health work in Tianjin public medical and health institutions. All participants were volunteers, who gave informed consent to participate and were capable of completing the questionnaire independently. The responses by participants were anonymous.The minimum sample size was calculated as 270 completed surveys \u201344. Of tThe questionnaire composed five parts: job burnout, empathy, job satisfaction, job commitment and the respondent\u2019s social-demographic characteristics, comprising age, sex, educational background, marital status, salary, workplace, professional titles and average daily working hours. The content of each scale is presented in Supplementary Table To measure burnout, we employed the most commonly used burnout scale, the Maslach Burnout Inventory (MBI) , 45. ComEmpathy of medical staff was measured by the Chinese version of the Jefferson Scale Empathy \u2013 Health Professions (JSE-HP) . The 20-Adopted from Zhang et al. , 49, theTo measure job commitment, we used Meyer's three-dimensional scale, comprising \u2018Emotional Commitment\u2019(EM), \u2018Continuous Commitment\u2019 (CC) and \u2018Normative Commitment\u2019 (NC) items, and measured by a 5-point Likert scale (1\u2014very dissatisfied to 5\u2014very satisfied).Cronbach's alpha values of each dimension in each scale and the values of the overall scale were all greater than 0.7, indicating good factor reliability. Most of the average variance extracted (AVE) values were greater than 0.5, indicating good aggregation validity of the model . The detStructure equation modelling (SEM) refers to equations using parameters in the analysis of the observable, such as income or working hours, and latent variables, such as satisfaction and emotion. This study constructs a structural equation model of empathy and job burnout in medical staff and uses path analysis and diagrams to explain the internal relationships among variables to verify our hypotheses.Mediating variables could explain the process of \u201chow\u201d and \u201cwhy\u201d between two variables. The influence of independent variables on dependent variables has both direct effects and indirect effects through mediating variables. Our research analyses and verifies the mediating effect of medical staff\u2019s job satisfaction and job commitment between empathy and job burnout.This method was used to test whether the model we built is suitable for different population groups, which is whether the model is invariant among different sample groups. Additionally, it could verify whether a certain variable will have a moderating effect on the model. To test multi-group invariance, we divided our sample into three different hospital levels ; two hospital types ; three job positions ; and two employment types (authorized employees and unauthorized employees).Data entry and conversion was completed with EpiData 3.1, and SPSS 24.0  and AMOS 23.0  was used to analyze the data. Descriptive analyses explored the key social demographic factors and the degree of job burnout of medical staff. Next, a structural equation model (SEM) was specified to analyze empathy and job burnout, mediated by job satisfaction and job commitment. Based on the SEM, a multi-group equivalence analysis was conducted to determine the impact of different types and levels of hospital, job positions and employment types on the model. In the process of model fitting, 5000 Bootstrap tests to the mediating variables were conducted; observed variables with path coefficients less than 0.6 were deleted; and the modification index (MI) was used to optimize the model.The socio-demographics of the 335 medical staff who completed the survey are presented in Table There were 45 (13.4%) staff with low level burnout, 82 (24.48%) staff with moderate level burnout and 46 (13.73%) staff with high level burnout. Table p\u2009<\u20090.01), and the highest correlation was emotional exhaustion (r\u2009=\u20090.933).Table p\u2009<\u20090.01, r\u2009<\u20090) and positively correlated with job commitment . Job burnout was also significantly negatively correlated to job commitment and job satisfaction  and job commitment and job satisfaction were negatively correlated. .Table According to Fig.\u00a0df\u2009=\u20092.80, AGFI\u2009=\u20090.914, GFI\u2009=\u20090.937, CFI\u2009=\u20090.986, TLI\u2009=\u20090.964, RMSEA\u2009=\u20090.073, all of which meet the AMOS reference criteria [We revised the model when some of the fitting indices did not meet the fitting criteria, indicating that the path map Fig.\u00a0criteria . The coeAs shown in Fig.\u00a0p<0.01), 95% CI: (-0.159) - (-0.041), which was opposite to H2b. Job commitment had a negative mediation effect (p<0.01), 95% CI: (-0.544) - (0.269), which supports H3b.Each path of the mediating variables was guided by 5000 repetitions of the Bias-corrected Bootstrap test and Percentile Bootstrap test. Supplementary Table The multi-group invariance analysis tests whether the model is consistent across different subgroups, comprising different types of hospital, different levels of hospital, different medical jobs and different employment types. When there are (in)significant differences between different subgroup sample coefficients, the research model parameters are (in)variant, indicating that the multi-group model is (in)consistent across different subsamples.p\u2009<\u20090.05). With regard to job satisfaction\u2014job commitment path, the tertiary hospitals and secondary hospitals differed significantly compared to primary hospitals (p\u2009<\u20090.001). As shown in Supplementary Table p\u2009<\u20090.05), with the influence of the secondary hospital significantly greater than that of primary hospitals.Based on primary, secondary and tertiary hospital level, the results show that hospital level had a significant impact on the empathy and job burnout (p\u2009<\u20090.05). Supplementary Table p\u2009<\u20090.01) and between nurses and other medical staff (p\u2009<\u20090.05). The nurse group had a significant impact on the prediction of job burnout (p\u2009<\u20090.001), while the physician group impact was not significant. The model showed that the influence of nurses was greater than that of physicians and other medical staff. On the job commitment\u2014job burnout path, there was a significant difference between physicians and nurses (p\u2009<\u20090.001), between nurses and other medical staff (p\u2009<\u20090.05), and between physicians and other medical staff (p\u2009<\u20090.001). The influence of nurses was greater than that of physicians and other medical staff on the empathy\u2014job commitment (p\u2009<\u20090.05). Further, the job satisfaction of physicians (p\u2009<\u20090.05) and other medical staff (p\u2009<\u20090.001) had a greater impact on job commitment than nurses\u2019 job satisfaction, and other medical staff had more influence than physicians on job commitment (p\u2009<\u20090.001).Next, we tested whether medical job type impacted empathy, job commitment and job satisfaction by dividing the sample into three medical occupation types: physician, nurse and other medical staff. The multi-group nested model indicated that different job type had a significant impact on empathy and job burnout model  revealed consistency across different employment types, which showed the empathy and job burnout model were not affected by employment type. Finally, hospitals were divided into two types, specialty hospitals and general hospitals. The structure weight model test showed a good fit , which means that the empathy and job burnout model was not affected by hospital type. These results show that the model was consistent across employment types and hospital type, which partly confirms H4. The model was not consistent across hospital level and job type.We also explored whether the employment type of medical staff impacted the model. Medical staff were divided into authorized and unauthorized employment types. First, the structural equation model ; higher than the French moderate level burnout (17.8%); and much higher than the French high-level burnout (4.6%)  and consWe found significant differences in burnout according to age, marital status and working hours. Medical staff aged 56-66 years had the highest levels of job burnout. This finding is consistent with the argument that burnout is a gradual process, with the decline of job freshness and the increase of job challenges and difficulties, making medical staff more prone to burnout . But thep<0.01), which is consistent with H1 and previous research [The bivariate spearman correlation analysis of job burnout, empathy, job satisfaction and job commitment showed empathy was negatively correlated with job burnout , which is consistent with the previous research [p < 0.01), where increasing the job commitment of medical staff can shield health workers from burnout [The structural equation modelling tested for direct and indirect path effects. Empathy was positively correlated with job commitment , while primary hospitals were not (p\u2009<\u20090.001). The heavy workload in primary hospitals, with a shortage of staff and the lack of social security and training opportunities, led to a decline in management efficiency and low service capabilities [Regarding the results of multi-group invariance analysis, hospital level and job position played a moderating role in our model. We found that the job satisfaction\u2014job commitment and job satisfaction\u2014job burnout paths for tertiary hospitals and secondary hospitals were significantly different (bilities , so medibilities .The predicted effect of empathy on job burnout and job commitment on nurses was greater than that on physicians and other medical staff, which is consistent with previous research . EmpathyOur results found that employment type and hospital types did not influence the empathy\u2014job burnout relationship, which indicates cross-group consistency. There are few previous SEM studies on the relationship between employment type and hospital types and empathy\u2014job burnout. One study of Chinese medical staff found that authorized staff were more likely to suffer burnout from more work and higher requirements than the unauthorized staff . HoweverOur results suggest that the government should increase the training of medical staff, paying special attention to the training of staff in primary health care facilities , 35. As Hospital managers should enhance the job commitment of their medical staff by creating a good working environment, carrying out progressive education and giving medical staff training and learning opportunities. For example, advanced-study and experience exchange meetings would strengthen staff professional and technical levels, enhancing an individual\u2019s coping ability by giving medical staff self-confidence to solve work problems. Higher levels of professional and technical ability would also promote job commitment. Through the establishment of a scientific shift-work systems, hospital managers should set the maximum daily working hours to avoid excess work hours and reduce the workload. At the same time, hospital managers can promote activities between different departments to create the formation of good cooperative relations among medical staff to improve their empathy ability.Three limitations of the study should be addressed. First, although the SEM was used to quantitatively verify the relationship between variables, the use of cross-section data limited drawing causality conclusions. We also note that burnout might influence empathy, which requires long-term time series data and formal modelling as future research steps. Second, we only surveyed medical staff in Tianjin, and generalizing our results for other cities and regions, especially rural areas, requires further study. Third, our findings are largely based on the self-assessment questionnaires, with the risk that the participants\u2019 answers might have been affected by response and social desirability bias.This study found that different types of medical staff in Tianjin experienced low level burnout rates between and 36 and 76%, moderate level burnout rates ranging between 20 to 43.5% and high-level burnout rates ranging from 4.0 to 8.7%. Our results revealed that empathy was negatively associated with burnout and job commitment had the negative effect on burnout. Job satisfaction had the positive effect on burnout, which was contrary to our hypothesis. Job satisfaction and job commitment played a mediating role in the relationship between empathy and burnout, which were strongly inter-correlated. We found the significant differences in burnout according to age, marital status and working hours. Additionally, the results indicated that the model was consistent across employment type and hospital type but was inconsistent across hospital level and job type. Based on our results, we recommend accelerating the reform of resourcing different hospital levels; facilitate hospital managers to implement additional training; and support hospitals to strengthen psychological testing and counselling.Additional file 1:Supplementary Table 1. Contents of three scales in the questionnaire. Supplementary Table 2. The reliability test results. Supplementary Table 3. The Person correlation results and AVE square root values. Supplementary Table 4. Mediating effect test analysis of paths by bootstrap. Supplementary Table 5. The path coefficient analysis of hospital levels. Supplementary Table 6. The path coefficient analysis of job position."}
