bluuebunny/crossref_metadata_2025 huggingface dataset:
Vitamin A supplement after neonatal Streptococcus pneumoniae pneumonia inhibits the progression of experimental asthma by altering CD4+T cell subsets
Authors: Tian Yonglu , Tian Qinqin , Wu Yi , Peng Xin , Chen Yunxiu , Li Qinyuan , Zhang Guangli , Tian Xiaoyin , Ren Luo , Luo Zhengxiu
Published: March 2020

Studies demonstrated that pneumonia can decrease vitamin A productions and vitamin A reduction/deficiency may promote asthma development. Our previous study showed that neonatal Streptococcus pneumoniae (S. pneumoniae) infection promoted asthma development. Whether neonatal S. pneumoniae pneumonia induced asthma was associated with vitamin A levels remains unclear. The aim of this study was to investigate the effects of neonatal S. pneumoniae pneumonia on vitamin A expressions, to explore the effects of vitamin A supplement after neonatal S. pneumoniae pneumonia on adulthood asthma development. Non-lethal S. pneumoniae pneumonia was established by intranasal inoculation of neonatal (1-week-old) female BALB/c mice with D39. S. pneumoniae pneumonia mice were supplemented with or without all-trans retinoic acid 24 hours after infection. Vitamin A concentrations in lung, serum and liver were measured post pneumonia until early adulthood. Four weeks after pneumonia, mice were sensitized and challenged with OVA to induce allergic airway disease (AAD). Twenty-four hours after the final challenge, the lungs and bronchoalveolar lavage fluid (BALF) were collected to assess AAD. We stated that serum vitamin A levels in neonatal S. pneumoniae pneumonia mice were lower than 0.7µmol/L from day 2–7 post infection, while pulmonary vitamin A productions were significantly lower than those in the control mice from day 7–28 post infection. Vitamin A supplement after neonatal S. pneumoniae pneumonia significantly promoted Foxp3+Treg and Th1 productions, decreased Th2 and Th17 cells expressions, alleviated airway hyperresponsiveness (AHR) and inflammatory cells infiltration during AAD. Our data suggest that neonatal S. pneumoniae pneumonia induce serum vitamin A deficiency and long-time lung vitamin A reduction, vitamin A supplement after neonatal S. pneumoniae pneumonia inhibit the progression of asthma by altering CD4+T cell subsets.
Anthropogenic electromagnetic fields (EMF) influence the behaviour of bottom-dwelling marine species
Authors: Hutchison Zoë L. , Gill Andrew B. , Sigray Peter , He Haibo , King John W.
Published: March 2020

Many marine animals have evolved sensory abilities to use electric and magnetic cues in essential aspects of life history, such as to detect prey, predators and mates as well as to orientate and migrate. Potential disruption of vital cues by human activities must be understood in order to mitigate potential negative influences. Cable deployments in coastal waters are increasing worldwide, in capacity and number, owing to growing demands for electrical power and telecommunications. Increasingly, the local electromagnetic environment used by electro- and magneto-sensitive species will be altered. We quantified biologically relevant behavioural responses of the presumed, magneto-receptive American lobster and the electro-sensitive Little skate to electromagnetic field (EMF) emissions of a subsea high voltage direct current (HVDC) transmission cable for domestic electricity supply. We demonstrate a striking increase in exploratory/foraging behaviour in skates in response to EMF and a more subtle exploratory response in lobsters. In addition, by directly measuring both the magnetic and electric field components of the EMF emitted by HVDC cables we found that there were DC and unexpectedly AC components. Modelling, restricted to the DC component, showed good agreement with measured results. Our cross-disciplinary study highlights the need to integrate an understanding of the natural and anthropogenic EMF environment together with the responses of sensitive animals when planning future cable deployments and predicting their environmental effects.
Randomized Clinical Trial investigating Self-Assembling Peptide P11-4 for Treatment of Early Occlusal Caries
Authors: Doberdoli Dafina , Bommer Claudine , Begzati Agim , Haliti Fehim , Heinzel-Gutenbrunner Monika , Juric Hrvoje
Published: March 2020