+{"text": "In the current healthcare environment, job burnout among medical staff is increasingly evident. Burnout not only affects the mental and physical health and career development of individuals but also affects the quality of care and the doctor\u2013patient relationship. This paper investigates the influence of sports involvement on burnout in medical staff based on the job demands\u2013resources theory, focusing on the mediating role of health anxiety and self-efficacy in the relationship between sports involvement and job burnout. A questionnaire survey was used to collect data from 444 medical staff in public hospitals in Wuhan, China. Structural equation modeling (SEM) with a bootstrapping approach was conducted to test the hypothesis and mediating effects. It was found that health anxiety and self-efficacy played a significant mediating role between sports involvement and job burnout. The results indicate the important role that sports involvement plays in addressing burnout, revealing that decreasing health anxiety and increasing self-efficacy attenuated job burnout. This finding suggests that hospital administrators should not only pay attention to medical staff\u2019s health conditions and improve their enthusiasm for work but also encourage them to become more engaged in sports. During the past two decades, the world has experienced outbreaks of several highly infectious diseases and serious public health emergencies. Expecting to experience increases in workload, medical staff also face problems such as conflicts between professional obligations and personal safety, coupled with high stress, infrequent interpersonal contact, irregular work, and not enough time to rest. Long-term exposure to these stressful situations can have certain effects on the physiological, psychological, and social health of medical staff, leading to hypertension, anxiety and depression; this, in turn, affects work efficiency, which may eventually lead to job burnout ,2. AfterHealth anxiety is a condition characterized by excessive preoccupation with an individual\u2019s health , definedSelf-efficacy refers to the subjective prediction of one\u2019s ability to accomplish a certain task, as well as the individual\u2019s tendentious judgment and feeling on whether one\u2019s behavior can achieve a certain goal . It has The following sections are structured as follows: Demerouti and Bakker et al.  proposedUnder the normal of epidemic prevention and control, the medical industry is an important guarantee to avoid infection and ensure people\u2019s lives and health, and the medical staff is constantly exposed to high stress due to the high risk of infection, isolation, caring for critically ill patients, and overwork. This difficult situation leads to mental health problems such as anxiety, fear, stress, insomnia, and depression . The croPeople with higher self-efficacy can complete the original sports involvement plan when they confront difficulties ; self-efHypothesis\u00a01\u00a0(H1).Sports involvement is negatively associated with health anxiety.Hypothesis\u00a02\u00a0(H2).Health anxiety is positively associated with job burnout.Hypothesis\u00a03\u00a0(H3).Health anxiety mediates a relationship between sports involvement and job burnout of medical staff.Hypothesis\u00a04\u00a0(H4).Sports involvement is positively associated with self-efficacy.Hypothesis\u00a05\u00a0(H5).Self-efficacy is negatively associated with job burnout.Hypothesis\u00a06\u00a0(H6).Self-efficacy mediates a relationship between sports involvement and job burnout of medical staff.Considering these arguments on the relationship between sports involvement, health anxiety, self-efficacy, and job burnout, the conceptual model of the present paper to examine the research hypotheses is presented in The researchers adopted the snowball sampling method, a sample is the participants you select from a target population to make generalizations about. The researchers selected medical staff from a three-tier public hospital in Wuhan. With the support of schools and hospitals, questionnaires were distributed by heads of departments to medical staff in their departments. The questionnaire was conducted from April to May 2022. Before the date of collection, the researchers conducted a pilot test with a sample of 50 medical staff from a hospital to assess the validity of our variables and the framing of questions. All participants voluntarily participated in the survey and earned a gift worth about 5 USD as a response incentive. The responses by participants were anonymous. Out of 500 distributed questionnaires, 56 uncompleted and inaccurate questionnaires were excluded, providing 444 valid responses, with an 88.8% effective response rate.To measure sports involvement, three items were extracted from the research of Beaton et al. . The scaStructural equation modeling (SEM) with AMOS 26.0  was used to examine the hypothesized model . The resCronbach\u2019s \u03b1 coefficient (C\u03b1) and the composite reliability (CR) coefficient of the latent variables, as an acceptable approach proposed by Fornell and Larcker , were caPearson\u2019s correlation coefficient depicts the extent that a change in one variable affects another variable. This relationship is measured by calculating the slope of the variables\u2019 linear regression. Researchers verify the discriminative validity of the data by comparing the correlation coefficient of each variable with the square root of the AVE. 2/df) = 2.146, the goodness of fit (GFI) = 0.951, the normed fit index (NFI) = 0.942, the comparative fit index (CFI) = 0.968, the Tucker\u2013Lewis index (TLI) = 0.959, and the root mean square error of approximation (RMSEA) = 0.051, with reference to the suggested value of Joseph F. Hair et al. [\u03b2 = \u22120.234, p < 0.001) and self-efficacy , supporting H1 and H4. The effect of health anxiety on job burnout was statistically significant , supporting H2. The effect of self-efficacy on job burnout was statistically significant , supporting H5.The structural modeling results indicated that the structured model has a good fit: the chi-square of degrees of freedom , which provides support to H3. It also showed that mediation occurred between sports involvement and job burnout through self-efficacy , which provides support to H6. The findings indicate that medical staff who have more sports involvement, higher self-efficacy, and less health anxiety are less likely to develop job burnout, and the two mediating variables of health anxiety and self-efficacy explained 18.3% of the variance in job burnout.This study followed the recommendation of Bollen and Stine  and usedThis study has made the following contribution to research on job burnout. First, it clarified the positive impact of sports involvement on alleviating individual job burnout by investigating burnout and sports involvement among 444 medical staff and further explored the specific mechanisms of the impact of sports involvement on job burnout by adding two psychological variables\u2014health anxiety and self-efficacy\u2014as mediating variables. The results indicated that the positive effect of sports involvement on job burnout was fully mediated by health anxiety and self-efficacy, and the two mediating variables of health anxiety and self-efficacy explained 18.3% of the variance in job burnout. According to previous studies, job burnout has been defined as a long-term reflection of emotions and stress at work and a negative work-related state of mind as a comprehensive indirect reflection of physical, mental, and emotional aspects. From this perspective, both emotional exhaustion and negative evaluations of oneself can affect an individual\u2019s attitude toward work. Health anxiety is an excessive focus on the fear of illness or a strong belief in illness, and some people suffering from health anxiety will experience some physical symptoms of chest pain, abdominal pain, facial pain, or headaches. Essentially, it exerts an influence on job burnout by changing the individual\u2019s emotional exhaustion of burnout. Studies have been conducted linking excessive physical fatigue to emotional exhaustion. Self-efficacy, however, influenced job burnout by changing one\u2019s positive self-perceptions. From previous studies, self-efficacy was found to significantly and negatively predict the level of job burnout. The higher the level of self-efficacy for medical staff, the lower the level of job burnout in the same professional environment and the more they believe they can handle occupational stress and have the ability to perform their jobs when faced with stress and professional dilemmas. In contrast, medical staff with low self-efficacy find it difficult to adjust their state from occupational distress and tend to consume more energy and approach their work with a negative attitude, which eventually aggravates the degree of job burnout and triggers a series of physical and mental health problems.Second, this paper is the first attempt to explain the effects of sports involvement on health anxiety, self-efficacy, and job burnout using the JD-R theory. JD-R theory explores the effects of job characteristics on job burnout in terms of job demands and job resources, and previous studies have focused on opportunity or resources factors at the physical, social, and psychological levels ,51; in cIt has been shown that the high-pressure and high-load working environment, and the high-risk occupational characteristics of medical staff consume their energy, resulting in a high incidence of job burnout. Long-term burnout not only depletes the physical and mental health of medical staff but also reduces the quality of the healthcare services provided and leads to conflicts between doctors and patients . This stConsidering the impact of sports involvement on health anxiety, self-efficacy, and job burnout, this study makes the following suggestions for medical staff. First, more attention should be paid to strengthening medical staff awareness of the concept of lifelong sports. The government and hospital managers should incorporate sports exercise venues for medical staff into the overall planning of hospital development, regularly hold various lectures on sports topics, propagate the skills and methods of scientific exercise, and carry out more sports activities, such as jumping rope, table tennis, and other competitive projects, to guide medical staff to actively exercise. Second, more scientific exercise programs should be pursued. Hospitals should rely on sports managers to customize personal sports prescriptions for medical staff to provide guidance on when to exercise and to have appropriate knowledge and information about sports at the same time as exercise.In response to the proposed research objectives, this study is motivated by the widespread phenomenon of burnout among medical staff in China. Medical staff is faced with a heavy workload, tedious work tasks, and long working hours, and their common responsibilities involve the assessment, planning, and administration of daily patient treatment and health management. They are more likely to suffer from health disorders and experience symptoms such as neck and back pain, fatigue, and sleep disturbance that may cause anxiety and depression, leading to burnout. Moreover, the result demonstrates that health anxiety and self-efficacy are important factors that affect medical staff sports involvement and job burnout. In particular, sports involvement affects job burnout indirectly through the mediating effects of health anxiety and self-efficacy. Therefore, this study recommended that hospital managers pay attention to medical staff\u2019s health conditions, improve their work motivation and encourage them to be more engaged in sports.This study has certain limitations. First, the study design was cross-sectional. Thus, the relationship between sports involvement, health anxiety, self-efficacy, and job burnout cannot be assumed to be causal, and longitudinal research is necessary to examine the causal relationships in this study for the future. Second, we adopted the snowball sampling method; thus, the reported information may not accurately reflect the underlying values of each variable, unlike random sampling. Third, the two mediating variables of health anxiety and self-efficacy explained 18.3% of the variance in job burnout, but there remains room for a lot of unexplained variation. Further research can provide more possible variables based on this."}
+{"text": "Background: Three-dimensional (3D) printing is an emerging tool in the creation of anatomical models for surgical training. Its use in endoscopic sinus surgery (ESS) has been limited because of the difficulty in replicating the anatomical details.Aim: To describe the development of a patient-specific 3D printed multi-material simulator for use in ESS, and to validate it as a training tool among a group of residents and experts in ear-nose-throat (ENT) surgery.Methods: Advanced material jetting 3D printing technology was used to produce both soft tissues and bony structures of the simulator to increase anatomical realism and tactile feedback of the model. A total of 3 ENT residents and 9 ENT specialists were recruited to perform both non-destructive tasks and ESS steps on the model. The anatomical fidelity and the usefulness of the simulator in ESS training were evaluated through specific questionnaires.Results: The tasks were accomplished by 100% of participants and the survey showed overall high scores both for anatomy fidelity and usefulness in training. Dacryocystorhinostomy, medial antrostomy, and turbinectomy were rated as accurately replicable on the simulator by 75% of participants. Positive scores were obtained also for ethmoidectomy and DRAF procedures, while the replication of sphenoidotomy received neutral ratings by half of the participants.Conclusion: This study demonstrates that a 3D printed multi-material model of the sino-nasal anatomy can be generated with a high level of anatomical accuracy and haptic response. This technology has the potential to be useful in surgical training as an alternative or complementary tool to cadaveric dissection. Endoscopic sinus surgery (ESS) represents the current gold standard technique for several pathologies, ranging from chronic rhinosinusitis to benign and malignant neoplasms . The parThese peculiarities account for the importance of specific training for this type of surgery, aimed to both comprehensively understand sino-nasal anatomy and develop the required surgical skills in a risk-free environment, before operating on a patient .The restrictions of traditional training based on cadaveric dissections have led to the investigation of alternative training methods, including the use of synthetic anatomical models created with 3-dimensional (3D) printing. The emerging advanced additive manufacturing technologies have lately expanded the possibilities of representing 3D patient-specific anatomies, which are extremely useful for both surgical planning, training  and mediThe aim of this study was to develop a patient-specific simulator for ESS using the 3D printing multi-material jetting technology, which reproduces both soft tissues and bony structures, thus increasing the anatomical realism and tactile feedback of the model. Moreover, a preliminary validation of this simulator as a training tool for ESS among a group of ear-nose-throat (ENT) residents and experts, is described.The design and fabrication of the simulator consisted of three steps: A) Image segmentation and 3D modeling of the anatomical components; B) Design of the simulator box and the external face mold; C) 3D printing of the simulator components.The simulator was then assembled and tested by the surgeons in the operation room.TM , which is a certified software for medical 3D modeling. The anatomical structures of interest, i.e., the midface bony structures, the turbinates, and nasal septum were segmented in collaboration with a radiologist with expertise in head and neck anatomy. The segmented mask for the turbinates was slightly reduced, in accordance with the surgical team, to simulate the effect of decongestants used before the ESS. 3D meshes were then generated from all the segmented masks and saved in STL format; For converting the segmentation mask to 3D mesh, we used the standard settings provided by D2P software. In detail, these settings include selection of: \u201cMaintain original morphology\u201d method  and selection of \u201cSimplify and smooth\u201d (to decimate and apply extra smoothing to the mesh surface).The preoperative computed tomography (CT) scan of a patient with no evidence of sino-nasal pathologies was acquired. The Digital Imaging and Communications in Medicine (DICOM) data were imported into D2PThe meshes were imported and processed into 3-Matic Medical software : all the structures were cut coronally into two sections to ease the cleaning of the internal parts after the printing process. According to the surgeons\u2019 requests, the medial periorbital surface and the dura mater of the anterior skull base were selected as relevant anatomical landmarks for surgery, and they were enlarged in order to be more visible in the simulator. Moreover, the nasolacrimal duct and the lacrimal sac were reproduced, filling the segmented bony duct and the lacrimal sac fossa , 2. FinaAs final step, the Fix Wizard tool of 3-Matic software was applied to the meshes to make sure they were printable.A box including the reconstructed patient-specific paranasal sinuses model was created using the open-source software FreeCAD. The box was designed as a hollow rectangle combined with a larger base in order to secure it to the operating table during the simulation.TM, then converted into a mesh and exported as STL file to be further processed in MeshMixer software .The portion of the facial skin was segmented in D2PUsing the negative of the facial skin model, a rectangular mold was designed to allow the subsequent silicone casting .Lastly, the box was provided with a closing lid that was designed in FreeCAD. The patient-specific portion of skin was combined with the lid to let its borders assume the shape of the patient-specific face.The patient-specific model of the paranasal sinuses was printed using the J720 Dental 3D printer . This printer is based on material jetting technology, a process that resembles inkjet paper printing since the material is dropped through small diameter nozzles. In this case, the printing material is a photopolymeric resin subsequently hardened by a UV lamp.via a mixing of rigid resins (VeroMagenta) and a rubber-like material from the Polyjet photopolymer family (Agilus30) with a resulting Shore-A 50. Also, the membranes replicating the orbital content and the dura mater as well as the middle, the superior turbinates, and the inferior turbinates were printed combining Agilus30 and rigid resin VeroMagenta with Shore-A 50 and Shore-A 30 respectively. The lacrimal sac was printed by mixing VeroYellow and Agilus30 with a Shore-A 35 , which was previously used for reproducing bones in other 3D printed phantoms of thorax, pelvis and prosthetic fingers . Consideore-A 35 . ParticuRegarding the other components of the simulator, the box and the lid were both printed using a stereolithography (SLA) 3D printer  with White resin.The components of the patient-specific facial skin mold were also printed using Form 3 SLA printer but with a Clear resin so that, during the silicone injection, both the level of silicone and the presence of air bubbles could be seen.The patient-specific facial skin was realized by a silicone casting through the mold: the selected silicone was Dragon Skin FX-Pro  which is a bi-component silicone frequently used in literature to reproduce human skin and surgical simulation . The silAfter the cleaning of the two parts of the model, the inferior turbinates were filled up with Dragon Skin FX-Pro, to make them more compressible. The two parts were then glued together shaping the entire model. To replicate the nasal mucosa, some layers of silicone were added over the nasal septum. Lastly, the skin portion was placed on the lid and the box was closed .As an additional feature, we implemented an Augmented Reality (AR) view to be integrated into the simulator. This allows a \u201chybrid\u201d training, i.e. the fusion of digital and physical tools, that may improve the potentialities of the simulator.The anatomical virtual models and some cutting planes planned by the surgeon on the reconstructed anatomy  were impTo track the physical simulator, we designed and printed a tracker with a well-contrasted texture, which was then anchored to the external box of the simulator .The tracking algorithm and the registration between the digital models and the real scene were implemented using the Image Target function of Vuforia engine: the engine detects the image texture of the tracker by comparing extracted natural features from the camera image against the known image target . Once the image target is detected, Vuforia engine will track the image target on the tracker and augment the scenario with digital content anchored to the target itself.The AR application was built as a UWP  app deployed on Microsoft HoloLens 2 smart glasses. The user just needs to look at the tracker on the simulator while wearing the AR glasses, until the image target matching occurs. After that, tracking of the real object and virtual content overlay can begin.The AR application generates holographic overlays on the physical model, by rendering the underlying anatomical structures , not visible from the outside of the simulator, together with the cutting planes which may help the trainee in understanding the deep anatomical structures while navigating the physical model with instruments see .Interactable user interface toggles (check boxes) were added to turn off and on the rendering of each virtual anatomical structure and cutting planes. In particular, cutting planes following progressive lines perpendicular to the nasal bones were created to convey the anatomical configuration that the surgeon has to bear in mind during the endoscopic approaches to the frontal sinus . The possibility to virtually remove the middle turbinate allows for realistic visualization of the intraoperative field across the steps to performing DRAF procedures.Voice commands to show/hide the virtual structures were also implemented in order to provide a completely hands-free AR application. see .The process to assess the fidelity and utility of the simulator as a training tool for ESS consisted of two phases which are described below.First, the anatomical realism of the nasal cavity and the reproducibility of the instruments handling inside it were evaluated.1 Identification of normal anatomy landmarks on one nasal cavity ;2 Removal of a foreign body  from the choana;3 Medialization of the middle turbinate and insertion of a cottonoid into the middle meatus;4 Incision of the simulated nasal septum mucosa with a curved scalpel and its detachment from the underlying simulated cartilage with a Cottle dissector-elevator.This phase involved 12 participants with different expertise levels: 3 ENT residents from the third and fourth year of residency, 3 young ENT specialists (less than 3 years of practice), and 6 ENT surgeons experienced in ESS (more than 10\u00a0years of practice). Each participant was asked to perform the following non-destructive tasks on the simulator using the standard instruments , in a tiThe completion of each task (yes/no) and the time needed to complete it were annotated by an external observer not involved in this phase (N.E.). A 5-point Likert scale questionnaire evaluating the anatomical, haptic feedback and utility of the simulator for training was filled in by each participant immediately after performing the non-destructive tasks .This phase was aimed at assessing the feasibility of different ESS procedures on the simulator and its potential as a training tool for ESS. Only four expert surgeons participated in this second phase. They were asked to simulate all the basic procedures of ESS, namely uncinectomy, maxillary antrostomy, anterior and posterior ethmoidectomy, sphenoidotomy  and DRAFAfter completing these tasks, the experts were required to fill out the task-specific questionnaire , using at-test for unpaired data was used to compare mean times for each pair of participant groups  for each task.All measured times needed to complete the tasks were reported as mean values and standard deviation (SD). p-value of <0.05 was considered statistically significant.All statistical tests were performed using SPSS software, version 26.0 , and a All the 12 participants completed the face and anatomical content validation of the model and answered the survey. The non-destructive tasks were accomplished by 100% of participants . The surn = 11) of the participants agreed or strongly agreed about the high anatomical fidelity of the simulator and the tactile feedback of both the turbinates and bony structures. On the other hand, 25% (n = 3) of the participants were neutral about the tactile feedback of the nostrils and nose tip and the use of instruments and endoscope inside the cavities. All the participants gave positive feedback (100% agreed or strongly agreed) regarding the role of the simulator in learning anatomy and improving accuracy and precision in endoscopic settings. Moreover, the utility of the simulator in developing hand-eye coordination received the highest scores (100% strongly agreed) from all responders.A total of 92% , the training has been traditionally performed either on ovine models or cadaveric heads, which both lack from pathology and specific anatomical variants, and could be dissected just once. The development of synthetic models for ESS training has encountered several difficulties in relation to the replication of the complex anatomy and of the variable haptic feedback from different parts of the nose and paranasal sinuses.The first low-fidelity models date back to 10\u00a0years ago, and despite having been demonstrated to improve basic endoscopic skills, the lack of anatomical and haptic fidelity hampered the possibility to simulate ESS procedures . More reThe unmet needs of 3D printed simulators for ESS encouraged our group to develop a 3D printed multi-material and multi-color patient-specific simulator for ESS, using the Polyjet printing technology. The potentiality and the precision of this recent technology allowed us to create a model with bony and soft structures simultaneously, reaching a high level of anatomical fidelity and an overall satisfactory realistic haptic feedback, as suggested by our results. Indeed, the technology has many benefits, including excellent resolution (up to 0.016\u00a0mm), smooth surfaces, and the ability to print multi-material and multi-color parts, thanks to its multiple print heads. Furthermore, it is possible to dynamically mix materials and create \u201cdigital materials\u201d with new characteristics. Indeed, it is possible to print rubber-like materials targeting a specific Shore-A hardness, as well as to mix up rigid and rubber-like materials, creating new materials with hybrid properties.However, we decided not to use the rubber-like Agilus Stratasys material, which has a minimum achievable Shore-A 30, to reproduce the facial skin, since preliminary tests with surgeons confirmed that the obtained 3D printed part is still too rigid if compared to the real facial skin. Moreover, using Agilus material at the minimum