Non-invasive caries treatment is a major focus in modern dentistry. The present study was designed to evaluate the effectiveness of monomeric self-assembling peptide P11-4 (SAP P11-4) in combination with fluoride varnish or polymeric self-assembling peptide matrix (SAPM) in treatment of non-cavitated occlusal caries. Ninety children and adolescents were included in this randomized, gold-standard-controlled clinical trial. Test Group 1 received SAP P11-4 and twice fluoride varnish at baseline and Day 180, Test Group 2 received SAP P11-4 on baseline and twice weekly SAPM (home-application), and Control Group received fluoride varnish on baseline and Day 180. Caries progression was measured by laser fluorescence, Nyvad Caries Activity, ICDAS-II-codes, and investigator assessments. Laser fluorescence changes demonstrated superior results for Test Group 1 and 2, as values decreased compared to an increase for the Control Group (p < 0.0005). ICDAS-II codes at Day 360 showed partial regression for Test Group 1 (6.7%) and Test Group 2 (20.0%) and partial progression for Control Group (23.3%) (p < 0.01). Nyvad Caries Activity yielded superior caries inactivation for Test Groups, compared to Control Group (p = 0.002). This trial showed that SAP P11-4, applied either in combination with fluoride varnish or twice weekly SAPM, was a superior treatment for early caries compared to fluoride varnish alone.
Author Correction: Modeling organic carbon loss from a rapidly eroding freshwater coastal wetland
Authors: Braun Katherine N. , Theuerkauf Ethan J. , Masterson Andrew L. , Curry B. Brandon , Horton Daniel E.
Published: March 2020

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Diatoms synthesize sterols by inclusion of animal and fungal genes in the plant pathway
Authors: Gallo Carmela , Landi Simone , d’Ippolito Giuliana , Nuzzo Genoveffa , Manzo Emiliano , Sardo Angela , Fontana Angelo
Published: March 2020

Diatoms are ubiquitous microalgae that have developed remarkable metabolic plasticity and gene diversification. Here we report the first elucidation of the complete biosynthesis of sterols in the lineage. The study has been carried out on the bloom-forming speciesSkeletonema marinoiandCyclotella crypticathat synthesise an ensemble of sterols with chemotypes of animals (cholesterol and desmosterol), plants (dihydrobrassicasterol and 24-methylene cholesterol), algae (fucosterol) and marine invertebrates (clionasterol). In both species, sterols derive from mevalonate through cyclization of squalene to cycloartenol by cycloartenol synthase. The pathway anticipates synthesis of cholesterol by enzymes of the phytosterol route in plants, as recently reported in Solanaceae. Major divergences stem from reduction of Δ24(28) and Δ24(25) double bonds which, in diatoms, are apparently dependent on sterol reductases of fungi, algae and animals. Phylogenetic comparison revealed a good level of similarity between the sterol biosynthetic genes ofS. marinoiandC. crypticawith those in the genomes of the other diatoms sequenced so far.
Bisphenol A induces coronary endothelial cell necroptosis by activating RIP3/CamKII dependent pathway
Authors: Reventun P. , Sanchez-Esteban S. , Cook A. , Cuadrado I. , Roza C. , Moreno-Gomez-Toledano R. , Muñoz C. , Zaragoza C. , Bosch R. J. , Saura M.
Published: March 2020