Shore-A 30, the parts can be printed only in white, so it is not possible to achieve a realistic color for the skin. Many papers in the literature report the use of the Dragon Skin FX-Pro, especially for the replication of the human skin using specific pigments to have different realistic skin coloration .Regarding the turbinates, we decided to fill them with silicone because the filling with Agilus Shore-A 30 made them too rigid. Conversely, the silicone filling allows the turbinates to be more compressible even if they still maintain a greater elasticity if compared to the real ones.Moreover, the silicone has realistic feedback to cut, and we decided to use it also for simulating the nasal mucosa; also in this case, using an additional layer of Agilus Shore-A 30 on the nasal septum is not a good solution to achieve a realistic replication of the mucosa.In our simulator, we were able to replicate the mucosa on the nasal septum so that the simulation of mucosal incision and mucoperichondrial flap elevation could be successfully performed on the 3D printed model by all participants. To the best of our knowledge, this study is the first to consider the feasibility of endoscopic lacrimal surgery on a 3D printed model. The nasolacrimal duct and sac were realized with a different material (VeroYellow and Agilus30 with Shore-A 35), obtaining a color quite similar to the surrounding bone, as it is in real anatomy. The consistency of the sac resulted quite fragile so that in one case it was accidentally drilled before identifying it.in vivo and on cadaver).Moreover, in our model some high-risk anatomical landmarks such as the lamina papyracea and the dura mater of the anterior skull base were replicated with layers of material that, despite differences in color from the real structures, resulted not easily identifiable during endoscopic dissection, as in a real-life situation during ESS , and about 20\u00a0h for the design and printing of the external box and face mold.A strategy to reduce these times is to take advantage of the modular design of the simulator, in which the internal part relating to the patient-specific paranasal sinuses anatomy can be changed and customized from time to time, while the external part (box and face mold) can be reused.Despite all our efforts in the cleaning of the paranasal sinuses model, some residual printing support material remained inside some ethmoidal cells, reducing the anatomical reliability and the performance of selected ESS steps. Future studies will include improvements in the design of the model to entirely clean it and reproduce the variable degree of pneumatization of the paranasal sinuses.The performance of ESS procedures was evaluated by a subjective Likert scale applied to not-validated questionnaires. However, for the purposes of this preliminary study, these methods could be considered acceptable in identifying positive feedback from the users. This limitation is shared with several other validation studies on the same topic .Our results are promising but the evaluation was performed on a restricted number of participants.A future study will be planned to compare the ESS training based on our simulator and the one based on cadaver dissection, involving a higher number of trainees.The AR feature we introduced can certainly be improved, e.g. by implementing the possibility to manipulate the virtual objects in addition to making them visible or not. Although we have received enthusiastic feedback from surgeons who have also experimented with the AR feature, its educational and training benefits must be more systematically evaluated.This study demonstrated that a detailed multi-material 3D printed model of sinus anatomy can be generated with a high level of anatomical accuracy and an overall satisfactory haptic response. This technology has the potential to be used in surgical training of ESS procedures, as an alternative or complementary training solution to cadaveric dissection. It could also potentially play a significant role in preoperative planning, since it may provide an individualized preoperative design on which the surgeon can rehearse before surgery, to shorten operation duration and ensure safe and effective surgery in patients."}
+{"text": "Among the various risk factors associated with contrast-induced acute kidney injury (CI-AKI), the importance of osmolality and viscosity is emerging among the characteristics of contrast media (CM) itself. High osmolality CM (HOCM) is deprecated and low osmotic pressure (LOCM) and iso-osmotic pressure (IOCM) are mainly used in clinical situations where the results of studies on their effect on the development of CI-AKI are contradictory. We evaluated the association between the type of CM and the risk of CI-AKI.A retrospective observational cohort study to analyze the effect of the type of CM on the development of CI-AKI. Using propensity score (PS) matching, 2,263 LOCM and IOCM groups were paired for analysis from 5,267 patients and fulfilled the inclusion criteria among 12,742 patients who underwent CAG between 1 January 2007, and 31 December 2016. LOCM included iopromide and iopamidol, IOCM was iodixanol. CI-AKI, which was the primary endpoint, was defined based on the Kidney Disease Improving Global Outcomes criteria within 48 h after exposure to the CM. A multivariable logistic regression analysis was used in the unmatched and matched cohorts, respectively. In addition, a stratified model on clinically important variables, including a high Mehran score (\u2265 6), was also used in the matched cohort.p = 0.006), but it lost statistical significance after PS matching . In multivariable analyses, the adjusted odds ratio for CI-AKI in the LOCM group were 1.059  in unmatched cohort and 0.987  in matched cohort. These results were also consistent with the high-risk (high Mehran score) group.LOCM users showed an increased incidence of CI-AKI (11.7% vs. 9.3%; Although the role of CM types in the development of CI-AKI has been debated, our observation shows that the selection between LOCM and IOCM during CAG has no influence on the incidence of CI-AKI. Contrast-induced acute kidney injury (CI-AKI) is an acute impairment of renal function after administration of intravascular iodine contrast media (CM) without other causes. CI-AKI is the third most common cause of in-hospital AKI , and hasThe risk of CI-AKI is affected by the patient- and procedure-related factors and the most important factor of CI-AKI is pre-existing renal impairment , 5. Amonn = 6,141), (2) patients exposed to CM within 7 days before or 3 days after the procedure (n = 963), (3) patients with receiving dialysis before study entry (n = 301), (4) patients without data for the type of CM (n = 70). Therefore, 5,267 patients were included in the final analysis (We conducted a retrospective propensity score (PS)-matched study at a single center to analyze the difference between LOCM and IOCM in the development of CI-AKI. All patients who underwent coronary angiography (CAG) or percutaneous coronary intervention (PCI) between 1 January 2007, and 31 December 2016, were screened for eligibility for the study. The total number of screened patients was 12,742 and 7,475 patients had been excluded based on the following exclusion criteria: (1) patients without pre- and post-procedural laboratory findings . Because this study did not involve any further intervention in the retrospective analysis, the need for obtaining consent was waived by the IRB of Gachon University Gil Medical Center. All methods were performed as per relevant guidelines and regulations.The patients' demographic, clinical, and laboratory data were obtained from electronic medical records. Demographic data included age, sex, body mass index (BMI), smoking status, comorbidities , and preprocedural prescript medication [renin-angiotensin-aldosterone system (RAAS) blockers [angiotensin-converting enzyme inhibitors (ACEi) or angiotensin receptor blockers (ARB)], beta-blockers, calcium channel blockers (CCB), diuretics, and statins]. We also collected the patients' clinical information about left ventricular ejection fraction (LVEF), left ventricular mass index (LVMI), blood pressure (BP) at the time of CAG, presence of multivessel disease, and the type and amount of contrast media used. Hypertension was defined as a documented blood pressure \u2265 140/90 mmHg or using anti-hypertensive medications. History of CHF or LVEF \u226440% on echocardiography was considered as heart failure.2, proteinuria (defined as trace or greater by dipstick), or both on at least 2 occasions \u2265 3 months apart.Laboratory data included serum creatinine levels before and after CAG or PCI, as well as hemoglobin, albumin, cholesterol, triglyceride, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol levels at hospital admission. We calculated the estimated glomerular filtration rate (eGFR) using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. Chronic kidney disease (CKD) was defined as either eGFR \u226460 ml/min/1.73 mThe Mehran CI-AKI risk scores were calculated for each patient from the corresponding scores for the 8 prognostic variables suggested in the previous report .Each patient followed the principle of hydration with isotonic saline for 12 h before and after exposure to CM, and the amount and rate were adjusted by the clinician according to the patient's tolerance. Each patient underwent CAG from the arterial access of either the femoral or radial arteries. The number of diseased coronary arteries was categorized as per the American Heart Association classifications. For significant stenotic coronary lesions, PCI was performed with a balloon catheter or stent.The non-ionic monomeric LOCM, namely iopromide (774 mOsm/kg) or iopamidol (800 mOsm/kg), was used in 54.2% of the total patients. The others have used the non-ionic dimeric IOCM, iodixanol (290 mOsm/kg) for coronary catheterization.The primary endpoint was CI-AKI defined as Kidney Disease Improving Global Outcome (KDIGO) criteria, an absolute increase of serum creatinine level \u2265 0.3 mg/dl or relative increase \u2265 50% from baseline creatinine level within 48 h after exposure to the CM.t-test, and McNemar's tests for continuous variables and categorical variables, respectively.Because the study was a retrospective observational study, we used the PS matching analysis to control any potential confounding and selection bias. A multivariable logistic regression analysis model was generated to predict the probability that LOCM would be administrated for the given covariates: age; sex; BMI; diabetes; hypertension; smoking status; CHF; baseline levels of eGFR, hemoglobin, albumin, cholesterol, triglycerides, HDL cholesterol, and LDL cholesterol; use of RAAS blockers, beta-blockers, CCBs, diuretics and statins; LVEF; LVMI; blood pressure; amount of CM used; presence of multivessel disease; PCI; and Mehran score. According to these covariates, a PS was calculated for each patient. And we used the derived PS values to match 2,263 IOCM users with LOCM users at a ratio of 1:1 using the nearest neighbor with calipers method . After all PS matches were performed, we conducted the balance test in baseline covariates using the standardized mean difference, paired t-test and categorical variables were compared by the \u03c72 test. To determine independent risk factors for CI-AKI, a multivariable logistic regression analysis was used in the unmatched cohort and matched cohort, respectively. A logistic regression model stratified on clinically important variables, including a high Mehran score (\u22656), was also used in the matched cohort. All statistical analyses were performed using R software, version 4.1.0 with packages (The Comprehensive R Archive Network: http://cran.r-project.org). Statistical significance was defined as p < 0.05.All continuous and categorical variables were expressed as the mean \u00b1 standard deviation and absolute counts with percentages, respectively. Before PS matching, continuous variables were compared by The baseline characteristics of the study population are summarized in p = 0.006), but it lost statistical significance after PS matching  in matched cohort  and 0.877 , respectively  were diagnosed with CKD. A total of 950 of 1,905 (49.9%) PS-matched CKD patients received LOCM, while 1,313 of 2,621 (50.1%) PS-matched non-CKD patients received LOCM . The ORsAs sensitivity analyses, we also analyzed the high-risk group for CI-AKI (Mehran score \u22656), but there was no difference in the incidence of CI-AKI between the LOCM and the IOCM groups .p = 0.725). Furthermore, according to subgroup analyses, the types of administrated CM did not increase the occurrence of CI-AKI among patients who had risk factors for CI-AKI. These findings are expected to help provide useful information for selecting the types of CM in patients with CAG.In this study, we assessed the role of the types of CM in the development of CI-AKI among patients who underwent CAG or PCI. Using PS matching, we found that no significant difference existed in the occurrence of CI-AKI between LOCM and IOCM  or STEMI, may have an influence on the results. Second, patients in the study were included regardless of their risk factors in contrast that most randomized controlled trials included high-risk patients. Considering that the incidence of CI-AKI in patients with normal kidney function was under 2%, the effect of the types of CM might be likely to be underestimated. Third, pre-existing AKI may be included at the start of the observation because the occurrence of AKI was estimated based on serum creatinine at the time of admission for CAG. Despite these limitations, our study is the only large observational cohort study using PS matching that provides evidence that no significant difference existed in the occurrence of CI-AKI between LOCM and IOCM.In conclusion, although the role of CM types in the development of CI-AKI has been debated, our observation shows that the selection between LOCM and IOCM during CAG has no influence on the incidence of CI-AKI. These findings are expected to help provide useful information for selecting the types of CM in patients with CAG.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Institutional Review Board of Gachon University Gil Medical Center. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements. Written informed consent was not obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.JYJ conceived and designed the experiments. TL and JYJ contributed to data analysis/interpretation and manuscript writing. Each author provided important intellectual content by presenting and solving questions about the accuracy or integrity of all parts of the work. All authors contributed to data acquisition, reviewing of the draft, and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "This study simulated the storage conditions of chilled beef at retail or at home, and the sterilization and preservation effects of short-time ultraviolet irradiation were studied. The conditions of different irradiation distances  and irradiation times  of ultraviolet (UV) sterilization in chilled beef were optimized, so as to maximally reduce the initial bacterial count, but not affect the quality of the chilled beef. Then, the preservation effect on the chilled beef after the optimized UV sterilization treatment during 0 \u00b1 0.2 \u00b0C storage was investigated. The results showed that UV irradiation with parameters of 6 cm and 14 s formed the optimal UV sterilization conditions for the chilled beef, maximally reducing the number of microorganisms by 0.8 log CFU/g without affecting lipid oxidation or color change. The 6 cm and 14 s UV sterilization treatment of the chilled beef was able to reduce the initial microbial count, control the bacterial growth, and delay the increase in the TVB-N values during storage. Compared with the control group, the total bacterial count decreased by 0.56\u20131.51 log CFU/g and the TVB-N value decreased by 0.20\u20135.02 mg N/100 g in the UV-treated group. It was found that the TBARS value of the UV treatment group increased during late storage; on days 9\u201315 of storage, the TBARS values of the treatment group were 0.063\u20130.12 mg MDA/kg higher than those of the control group. However, UV treatment had no adverse impact on the pH, color, or sensory quality of chilled beef. These results prove that UV treatment can effectively reduce the microbial count on the surface of beef and improve its microbial safety, thus maintaining the quality of beef and prolonging its shelf life. This study could provide a theoretical basis for the preservation technology of chilled beef in small-space storage equipment. The characteristics of chilled beef, such as soft and elastic texture, low juice loss, high nutritional value, and delicious taste, make it popular among consumers ,2. HowevTraditionally, ultraviolet (UV) light irradiation has been used for water, air, and surface sterilization . UV ligh2 for 0, 5, 10, 15, and 20 min in their study, and the irradiation time was directly proportional to the inhibition effect. Yeh et al. [2 for 30 s can be used to improve Salmonella control in ground beef, with individual applications of UV light reducing approximately 1 log CFU/g. In another study, cut pieces of food samples contaminated with E. coli or S. aureus were submitted to UV-C (254 nm) irradiation for 0, 1, 2, 3, 4, 5, and 10 min, and the results showed that the number of E. coli in beef was reduced by (1.0 \u00b1 0.2) log10 CFU/mL after 5 min of exposure, and in chicken and pork, the number was reduced by (1.6 \u00b1 0.7) log10 CFU/mL and (1.6 \u00b1 0.4) log10 CFU/mL after 4 and 10 min of irradiation, respectively [2 dose) in reducing the microbial loads in raw and smoked salmon; L. monocytogenes were reduced by 0.2 to 1.1 log on raw salmon and 0.7 to 1.3 log on smoked salmon; and the shelf-life was increased by up to 7 days and 14 days for raw and smoked salmon, respectively.Compared with thermal treatment or the application of antimicrobial compounds, UV irradiation is a very simple and safe sterilization process due to its ready availability and non-generation of potentially hazardous chemical residues ,11. As ah et al.  reportedectively . Holck eectively  studied UV-C light inactivates microorganisms by damaging their nucleic acid, which absorbs UV light; nucleic acids are the strongest 253.7 nm light absorbers. A UV lamp emitting a wavelength of 254 nm is very close to the maximum absorption of nucleic acids; therefore, such UV lamps can be widely used in food sterilization and disinfection. Although several studies have employed UV irradiation to reduce bacteria on meat surfaces and improve microbial safety ,20,21,22Chilled beef longissimus dorsi was purchased from local supermarkets. The meat samples were trimmed of visible connective and adipose tissue, cut parallel to the muscle into equal portions of 250 g (approximately 8 cm \u00d7 3 cm \u00d7 3 cm), and individually packaged in fresh-keeping bags .A total of thirty samples were selected and randomly assigned to the control and nine UV treatment groups in the step of evaluation of UV sterilization conditions. A total of 42 samples were selected and randomly assigned to the control and treatment groups in the step of evaluation of preservation effect, the control group and the treatment group consisted of 21 beef samples each. Samples were stored in the refrigerator before use, and the temperature was 0 \u00b0C (\u00b10.2 \u00b0C).The storage condition of meat sell at retail or storage at home were simulated in the study, and the research was carried out in a small space (in a refrigerator), so that 6\u201312 cm irradiation distances were chosen.Nine groups of UV sterilization conditions were set up according to different irradiation distances  and irradiation times , and the samples packaged in fresh-keeping bags without UV sterilization were used as the control group. UV treatment was carried out using the device shown in Chilled beef packaged in fresh-keeping bags was treated with the optimal UV sterilization parameters (UV at 6 cm for 14 s); the same batch of chilled beef without UV treatment was used as a control. The samples were stored at 0 \u00b0C (\u00b10.2 \u00b0C) in a refrigerator; then, the total bacterial count, TVB-N value, TBARS value, color, pH value, and sensory evaluation were measured at 0, 3, 6, 9, 11, 13, and 15 days of storage.The parameters of the UV lamps were set as 254 nm wavelength, 150 mm length, and 5 W total power. The setting positions of the five UV intensity test points were shown in Test point a is the left endpoint of the meat; test point b is the center point of the meat; test point c is the right endpoint of the meat; test point d is the endpoint of the meat near the inside of the box; and test point e is the endpoint of the meat near the outside of the box.The total bacterial count in chilled beef was determined according to the China National Food Safety Standard method GB 4789.2-2016 Food Microbiological Inspection Determination of aerobic plate count. Twenty-five grams of each sample was weighed and transferred to a homogeneous bag containing 225 mL of normal saline under aseptic conditions. After serial 10-fold dilution, 1 mL of the suspension was added to the plate, and PCA medium was poured in. After it was mixed and solidified, the number of bacteria was counted after incubation at 37 \u00b0C for 48 h. The results were expressed as the logarithmic number of colony forming units (CFUs).The standard for sensory evaluation of the freshness of the chilled beef was established according to the China National Food Safety Standard method GB 2707-2016 Fresh (Frozen) Livestock and Poultry Products. A 10-point scale  was used0*, \u2206a = a* \u2212 a0*, \u2206b = b* \u2212 b0*. L*, a*, and b* are the measured values of the sample; L0 *, a0 * and b0 * are the measured values of the control group.Color parameters were measured using a colorimeter . The aperture opening size was 8 mm, and the light source was D65. The observer angle used was perpendicular to the surface of the samples to obtain an accurate recording of the values. Five repetitions were performed for each sample, and color changes were described by L*, a*, and b*. The difference in color (\u2206E) between the UV-treated and control samples was calculated by the following equation:X is the content of malondialdehyde in the sample (mg MDA/kg), c is the concentration of malondialdehyde in the sample solution obtained from the standard series curve (\u00b5g/mL), V is the volume of the sample solution (mL), and m is the mass of sample in the final sample solution (g).The TBARS determination of the chilled beef followed the procedure used by Erkan et al.  with somn = 3) were expressed as mg of N per 100 g of sample.Total volatile basic nitrogen (TVB-N) was measured according to the China National Food Safety Standard method GB 5009.228-2016 Determination of total volatile basic nitrogen in food. Five grams of the minced sample was dispersed in 50 mL distilled water for 30 min, then 10 mL of filtered supernatant was pipetted into the distillation tube. One gram of magnesium oxide was added to the distillation tube, and analysis was performed using a Kjeldahl automatic nitrogen analyzer . The results  directly into the meat.n = 3). The significance of the main effects was determined using the one-way ANOVA procedure, and the determination of significant differences (p < 0.05) among the means was performed by Duncan\u2019s multiple-range and Tukey test. The graphs were generated using GraphPad Prime 8.The results were statistically analyzed using SAS 8.0 software and expressed as mean \u00b1 standard deviation (p < 0.05). However, all test points obtained the lowest UV intensity when the distance between the UV lamp and the sample was 12 cm. When the irradiation distance was 6 cm, test point b received the largest area of exposure among all the test points, and the UV intensity detected at point b was the highest. Other points  received a smaller radiation area, so the measured data were smaller than point b. As the irradiation distance increased, the transmission of UV light decreased, and the measured differences gradually decreased.The measurement of the intensity of UV radiation was taken inside the PE bag. As shown in p < 0.05); the total bacterial count was reduced by approximately 0.8 log CFU/g. Sobeli et al. [2 on the aerobic mesophilic bacterial counts of beef loin steaks; the highest microbial inactivation of 3.49 \u00b1 0.67 log CFU/g was determined under pulsed UV-C treatment at 4.2 J/cm2. Wang et al. [2, UVC treatment (254 nm wavelength) reduced aerobic bacterial counts by 0.64 to 1.00 log CFU/cm2. Although the samples used in our study varied, the chilled beef samples purchased from a market were used instead of inoculated fresh beef tissues and achieved a similar sterilization effect at a lower UV exposure dose.i et al.  investigg et al.  concludeg et al.  reported2 of UV-C compared with the control (0.27 mg MDA/kg), so they pointed out that as the dose of UV-C increases, lipid oxidation gradually increases. In our study, under the same irradiation distance, the group with a longer treatment time showed higher mean TBARS values, while the group with a closer distance showed higher mean TBARS values within the same treatment time. However, the statistical results showed that no significant change was observed between each UV sterilization treatment group and the control group, indicating that UV treatment had no significant effect on the lipid oxidation of the packaged chilled beef. Lipid oxidation is a major cause of meat quality deterioration during processing and storage [The lipid oxidation of chilled beef in the different treatment groups is presented in  storage . Some st storage  and 2.5 The color parameters of the UV-C light treated and untreated samples are shown in The results of the total color difference showed that compared to the control group, the UV (12 cm and 14 s) treatment group obtained the smallest \u2206E; in addition, \u2206E in the UV (6 cm and 14 s), UV (9 cm and 6 s), UV (12 cm and 10 s), and UV (12 cm and 6 s) treatment groups were small.p < 0.05). There were no significant differences between the nine UV treatment groups and the control group for odor and texture.These results suggested that UV irradiation at a close distance (6 cm) and for a long period of time (14 s) was an effective method to reduce the initial bacterial count without affecting the lipid oxidation, color loss, or sensory evaluation of chilled beef packaged in a fresh-keeping bag. Therefore, the UV (6 cm and 14 s) treatment was selected to study the change in quality and freshness during storage in the following study.p < 0.05). The value increased to 6.14 log CFU/g at day 11, exceeding the acceptable threshold of 6.0 log CFU/g stated by the Chinese National Standard for spoilage. The total bacterial count in the control group significantly increased on day 6, as it reached 6.52 \u00b1 0.11 log CFU/g, while that in the treatment group was 5.13 \u00b1 0.43 log CFU/g. There were no significant differences in the total bacterial count between days 13 and 15 of storage, in spite of a slight reduction in the total bacterial count on day 15 of storage in the treated and control groups.The initial bacterial counts in the treatment