Epidemiological studies link long term exposure to xenoestrogen Bisphenol-A to adverse cardiovascular effects. Our previous results show that BPA induces hypertension by a mechanism involving CamKII activation and increased redox stress caused by eNOS uncoupling. Recently, CamKII sustained activation has been recognized as a central mediator of programmed cell death in cardiovascular diseases, including necroptosis. However, the role of necroptosis in cardiac response to BPA had not yet been explored. Mice exposed to BPA for 16 weeks showed altered heart function, electrical conduction, and increased blood pressure. Besides, a stress test showed ST-segment depression, indicative of cardiac ischemia. The hearts exhibited cardiac hypertrophy and reduced vascularization, interstitial edema, and large hemorrhagic foci accompanied by fibrinogen deposits. BPA initiated a cardiac inflammatory response, up-regulation of M1 macrophage polarization, and increased oxidative stress, coinciding with the increased expression of CamKII and the necroptotic effector RIP3. In addition, cell death was especially evident in coronary endothelial cells within hemorrhagic areas, and Evans blue extravasation indicated a vascular leak in response to Bisphenol-A. Consistent with the in vivo findings, BPA increased the necroptosis/apoptosis ratio, the expression of RIP3, and CamKII activation in endothelial cells. Necrostatin-1, an inhibitor of necroptosis, alleviated BPA induced cardiac dysfunction and prevented the inflammatory and hemorrhagic response in mice. Mechanistically, silencing of RIP3 reversed BPA-induced necroptosis and CamKII activation in endothelial cells, while inhibition of CamKII activation by KN-93 had no effect on RIP3 expression but decreased necroptotic cell death suggesting that BPA induced necroptosis is mediated by a RIP 3/CamKII dependent pathway. Our results reveal a novel pathogenic role of BPA on the coronary circulation. BPA induces endothelial cell necroptosis, promotes the weakening of coronary vascular wall, which caused internal ventricular hemorrhages, delaying the reparative process and ultimately leading to cardiac dysfunction.
The association between breastmilk oligosaccharides and faecal microbiota in healthy breastfed infants at two, six, and twelve weeks of age
Authors: Borewicz Klaudyna , Gu Fangjie , Saccenti Edoardo , Hechler Christine , Beijers Roseriet , de Weerth Carolina , van Leeuwen Sander S. , Schols Henk A. , Smidt Hauke
Published: March 2020

Several factors affect gut microbiota development in early life, among which breastfeeding plays a key role. We followed 24 mother-infant pairs to investigate the associations between concentrations of selected human milk oligosaccharides (HMOs) in breastmilk, infant faeces, and the faecal microbiota composition in healthy, breastfed infants at two, six and 12 weeks of age. Lactation duration had a significant effect on breastmilk HMO content, which decreased with time, except for 3-fucosyllactose (3FL) and Lacto-N-fucopentaose III (LNFP III). We confirmed that microbiota composition was strongly influenced by infant age and was associated with mode of delivery and breastmilk LNFP III concentration at two weeks, with infant sex, delivery mode, and concentrations of 3′sialyllactose (3′SL) in milk at six weeks, and infant sex and Lacto-N-hexaose (LNH) in milk at 12 weeks of age. Correlations between levels of individual breastmilk HMOs and relative abundance of OTUs found in infant faeces, including the most predominant Bifidobacterium OTUs, were weak and varied with age. The faecal concentration of HMOs decreased with age and were strongly and negatively correlated with relative abundance of OTUs within genera Bifidobacterium, Parabacteroides, Escherichia-Shigella, Bacteroides, Actinomyces, Veillonella, Lachnospiraceae Incertae Sedis, and Erysipelotrichaceae Incertae Sedis, indicating the likely importance of these taxa for HMO metabolism in vivo.
Estimation of the spawning time of Japanese eels in the open ocean
Authors: Higuchi Takatoshi , Yamada Yoshiaki , Watanabe Shun , Kojima Takahito , Tsukamoto Katsumi
Published: March 2020

To understand the spawning ecology of the Japanese eel, the spawning time of this species was estimated based on measurements of the ascending speed of eggs and previously obtained data. Two types of water temperature parameters were calculated assuming an arbitrary spawning time. The ‘incubation temperature’ of 53 eggs collected in the spawning area was estimated based on the developmental stage of each egg and experimentally determined relationships between water temperature and incubation duration. The ‘experienced temperature’ of eggs ascending in the water column after spawning was estimated based on an ascending egg speed of 3.69 m/h and spawning depth of 230 m determined from a pop-up satellite archival tag release experiment on silver eels conducted in the same area. The incubation and experienced temperatures of the eggs coincided only at 20:20–22:30 h, 3 days prior to the new moon. This period is only a few hours after the diel vertical migration of Japanese eels in the evening, when adults move up from a depth of ~800 m (approximately 5 °C) to shallower waters of 200–250 m depth (approximately 20 °C). Our findings will facilitate improvements in aquaculture techniques and the detection of eel spawning events in the open ocean.
Context-dependent selection as the keystone in the somatic evolution of cancer
Authors: Vibishan B. , Watve Milind G.
Published: March 2020