and control groups were 4.73 \u00b1 0.38 log CFU/g and 5.29 \u00b1 0.16 log CFU/g, respectively. The changes in the total bacterial count of chilled beef during storage are shown in p < 0.05), suggesting a slower growth rate than the control group.During days 0\u201311 of storage, the total bacterial count of chilled beef sterilized with UV-C was 0.56\u20131.51 log CFU/g lower than that of the control group. From days 6 to 9 of storage, the difference between the treated and control groups regarding the total bacterial count was more than 1 log CFU/g  is generally considered the principal indicator for assessing the freshness and shelf life of meat . As showp < 0.05). The TVB-N value of the UV treatment group was 12.08 \u00b1 1.14 mg N/100 g at the end of the storage. The control group exceeded the acceptable limit of 15.0 mg N/100 g stated by China National Food Safety Standard GB 2707-2016 for fresh meat on day 13, suggesting rapid spoilage activities in the control group. TVB-N is produced by the degradation of proteins and other nitrogenous compounds caused by enzymatic degradation and microbial action in beef [The TVB-N value of the UV treatment group was lower than that of the control group during storage, significantly from days 9 to 15 ( in beef . During 2) and stored at 4 \u00b0C for 9 days, and reported that the UV-C-treated samples exhibited slight pH value variations during refrigerated storage. These studies supported the conclusion that UV-C light did not affect the important meat quality parameter of pH.As shown in 2 on pH in Nile tilapia fillets during 11 days of storage at 4 \u00b1 1 \u00b0C. UV-C-treated groups demonstrated greater (p < 0.05) pH values than the control until the third day of storage, and lower pH values (p < 0.05) after the third day. They pointed out that it was due to the fact that UV-C light can promote protein degradation, which increases the amount of free amino acids, leading to a higher initial pH in the UV-C-treated samples. In our study, it was also found that the treatment group had a higher pH value than the control group during 3 days; however, there were no significant differences found by the statistical analysis.Moreover, some studies have shown that UV treatment can lead to changes in pH. Soro et al.  investigp < 0.05). This result indicated that the rate of lipid oxidation of chilled beef treated with UV increased in the late storage stage. Lipid oxidation products, such as aldehydes, have a negative effect on the odor and sensory shelf life of beef [As shown in  of beef . However2) before storage at 4 \u00b1 1 \u00b0C for 6 d; there were no significant differences in the TBARS values among all groups. L\u00e1zaro et al. [Chun et al.  measuredo et al.  considero et al.  also didp < 0.05) and changed insignificantly until day 13. In the early stage of storage, compared to the control group, the samples of the UV-treated group had significantly higher a* and lower L* values (p < 0.05), possibly because UV treatment changed the oxidation of myoglobin and the activity of microorganisms in meat. On days 9, 13, and 15 of storage, the a* value of the UV-C treatment group decreased significantly (p < 0.05), at which time the TBARS also showed a significant increase  on the sensory evaluation of Korean native cattle beef. The odor did not change in the UV-treated and control groups during 4 days of storage at 4 \u00b0C, and decreased singnificantly until day 9. Within 4 days of storage, the control group showed greater tenderness decreases and succulence loss compared to the treated groups. Liu et al. [Sensory evaluation is the most direct way to evaluate the quality of chilled meat. u et al.  used pulIn this study, UV parameters of 6 cm and 14 s were selected as the optimal conditions for UV sterilization of chilled beef packaged in fresh-keeping bags. The results of the UV sterilization treatment before storage showed an inhibition of the total bacterial count and TVB-N content, and no adverse effects on the quality indicators of the chilled beef  during storage. Additionally, although fat oxidation accelerated during late storage, the TBARS value did not exceed 0.5 mg MDA/kg throughout storage. Hence, UV technology can effectively decrease the surface contamination of chilled beef, improve its safety, and extend its shelf life, reducing economic losses due to microbial contamination and the spoilage of meat. This study could provide a theoretical basis for the preservation technology of chilled beef, especially in refrigerators or other small-space storage equipment."}
+{"text": "The use of health surveys has been key in the scientific community to promptly communicate results about the health impact of COVID-19. But what information was collected, where, when and how, and who was the study population?To describe the methodological characteristics used in large health surveys conducted in Spain early on in the COVID-19 pandemic.Scoping review. Inclusion criteria: observational studies published between January 2020 and December 2021, with sample sizes of over 2,000 persons resident in Spain. Databases consulted: PubMed, CINAHL, Literatura Latinoamericana y del Caribe en CC de la Salud, Scopus, PsycINFO, Embase, Sociological Abstracts, Dialnet and Web of Science Core Collection. We analyzed the characteristics of the literature references, methodologies and information gathered in the surveys selected. Fifty five studies were included.Sixty percentage of the studies included had mental health as their main topic and 75% were conducted on the general adult population. Thirteen percentage had a longitudinal design, 93% used the internet to gather information and the same percentage used non-probability sampling. Thirty percentage made some type of sampling correction to reduce coverage or non-response biases, but not selection biases. Sixty seven percentage did not state the availability of their data.Consistent with the extensive use of non-probability sampling without any bias correction in the extraordinary setting created by COVID-19, quality population frameworks are required so that probability and representative samples can be extracted quickly to promptly address other health crises, as well as to reduce potential coverage, non-response and particularly selection biases by utilizing reweighting techniques. The low data accessibility despite the huge opportunity that COVID-19 provided for Open Science-based research is striking. Health surveys are a fundamental support tool for decision-making in health planning. They provide information on magnitude, distribution and trends in health, the social factors that determine them and the use of social services from the population's perspective. They permit identification of the main challenges for prioritizing activity, designing and developing intervention strategies, evaluating and allocating resources, and the main risk groups in terms of health, lifestyles, and access to health services .The highly significant role of surveys for Public Health was greater still with the COVID-19 pandemic due to the urgent requirement for its health impact outcomes to be conveyed . This coIn this respect, non-probability surveys conducted with volunteers via the internet proliferated: for example, via websites, mobile apps, and publicity on social media. These types of survey enable statistics to be accessed more rapidly and at the same time provide an inexpensive means of compiling data, although they are subject to selection and coverage biases. This does not happen with probability surveys, often used by health statistics services such as Gold Standard, since they enable valid inferences to be made about the population without having to include hypotheses in models , 11. FurPrior statistical reweighting is therefore necessary in non-probability sampling in order to obtain valid and precise estimates that eliminate, or at last reduce, these biases , 13. In Finally, the use of health surveys has been key in the scientific community to promptly communicate results about the health impact of COVID-19. But what information was collected, where, when and how, and who was the study population? This research question justified the study objective of this work as the performance of a scoping review to describe the methodological characteristics of large health surveys conducted in Spain at the beginning of the COVID-19 pandemic.We performed a scoping review  using thThe following databases were consulted: PubMed, CINAHL (Ebscohost), Literatura Latinoamericana y del Caribe en CC de la Salud (LILACS), Scopus, PsycINFO (Proquest), Embase (Elsevier), Sociological Abstracts (Proquest), Dialnet and Web of Science Core Collection. We selected biomedical and multidisciplinary databases because most of the surveys during the pandemic were related to social services and according to the following criteria:Databases with large coverage and large numbers of journals included: Pubmed, Scopus, Embase.Databases with Spanish journals and articles in Spanish included: LILACS, Scopus, Dialnet.Databases specializing in health literature: Pubmed, CINAHL, Embase.Databases specializing in socio-sanitary literature: PsyINFO, Sociological Abstract, WOS, Scopus.https://supertracker.spi.ox.ac.uk/surveys/) and open searches in Google. The searches were developed between January 2020 and December 2021. These coverage dates of the databases were given by the novelty of the subject, COVID-19. There were no language restrictions. The search strategy was conducted through a combination of controlled terminology (MeSH/Emtree) and free language representative of the concepts COVID-19, surveys, and Spain, and was adapted to the different databases consulted (This search was complemented with gray literature information sources: OpenGray (unpublished literature), Gray Literature Report, the University of Oxford Global Directory for COVID surveys  using a standardized, predefined form that included variables relating to characteristics:Literature references: link to publication, first author institution of work, date of publication, language, name of journal, type of publication , open access (yes/no), impact factor and position (highest quartile) .Survey: geographical area, study population, study design, sampling design, effective sampling size, sample weighting and other corrections, survey type, date information collected, response rate, waves or measurements, analyses performed, availability of microdata , 2.Information collected: objective of study, primary topic [defined as mental health , lifestyThe variables of the second paragraph (survey characteristics) were selected from the STROBE  list, giA total of 3,095 articles were identified following the search strategy described above. Two thousand nine hundred twenty-four articles were identified using scientific literature databases and 171 using gray literature. A full-text check was performed on 225 of them, i.e., 6.4 and 21.6%, respectively, for scientific literature databases and gray literature. Finally, 55 references were included for the analysis .Almost half of the surveys selected published their results in 2020 (45%) and all of them began field work in 2020, one third of them in March (32.7%), 78.2% during the lockdown (March to April 2020) and 90.9% during the first state of emergency (March to June 2020). In addition, 80% of surveys collected information on one occasion or through one measurement. The YouGov bi-weekly information study  was founAs regards the study population of the 55 surveys selected for the analysis, 74.5% of them addressed the general adult population as their study population, while 9.1% considered the healthcare professionals , 68, 69.The main topics among the selected surveys were mental health (60.0%), lifestyle habits (10.9%), wellbeing (7.3%), and quality of life, life satisfaction, perceived risk of infection, resilience and working conditions (3.6%). Information regarding the objectives, information blocks and scales or composite variables was also gathered and is available in As regards sampling design, four of the fifty-five surveys selected (7.3%) had a probability design , 46, 71 In respect of sampling size, 35 surveys had between 2,000 and 5,000 participants (effective sample), two being found with over 50,000 participants , 62, botAs regards the statistical analysis conducted, thirty-six surveys developed a multivariate model, the most frequent being binary logistic , linear The distribution of groups of observations in the health surveys usually differs from the distribution in the survey population due to several reasons . Weighting is one of the best ways to reduce variances and to correct for frame deficiencies. In that sense, 30% implemented some type of sampling adjustment . The mosLastly, most of the surveys included (67.2%) did not report on the availability of microdata.As far as we are aware, this is the first scoping review on health surveys relating to COVID-19 and their main methodological characteristics; actually, we found only one similar study dating from 2013 , albeit Our main purpose with this review was to describe the methodological characteristics of surveys conducted early on in the pandemic, hence the search was focused on 2020 and 2021, noting that all surveys started during the first year of COVID-19. In fact, four out of every five surveys (78.2%) were conducted during the 2 months of the first lockdown period (March and April 2020). This demonstrates the rapid response by and considerable effort that the scientific community invested in attempting to provide information about the impact of COVID-19 on the population's health, with particular emphasis on mental health evidenced