Somatic evolution of cancer involves a series of mutations, and attendant changes, in one or more clones of cells. A “bad luck” type model assumes chance accumulation of mutations. The clonal expansion model assumes, on the other hand, that any mutation leading to partial loss of regulation of cell proliferation will give a selective advantage to the mutant. However, a number of experiments show that an intermediate pre-cancer mutant has only a conditional selective advantage. Given that tissue microenvironmental conditions differ across individuals, this selective advantage to a mutant could be widely distributed over the population. We evaluate three models, namely “bad luck”, context-independent, and context-dependent selection, in a comparative framework, on their ability to predict patterns in total incidence, age-specific incidence, stem cell number-incidence relationship and other known phenomena associated with cancers. Results show that among the factors considered in the model, context dependence is necessary and sufficient to explain observed epidemiological patterns, and that cancer evolution is largely selection-limited, rather than mutation-limited. A wide range of physiological, genetic and behavioural factors influence the tissue micro-environment, and could therefore be the source of this context dependence in somatic evolution of cancer. The identification and targeting of these micro-environmental factors that influence the dynamics of selection offer new possibilities for cancer prevention.
Oxygen consumption rate of Caenorhabditis elegans as a high-throughput endpoint of toxicity testing using the Seahorse XFe96 Extracellular Flux Analyzer
Authors: Preez G. Du , Fourie H. , Daneel M. , Miller H. , Höss S. , Ricci C. , Engelbrecht G. , Zouhar M. , Wepener V.
Published: March 2020

Caenorhabditis elegans presents functioning, biologically relevant phenotypes and is frequently used as a bioindicator of toxicity. However, most C. elegans in vivo effect-assessment methods are laborious and time consuming. Therefore, we developed a novel method to measure the oxygen consumption rate of C. elegans as a sublethal endpoint of toxicity. This protocol was tested by exposing 50 larval stage one C. elegans individuals for 48 h (at 20 °C) to different concentrations of two toxicants i.e. benzylcetyldimethylammonium chloride (BAC-C16) and cadmium (Cd). Following exposures, the oxygen consumption rate of the C. elegans individuals were measured using the high-throughput functionality of the Seahorse XFe96 Extracellular Flux Analyzer. Dose-response curves for BAC-C16 (R2 = 0.93; P = 0.001) and Cd (R2 = 0.98; P = 0.001) were created. Furthermore, a strong, positive correlation was evidenced between C. elegans oxygen consumption rate and a commonly used, ecologically relevant endpoint of toxicity (growth inhibition) for BAC-C16 (R2 = 0.93; P = 0.0001) and Cd (R2 = 0.91; P = 0.0001). The data presented in this study show that C. elegans oxygen consumption rate can be used as a promising functional measurement of toxicity.
Relationship between Sleep Duration, Sun Exposure, and Serum 25-Hydroxyvitamin D Status: A Cross-sectional Study
Authors: Choi Ji Ho , Lee Bora , Lee Jae Yong , Kim Chang-Hoon , Park Bumhee , Kim Dong Young , Kim Hyun Jun , Park Do-Yang
Published: March 2020

Normal-range sleep duration is an important factor for general health and metabolism, and insufficient or excessive sleep is associated with chronic metabolic disease. Among the many factors that affect sleep duration, sun exposure plays an important role in maintaining regular circadian rhythm and is also involved in the production and activation of 25-hydroxyvitamin D [25(OH)D], which regulates various functions in the body. However, 25(OH)D is available through food and various nutritional supplements without sun exposure, so it is important to find out the complex relationship among sun exposure, vitamin D status, and sleep duration. The relationship between sun exposure, vitamin D status, and sleep duration was analyzed in the nationwide survey and examination of 25,534 study populations, after adjusting for demographic characteristics, physical characteristics, lifestyle status, and socio-demographic variables. Vitamin D status alone did not show the relationship with sleep duration, although there were statistical relationships in the various factors including sun exposure with sleep duration. There was a statistical difference in 25(OH)D according to sleep duration, only in low sun exposure group. Subjects with low sun exposure and excessive sleep duration comparatively lower 25(OH)D than those with normal-range sleep, even after adjustment for potentially confounding factors. Individuals with limited exposure to sunlight should maintain adequate vitamin D status to have an appropriate sleep duration for health.
Two-step extinction of Late Cretaceous marine vertebrates in northern Gulf of Mexico prolonged biodiversity loss prior to the Chicxulub impact
Authors: Ikejiri Takehito , Lu YueHan , Zhang Bo
Published: March 2020

Recent studies on mass extinctions are often based on the global fossil record, but data from selected paleogeographic regions under a relatively constant paleoenvironmental setting can also provide important information. Eighty-nine marine vertebrate species, including cartilaginous and bony fish and marine reptiles, from northern Gulf of Mexico – located about 500 km from the Chicxulub crater – offer a unique opportunity to determine an extinction process during the last 20 million years of the Late Cretaceous. Our diversity data show two separate extinction events: (i) the ‘Middle Campanian Crisis’ (about 77 Mya) and (ii) the end-Maastrichtian (66 Mya) events. Whether this stepwise pattern of extinctions occurred locally or globally cannot be determined at present due to the lack of a dataset of the marine vertebrate record for reliable comparison. However, this stepwise pattern including the Middle Campanian and end-Maastrichtian events for, at least, a 13 million-year interval indicates long-term global marine environmental changes (e.g., regression, ocean water chemistry change). Because most Cretaceous marine vertebrates already disappeared in the Gulf of Mexico prior to the latest Maastrichtian, the Chicxulub Impact may not be considered as the most devastating extinction event for the community.
Calprotectin, a new biomarker for diagnosis of acute respiratory infections
Authors: Havelka Aleksandra , Sejersen Kristina , Venge Per , Pauksens Karlis , Larsson Anders
Published: March 2020

Respiratory tract infections require early diagnosis and adequate treatment. With the antibiotic overuse and increment in antibiotic resistance there is an increased need to accurately distinguish between bacterial and viral infections. We investigated the diagnostic performance of calprotectin in respiratory tract infections and compared it with the performance of heparin binding protein (HBP) and procalcitonin (PCT). Biomarkers were analyzed in patients with viral respiratory infections and patients with bacterial pneumonia, mycoplasma pneumonia and streptococcal tonsillitis (n = 135). Results were compared with values obtained from 144 healthy controls. All biomarkers were elevated in bacterial and viral infections compared to healthy controls. Calprotectin was significantly increased in patients with bacterial infections; bacterial pneumonia, mycoplasma pneumonia and streptococcal tonsillitis compared with viral infections. PCT was significantly elevated in patients with bacterial pneumonia compared to viral infections but not in streptococcal tonsillitis or mycoplasma caused infections. HBP was not able to distinguish between bacterial and viral causes of infections. The overall clinical performance of calprotectin in the distinction between bacterial and viral respiratory infections, including mycoplasma was greater than performance of PCT and HBP. Rapid determination of calprotectin may improve the management of respiratory tract infections and allow more precise diagnosis and selective use of antibiotics.
Detection of the in vitro modulation of Plasmodium falciparum Arf1 by Sec7 and ArfGAP domains using a colorimetric plate-based assay
Authors: Swart Tarryn , Khan Farrah D. , Ntlantsana Apelele , Laming Dustin , Veale Clinton G. L. , Przyborski Jude M. , Edkins Adrienne L. , Hoppe Heinrich C.
Published: March 2020