by the fact that more than half of the surveys (60%) focused on this as their main topic. This response was possible thanks to the internet: nine out of every ten surveys (92.7%) used social media, media sampling to recruit participants, or online subscription panels via this channel. The use of these types of survey expanded to such an extent during COVID-19 lockdowns that, along with more social considerations such as increasingly widespread internet access and use, they took over from traditional survey methods. In this regard, our study found that only four of the fifty-five surveys reviewed were conducted over the phone (7.3%) and, as was to be expected, no face-to-face surveys were identified.However, despite the efforts made by official statistical institutions, for example the European Statistical System through its Quality Assurance Framework , the sciNon-probability surveys present two advantages: they can collect large samples and they can do this in a short period of time. This is evidenced in our review, which shows that one out of every five surveys (21.8%) had a sample size of over 10,000 people, bearing in mind that one of the inclusion criteria was having an effective sample size of over 2,000. By contrast, the main drawback of non-probability surveys is that they present significant issues in terms of selection and coverage biases, thus compromising the generalization of results to the study population . Our revDespite these limitations, non-probability sampling can complement probability sampling if it is designed as a means to offset known biases in probability sampling by focusing on survey participant profiles that tend to be under-represented in such surveys . This noAnother outcome of our review worth noting is the low proportion of longitudinal surveys identified (12.7%). Surveys repeated over time are more difficult to conduct and analyze, but they do permit evaluation of changes in study variables in the same population, a key area for being able to obtain an overview of the pandemic and not just of its characteristics at a given moment in time . A samplLastly, this review is in line with other studies that show the high volume of scientific output related to COVID-19 . In our p = q = 0.5, 95% confidence level , 0% sample loss because we refer to effective sample , and design effect two. If a lower sample size were required, it would be very easy to retrieve those studies through Rayyan and repeat the analysis. However, although our record facilitates identifying these studies through Rayyan, it is worth pointing out that barely one third of the surveys reviewed make their data openly available, and this hinders performing these studies or other analyses such as, for example, reweighting techniques which would provide more reliable estimates. This clearly reflects the ongoing lack of research based on Open Science . For example, we considered as the last exclusion criteria surveys with a sample size of < 2,000 individuals. Our objective was to select large health surveys in terms of guaranteeing that sampling errors in overall estimates were below three percentage points assuming  Science , despite Science .Conceptualization and funding acquisition: AC-L, MR, and CS-C. Methodology: AC-L, CS-C, AO-d-L-L, EM-R, and CH-C. Data curation and analysis: AC-L, CS-C, DY, AO-d-L-L, and EM-R. Writing\u2014original draft preparation: AC-L, CS-C, and DY. Writing\u2014review and editing: MR, AC-L, CS-C, AO-d-L-L, EM-R, and CH-C. Project administration: AC-L. All authors have read and agreed to the published version of the manuscript."}
+{"text": "Objective: Reverse oblique (RO) and transverse intertrochanteric fracture patterns constitute a challenge for the operating surgeon. Currently, no gold standard fixation method exists. This study aimed to retrospectively compare proximal femoral nail (PFN) to dynamic condylar screw (DCS) plating in the treatment of RO and transverse intertrochanteric fractures.Methods: A total of 61 patients fixated by PFN or DCS were included. Of these, 36 were treated with PFN , and 25 were treated with DCS . The mean follow-up time was 33.8 and 42.6 months (range: 24-108). Radiological evaluation included the quality of fracture reduction, neck-shaft angle change, posteromedial support presence, and bone union time. Complications such as mechanical failure, nonunion, and infection were noted.Results: The only significant differences between the fixation methods were the superiority of DCS over PFN in earlier fracture union time  and the superiority (p=0.007) of PFN in shorter hospital stay (3.4 days versus\u00a05.1 days). No significant difference was observed in radiological parameters. While similar mechanical complication rates were found, a significantly higher nonunion rate was detected with the DCS.Conclusion: The most crucial disadvantage of DCS was the high rate of nonunion. Closed fracture reduction in PFN seems to be the most critical parameter to prevent severe complications. The open reduction using DCS showed no advantages over closed reduction and PFN fixation in providing a more anatomical alignment in AO/Orthopaedic Trauma Association (OTA) 31-A3 fractures. However, we recommend PFN application in this type of fracture, since nonunion is more common in DCS. A reverse oblique (RO) or transverse intertrochanteric fracture is an unstable fracture pattern. According to the AO/Orthopaedic Trauma Association (OTA) classification, these fractures are labelled as AO/OTA 31-A3. A3 fractures are further subdivided into A3.1, A3.2, or A3.3, as to the presence of simple oblique, simple transverse, or multi-fragmentary/wedge fracture pattern. In A3 fractures, the lateral cortex integrity is compromised, unlike most stable intertrochanteric fractures of the same region. Among all hip fractures, RO trochanteric fractures have a frequency of less than 2%\u00a0. DynamicIn order to overcome this problem, a dynamic condylar screw (DCS) system with a 95\u00b0 lag screw, initially designed for distal femur fractures, has been frequently preferred for the treatment of AO/OTA 31-A3 fractures\u00a0. HoweverAlthough in vitro studies showing the biomechanical superiority of PFN in treating AO/OTA 31-A3 fractures are available in the literature\u00a0,11, no cThis study aimed to retrospectively review patients with AO/OTA 31-A3 fractures who underwent internal fixation with PFN or DCS and compare them in terms of radiological outcomes and complication rates. It was hypothesized that DCS would provide superior radiological outcomes but would lead to more mechanical and clinical complications than PFN. The rationale was that the DCS system, with its plate placed on the lateral wall of the femur, would provide more anatomical fracture reduction and better compression in the lateral cortex; however, the necessity of open surgical reduction would lead to various complications.Among the 101 AO/OTA 31-A3 fracture patients operated on between 2013 and 2018, only patients with a follow-up of more than two years and fixed with either the DCS system or the PFN system with two lag screws were included in the study. PFN fixation with open fracture reduction, concomitant lower extremity fracture, open fractures, pathological fractures, and PFN systems with a single lag screw were excluded. The study was conducted retrospectively on the remaining 61 patients.Among the 61 patients, 36 were treated with the PFN, and 25 were treated with the DCS system. The mean follow-up period was 33.8 and 42.6 months (range: 24-108). All operations were performed on a traction table with the patient in a supine position and using C-arm fluoroscopy. All surgeries were performed by three senior orthopedic surgeons related to trauma surgery. If closed fracture reduction had been achieved, internal fixation with an intramedullary implant PROFIN PFN nail  was inserted by a minimally invasive technique through the medial border of the greater trochanter  Figure .Postoperative treatment included early mobilization and low-molecular-weight heparin prophylaxis for deep vein thrombosis for two weeks. Weight-bearing was not allowed before postoperative week 4, and gradual weight-bearing was initiated according to radiographic signs of callus formation. Patients were followed up in the outpatient clinic periodically.Radiological evaluation was performed separately by two experienced orthopedic surgeons. These two surgeons performed independent radiological evaluations. They only came together at points where they thought differently and made a decision.On the first postoperative radiographic view, the quality of fracture reduction was graded as good, acceptable , or poor \u00a0, BesidesStatistical analysis was performed using two tests. Student\u2019s t-tests were used to compare the two groups with regard to mean age, mean follow-up duration, bone union time, and hospital stay. Fisher\u2019s exact tests were performed to compare the groups with regard to gender, the side of injury, the energy of the trauma, AO/OTA classification, lateral butterfly fragment, body mass index (BMI), fracture reduction quality, posteromedial cortical continuity, neck-shaft angle change, and complications. A difference was considered to be statistically significant when p<0.05.There was no significant difference between the groups in terms of mean age, gender distribution, affected side, and fracture type distribution.Mechanical complications were detected in five patients (14%) with the PFN system  and two plate breakage (8%) with the DCS system. The revision of the initial fixation was necessary for one patient with reverse Z-effect, one PFN lag screw cutout, and two DCS plate breakage. All revisions were done with a new DCS system. Three Z-effects and one reverse\u00a0Z-effect patients were followed until the bone union was achieved and prominent lag screws were removed; however, revision surgery was not necessary.Nonunion was only encountered with the DCS system in four patients 16%), which was significantly higher than the PFN system (p=0.02). One was revised with a PFN system, and the union was achieved; three were revised with hemiarthroplasty. Although the total number of patients who needed fixation revision was higher in the DCS system, there was no significant difference . All resolved with early surgical debridement and intravenous antibiotic treatment, without a need for implant removal  and a longer hospital stay were shown as disadvantages. This may be related to open reduction. Contrary to our hypothesis, there was no significant difference between these two fixation methods in terms of the reduction of quality.Mechanical complications such as varus malunion, lag screw cutout, and implant failure were found to be high , as A3 fThe inability to achieve anatomical reduction is one of the most critical risk factors that increase failure in RO fractures\u00a0,23. AccoThere were only two studies that clinically compared DCS to PFN in AO/OTA 31-A3 fractures\u00a0,12. In aAccording to the study by Elis et al., there was almost no difference between PFN and DCS regarding operation time, hospital stay, malunion, and neck-shaft angle measurements, similar to the current study . HoweverThis study had some limitations. First, the study was retrospective and thus susceptible to missing data and selection bias. Second, the sample size is relatively small due to the low incidence of A3 fractures\u00a0but one of the highest reported in the literature. Third, we did not have any data on surgical time, bleeding amount, and bone mineral density of the patients. Lastly, although open reduction was applied in patients who underwent DCS plating, anatomical reduction could not be achieved in all of them. This might be related to the difficulty of open reduction in these fractures as reported previously.The higher complication rate of nonunion seems to be the most crucial disadvantage of DCS plating in AO-OTA 31-A3 fractures, although a shorter union time was seen in patients with a union in this group. Open reduction with DCS plating provided no advantage in obtaining a more anatomical alignment. Mechanical failure rates were nonsignificantly different between DCS plating and PFN. However, we recommend PFN application in this type of fracture, since nonunion is more common in DCS."}