The regulation of human Arf1 GTPase activity by ArfGEFs that stimulate GDP/GTP exchange and ArfGAPs that mediate GTP hydrolysis has attracted attention for the discovery of Arf1 inhibitors as potential anti-cancer agents. The malaria parasite Plasmodium falciparum encodes a Sec7 domain-containing protein - presumably an ArfGEF - and two putative ArfGAPs, as well as an Arf1 homologue (PfArf1) that is essential for blood-stage parasite viability. However, ArfGEF and ArfGAP-mediated activation/deactivation of PfArf1 has not been demonstrated. In this study, we established an in vitro colorimetric microtiter plate-based assay to detect the activation status of truncated human and P. falciparum Arf1 and used it to demonstrate the activation of both proteins by the Sec7 domain of ARNO, their deactivation by the GAP domain of human ArfGAP1 and the inhibition of the respective reactions by the compounds SecinH3 and QS11. In addition, we found that the GAP domains of both P. falciparum ArfGAPs have activities equivalent to that of human ArfGAP1, but are insensitive to QS11. Library screening identified a novel inhibitor which selectively inhibits one of the P. falciparum GAP domains (IC50 4.7 µM), suggesting that the assay format is suitable for screening compound collections for inhibitors of Arf1 regulatory proteins.
Molecular characteristics of the first case of haloxyfop-resistant Poa annua
Authors: Ghanizadeh H. , Mesarich C. H. , Harrington K. C.
Published: March 2020

Haloxyfop is one of two acetyl-coenzyme A carboxylase (ACCase) inhibitors that is recommended for controlling Poa annua. We have characterised a population of P. annua that had developed resistance to haloxyfop. This resistant population was found to be almost 20 times less sensitive to haloxyfop than a susceptible population based on percentage survival of individuals in two dose-response experiments. However, the haloxyfop-resistant population was still susceptible to clethodim. Pre-treatment of resistant individuals with a cytochrome P450 inhibitor, malathion, did not change the sensitivity level of the resistant plants to haloxyfop, suggesting that a non-target site mechanism of resistance involving enhanced metabolism, was not responsible for this resistance in P. annua. Gene sequencing showed that a target site mutation at position 2041, which replaced isoleucine with threonine in the carboxyltransferase (CT) domain of the ACCase enzyme, was associated with resistance to haloxyfop in the resistant population. An evaluation of the 3-D structure of the CT domain suggested that, unlike Asn-2041, which is the most common mutation at this position reported to date, Thr-2041 does not change the conformational structure of the CT domain. This is the first study investigating the molecular mechanism involved with haloxyfop resistance in P. annua.
Pathophysiological changes of the lower urinary tract behind voiding dysfunction in streptozotocin-induced long-term diabetic rats
Authors: Masuda Kazuki , Aizawa Naoki , Watanabe Daiji , Okegawa Takatsugu , Kume Haruki , Igawa Yasuhiko , Fukuhara Hiroshi
Published: March 2020

We evaluated pathophysiological characteristics of the lower urinary tract dysfunction in a streptozotocin (STZ)-induced diabetic rat model. STZ (60 mg/kg) was injected intraperitoneally into male Wistar rats. In vitro bladder muscle strip experiments, in vivo cystometry, and simultaneous recordings of bladder pressure + urethral perfusion pressure (BP + UPP) with or without intravenous administration of L-arginine (300 mg/kg) or tadalafil (0.03 mg/kg) were performed at several time points. In vitro muscle strip experiments demonstrated that diabetic rats had significantly higher contractile responses to carbachol at 4–16 weeks, and a tendency for higher contractile responses to electrical field stimulation at 4–12 weeks, but this was reversed at 16 weeks. Diabetic rats had significant increases in voided volume, residual volume, bladder capacity, maximal voiding pressure, and amplitude and frequency of non-voiding contractions at 16 weeks. Tadalafil decreased the residual volume in diabetic rats. Diabetic rats had significantly higher UPP nadir and mean UPP during high-frequency oscillation at 16 weeks, which were reversed by tadalafil or L-arginine administration. The present results suggest that urethral relaxation failure, probably related to impairment of the NO/cGMP signalling pathway, rather than bladder contractile dysfunction may be a prominent cause for voiding dysfunction in STZ-induced chronic diabetic rats.
Biochemical features of the novel Tail Tubular Protein A of Yersinia phage phiYeO3-12
Authors: Pyra Anna , Urbańska Natalia , Filik Karolina , Tyrlik Katherine , Brzozowska Ewa
Published: March 2020

Tail Tubular Protein A (TTPA) was long thought to be strictly a structural protein of environmental bacteriophages. However, our recent work has suggested that some TTPAs have additional functional features and thus are dual-function proteins. This study introduces a new TTPA family member, TTPAgp11, which belongs to Yersinia phage phiYeO3-12. We cloned the gene, expressed it and then purified the phage protein. The protein, including its hydrolytic activity, was characterized. Our enzymatic activity tests showed that TTPAgp11 displayed hydrolytic activity towards Red-starch, suggesting that this enzyme could be classified as part as the α − 1, 4-glucosidase family. Protein folding and aggregation tests indicated that TTPAgp11 is a single-domain protein whose aggregation can be induced by maltose or N-acetylglucosamine. The spatial structure of TTPAgp11 seemed to resemble that of the first reported dual-function TTPA, TTPAgp31, which was isolated from Klebsiella pneumoniae phage 32.
The position of the target site for engineered nucleases improves the aberrant mRNA clearance in in vivo genome editing
Authors: Lee Jae Hoon , Yu Sungsook , Nam Tae Wook , Roh Jae-il , Jin Young , Han Jeong Pil , Cha Ji-Young , Kim Yoon Ki , Yeom Su-Cheong , Nam Ki Taek , Lee Han-Woong
Published: March 2020

Engineered nucleases are widely used for creating frameshift or nonsense mutations in the target genes to eliminate gene functions. The resulting mRNAs carrying premature termination codons can be eliminated by nonsense-mediated mRNA decay. However, it is unclear how effective this process would be in vivo. Here, we found that the nonsense-mediated decay was unable to remove the mutant mRNAs in twelve out of sixteen homozygous mutant mice with frameshift mutations generated using engineered nucleases, which is far beyond what we expected. The frameshift mutant proteins translated by a single nucleotide deletion within the coding region were also detected in the p53 mutant mice. Furthermore, we showed that targeting the exons present downstream of the exons with a start codon or distant from ATG is relatively effective for eliminating mutant mRNAs in vivo, whereas the exons with a start codon are targeted to express the mutant mRNAs. Of the sixteen mutant mice generated, only four mutant mice targeting the downstream exons exhibited over 80% clearance of mutant mRNAs. Since the abnormal products, either mutant RNAs or mutant proteins, expressed by the target alleles might obscure the outcome of genome editing, these findings will provide insights in the improved performance of engineered nucleases when they are applied in vivo.
Cathepsin g Degrades Both Glycosylated and Unglycosylated Regions of Lubricin, a Synovial Mucin
Authors: Huang Shan , Thomsson Kristina A. , Jin Chunsheng , Alweddi Sally , Struglics André , Rolfson Ola , Björkman Lena I. , Kalamajski Sebastian , Schmidt Tannin A. , Jay Gregory D. , Krawetz Roman , Karlsson Niclas G. , Eisler Thomas
Published: March 2020

Lubricin (PRG4) is a mucin type protein that plays an important role in maintaining normal joint function by providing lubrication and chondroprotection. Improper lubricin modification and degradation has been observed in idiopathic osteoarthritis (OA), while the detailed mechanism still remains unknown. We hypothesized that the protease cathepsin G (CG) may participate in degrading lubricin in synovial fluid (SF). The presence of endogenous CG in SF was confirmed in 16 patients with knee OA. Recombinant human lubricin (rhPRG4) and native lubricin purified from the SF of patients were incubated with exogenous CG and lubricin degradation was monitored using western blot, staining by Coomassie or Periodic Acid-Schiff base in gels, and with proteomics. Full length lubricin (∼300 kDa), was efficiently digested with CG generating a 25-kDa protein fragment, originating from the densely glycosylated mucin domain (∼250 kDa). The 25-kDa fragment was present in the SF from OA patients, and the amount was increased after incubation with CG. A CG digest of rhPRG4 revealed 135 peptides and 72 glycopeptides, and confirmed that the protease could cleave in all domains of lubricin, including the mucin domain. Our results suggest that synovial CG may take part in the degradation of lubricin, which could affect the pathological decrease of the lubrication in degenerative joint disease.
Identification of the key genes and pathways involved in the tumorigenesis and prognosis of kidney renal clear cell carcinoma
Authors: Cui Hao , Shan Hongjian , Miao Michael Zhe , Jiang Zhiguo , Meng Yuanyuan , Chen Ran , Zhang Longzhen , Liu Yong
Published: March 2020