+{"text": "Adequate surgical view during various types of nasal procedures is essential for surgical operations to be performed in a safe, efficient, and successful manner. Minimizing bleeding during surgery is an important way of increasing visualization that is commonly achieved by using a vasoconstrictive agent to control intraoperative hemorrhage. Many otolaryngologists choose to employ topical cocaine to minimize bleeding during surgery owing to its vasoconstrictive properties, while simultaneously benefitting from its dual local anesthetic effects. The relative benefit of topical cocaine for otolaryngologic procedures when compared to other topical analgesics and vasoconstrictors remains a topic of discussion due to the multiple potential cardiac and central nervous system side effects associated with cocaine administration. Furthermore, there is not a scientifically backed maximal safe dose published; instead, most of the guidelines for intranasal cocaine use are based on untested clinical practice. Despite this, the short latency, adequate duration of action, and inherent vasoconstrictive and decongestive capabilities make cocaine a valuable anesthetic agent for use in clinical procedures. As the relative benefit of using topical cocaine compared to the use of other vasoconstrictors and analgesics for nasal procedures remains undetermined in the current literature, this leaves the need for a comprehensive review of research that explores the risks and benefits of using topical cocaine in nasal procedures based on clinical trials that compare intranasal cocaine with various other analgesics and vasoconstrictors. In 1884, the Austrian ophthalmologist Carl Koller described the tissue-numbing properties of cocaine by placing cocaine solution on the cornea and producing insensibility ; howeverIndicationsCocaine can be used for intranasal procedures to provide anesthesia to the surgical site while allowing vasoconstriction of nearby vessels. This reduces intraoperative bleeding and provides a clear view during endoscopic nasal procedures . CocaineAdministrationCocaine nasal solution ranges from 4% to 10%, with 4% being the most frequently utilized . Direct-Mechanism of actionThe chemical structure of cocaine is a tropane alkaloid with weak basic properties. In its free form, cocaine is insoluble in aqueous solutions, but when ionized with hydrochloride salt, it becomes soluble in aqueous solutions . CocaineAdverse effects and toxicityCocaine is a popular street drug related to its propensity to get one high with a feeling of extreme euphoria. This effect is achieved by inhibiting the reuptake of the catecholamine dopamine from the synaptic cleft between axons . This reAnother potential toxicity occurs through cocaine\u2019s ability to block sodium channels, thus classifying it as an antidysrhythmic drug . Its sloWhile central nervous system and cardiovascular toxicities represent the most common adverse effects of cocaine use and abuse, there have also been adverse effects reported in various other systems, including pulmonary, musculoskeletal, and renal systems. In the pulmonary system, pneumothoraxes are thought to occur due to patient\u00a0inhalation of cocaine; additionally, bronchoconstriction can occur post-administration, which can worsen asthma and asthmatic symptoms . MusculoContraindicationsBeta-blockers have been contraindicated in treating patients due to the potential adverse effects of vasoconstriction from cocaine combined with peripheral vasocontraction due to inhibiting the beta-adrenergic receptors. This would cause unopposed action of alpha-adrenergic mediated vasoconstriction, leading to increased hypertension and potential hypertensive crisis ,26. CocaClinical studiesThe efficacy of cocaine nasal solution in the reduction of pain and various adverse effects secondary to otolaryngologic procedures has been studied vastly. One study compared the efficacy of various formulations of topical cocaine solutions in reducing nasotracheal intubation-induced epistaxis . In thisAnother study compared the anesthetic and vasoconstrictive efficacy of intranasal cocaine with xylometazoline/lidocaine solution . This stThe effects of cocaine versus tetracaine/adrenaline on producing local anesthesia during septoplasty were explored in a 2006 study . A totalCocaine has many innate properties, making it a valuable option for otolaryngologic procedures. Among these properties is its ability to act as a local anesthetic, vasoconstrictor, and decongestant with a short latency and adequate duration of action at a relatively low cost. However, there are many adverse effects associated with cocaine use that influence the cardiac, central nervous, pulmonary, musculoskeletal, and renal systems. Multiple studies have been conducted to determine the risk/benefit of the use of cocaine with variable results. When comparing various cocaine solutions , it was shown that all concentrations were effective analgesics, with 4% holding preference due to its lower rate of hemodynamic and toxic adverse effects. However, when comparing cocaine to other analgesics such as xylometazoline/lidocaine, tetracaine, and co-phenylalanine, it was found that alternative analgesics take preference due to decreased risk of various cardiovascular adversities. Cocaine remains a viable option for otolaryngologists who wish to perform nasal procedures with localized anesthesia, but alternative choices may prove to be better options related to decreased risk profile and similar effects."}
+{"text": "Autoimmune enteropathy (AIE) defined by intractable diarrhoea and nonceliac enteropathy with villous atrophy, is a rare digestive disease. Case reports of this disease are sporadic and the clinical characteristics of AIE is seldom discussed.We evaluate the clinical, laboratory, histopathological features, response to therapy and outcome of AIE in children.We conducted a retrospective analysis of five children with AIE in our hospital. A comprehensive search of MEDLINE was performed using PubMed, through keywords of \u201cautoimmune enteropathy, pediatric or children\u201d. The clinical manifestations, endoscopic results, pathological results, and medication therapy of these children were collected and the cases were divided into two groups, infants (\u2264\u20091 year old) and children (>\u20091 year old).Five cases treated in our department: one case took eight years to make the final diagnosis; one case was positive for anti-intestinal epithelial cell (AE) antibody; three cases showed crypt apoptosis in histopathology; and two cases showed celiac-like changes. All cases were responsive to glucocorticoid therapy in the early stage of treatment, while three cases required immunosuppressant maintenance. After reviewing the literature, we performed a statistical analysis of 50 cases with a male-to-female ratio of 31:19. Among them, 35 patients (70%) were within 1 year of age, and their clinical manifestations were mainly watery stool , weight loss , abdominal distension , serum AE or anti-goblet cell (AG) antibody positivity , other immune-related antibodies , gene mutations , and family history . All the children showed different degrees of intestinal villous atrophy. Thirty-seven (74%) of the children were treated early, and their clinical symptoms were relieved. Comparing the cases between different age groups, it was found that the mortality rate of children with onset in infancy was higher (P\u2009<\u20090.05), and there was no difference in other autoimmune diseases, AE antibody positivity rates, and other antibodies between the two groups. In addition to survival rate between different age group (P\u2009=\u20090. 005), there was no difference in sex, autoantibody positivity rate, single gene mutation, or family history between the two groups (P\u2009>\u20090.05) through analysis of mortality and clinical remission cases.Endoscopic examination and mucosal pathological examination should be performed to diagnose AIE in children with watery stool and weight loss who fail to be treated with diet therapy. Immunotherapy is the core of medical management of AIE and can improve prognosis. Children with a poor prognosis in infancy should be actively treated to reduce mortality rates associated with AIE. Autoimmune enteropathy (AIE) is a rare intestinal disease characterised by intractable diarrhoea and immune-mediated intestinal mucosal damage . The incDiagnosis of AIE is challenging, requires small bowel biopsy, and often confused with other caoditions causing glycogenic diarrhea, celiac disease, inflammation bowel disease, gene mutations related immune deficiency disease, and so on.Given the rare nature of AIE, most articles on this disease have been isolated case reports or case series, lacking multi-centre statistical data, especially the clinical characteristics of children at different ages. Therefore, this article will further deepen our understanding of the disease by comparing the clinical characteristics of children of different ages at time of disease onset.Detailed clinical data of five children diagnosed with AIE in our hospital from January 2020 to August 2022 were collected. Articles from 1982 to December 2021 were searched on Pubmed with the keyword \u201cautoimmune enteropathy, Paediatric or children\u201d, and the case data of all children diagnosed with AIE were obtained. A total of 69 cases involving children have been reported \u201317. ExceA 2 -year-old girl developed gastroenteritis after an insanitary diet at the beginning of the disease and then developed chronic diarrhoea. The clinical manifestations were watery stools and abdominal distension, which could not be relieved by diet therapy, antibiotic application, or other treatments. The stool was yellow green watery with occasional mucus and no bloody substances. During repeated monitoring, no red or white blood cells were found in the routine examination of the stool. Physical examination on admission revealed severe malnutrition. Laboratory examination:serum AE antibody was positive, decreased serum albumin. Endoscopic examination showed that the duodenal villi were low and flat see Fig.\u00a0A, B. TheA 5-month-old girl presented with increasing frequency of defaecation along with slow weight gain. Feeding the patient a deeply hydrolyzed formula powder was not effective. Colonoscopy showed terminal ileal oedema, a pathology suggestive of active inflammation, and the symptoms were slightly relieved by dexamethasone(1\u00a0mg/kg/day). By the age of 9 months, after respiratory tract infection, the child had a large amount of watery stool and progressive weight decline. Physical examination at admission revealed severe malnutrition and a dehydrated appearance. Gastroscopy showed duodenal mucosal oedema, and pathological examination showed obvious atrophy of the duodenal mucosa, lymphocyte infiltration in the lamina propria, goblet cells, and crypt apoptosis.A 9 -month-old-boy, had intermittent watery stool with slow weight gain 2 months after birth. Repeated antidiarrheal and antibiotic treatments were not effective. No ulcer or erosion was found via gastroscopy or colonoscopy in the other hospitals. Physical examination on admission revealed severe malnutrition and mild dehydration. antibody and gene detection of celiac disease were negative. Gastroscopy showed low duodenal mucosa. Pathological examination showed celiac-like changes with few neutrophil infiltrations : two cases immunodysregulation polyendocrinopathy enteropathy X-linked (IPEX),three cases hypogammaglobulinemia, one case autoimmune polyendrocrinopathy-cadidiasis-ectodermal dystrophy (APECED), one case Kabuki syndrome, one case X-linked immunodeficiency, and one case hyper-IgE syndrome.42 cases received drug treatment after diagnosis Table\u00a0, 7 casesThe children in our study were divided into infant (\u2264\u20091 year old) and child (>\u20091 year old) groups. The chi-square test was used for the analysis Table\u00a0. The morAutoimmune enteropathy (AIE) is an autoimmune disease of unknown aetiology, characterised by villous atrophy of the small intestinal mucosal epithelium. Most of these cause chronic diarrhea and may be life-threating diarrhea in infancy.The main clinical manifestation of AIE is severe and long-term secretory diarrhoea, which is characterised by a large amount of watery stools, with a diarrhoea volume of up to 5,000 mL/day. The effects of fasting and water deprivation are not typically beneficial and the faecal aetiology is negative. Parenteral nutrition is required to maintain water and electrolyte balance. Intestinal malabsorption can result in low body weight and poor growth . In addiLaboratory tests for AIE lack specificity, and AE and/or AG antibodies are only present in the serum intestinal autoantibodies of some children. Positive serum AE or AG antibodies increase the possibility of AIE diagnosis, but is not necessary for diagnosis . It has AIE has no specific histological features; however, all patients have gastrointestinal mucosal injuries. Injury is usually confined to the mucosa, while deep ulcers or transmural inflammatory changes are rarely observed. The pathological diagnostic criteria of AIE in the past were blunt villi, lymphocytosis in the deep crypt, increased apoptosis of crypt cells, and increased lymphocytes in the surface intraepithelium; however, at present, the pathological tissues of AIE can be divided into four types: active duodenitis , celiac disease-like, graft-versus-host disease-like (obvious apoptosis can be seen in crypts), and mixed mode. At present, all four types have been reported in paediatric patients , 20, 28.These results suggest that the prognosis of patients with AIE depends on the severity of symptoms and signs of the digestive system, the severity and extent of histological lesions of the gastrointestinal tract, and the existence of parenteral involvement , 29. HowAt present, there is no consensus on the treatment of AIE, because children with AIE often suffer from stunting due to nutritional absorption disorder, and their developmental conditions such as weight and height are mostly lower than those of normal children of the same age. The use of immunosuppressants based on nutritional support is recommended , 23. At Overall, the clinical and histopathological manifestations of AIE in children, especially in infants, are quite varied. Early diagnosis and medication can reduce mortality and improve growth and development. Therefore, it is helpful for clinicians and pathologists to identify the clinical characteristics and different pathological patterns of AIE in its early stages.This paper has limitations. Since the clinical data of 45 children were reported in the previous literature, the clinical remission time and follow-up after drug addiction were not fixed, so the overall evaluation and long-term clinical remission rate of drug treatment were uncertain. Therefore, the follow-up will strengthen the follow-up work of the confirmed patients in our hospital and help to evaluate the medication and clinical prognosis."
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