Kidney renal clear cell carcinoma (KIRC) is the most common renal cell carcinoma (RCC). However, patients with KIRC usually have poor prognosis due to limited biomarkers for early detection and prognosis prediction. In this study, we analysed key genes and pathways involved in KIRC from an array dataset including 26 tumour and 26 adjacent normal tissue samples. Weighted gene co-expression network analysis (WGCNA) was performed with the WGCNA package, and 20 modules were characterized as having the highest correlation with KIRC. The upregulated genes in the tumour samples are involved in the innate immune response, whereas the downregulated genes contribute to the cellular catabolism of glucose, amino acids and fatty acids. Furthermore, the key genes were evaluated through a protein-protein interaction (PPI) network combined with a co-expression network. The comparatively lower expression of AGXT, PTGER3 and SLC12A3 in tumours correlates with worse prognosis in KIRC patients, while higher expression of ALOX5 predicts reduced survival. Our integrated analysis illustrated the hub genes involved in KIRC tumorigenesis, shedding light on the development of prognostic markers. Further understanding of the function of the identified KIRC hub genes could provide deep insights into the molecular mechanisms of KIRC.
A Novel Fluorescent Reporter System Identifies Laminin-511/521 as Potent Regulators of Cardiomyocyte Maturation
Authors: Chanthra Nawin , Abe Tomoyuki , Miyamoto Matthew , Sekiguchi Kiyotoshi , Kwon Chulan , Hanazono Yutaka , Uosaki Hideki
Published: March 2020

Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) hold great promise for disease modeling and drug discovery. However, PSC-CMs exhibit immature phenotypes in culture, and the lack of maturity limits their broad applications. While physical and functional analyses are generally used to determine the status of cardiomyocyte maturation, they could be time-consuming and often present challenges in comparing maturation-enhancing strategies. Therefore, there is a demand for a method to assess cardiomyocyte maturation rapidly and reproducibly. In this study, we found that Myomesin-2 (Myom2), encoding M-protein, is upregulated postnatally, and based on this, we targeted TagRFP to the Myom2 locus in mouse embryonic stem cells. Myom2-RFP+ PSC-CMs exhibited more mature phenotypes than RFP- cells in morphology, function and transcriptionally, conductive to sarcomere shortening assays. Using this system, we screened extracellular matrices (ECMs) and identified laminin-511/521 as potent enhancers of cardiomyocyte maturation. Together, we developed and characterized a novel fluorescent reporter system for the assessment of cardiomyocyte maturation and identified potent maturation-enhancing ECMs through this simple and rapid assay. This system is expected to facilitate use of PSC-CMs in a variety of scientific and medical investigations.
Data-driven modelling and spatial complexity supports heterogeneity-based integrative management for eliminating Simulium neavei-transmitted river blindness
Authors: Michael Edwin , Smith Morgan E. , Singh Brajendra K. , Katabarwa Moses N. , Byamukama Edson , Habomugisha Peace , Lakwo Thomson , Tukahebwa Edridah , Richards Frank O.
Published: March 2020

Concern is emerging regarding the challenges posed by spatial complexity for modelling and managing the area-wide elimination of parasitic infections. While this has led to calls for applying heterogeneity-based approaches for addressing this complexity, questions related to spatial scale, the discovery of locally-relevant models, and its interaction with options for interrupting parasite transmission remain to be resolved. We used a data-driven modelling framework applied to infection data gathered from different monitoring sites to investigate these questions in the context of understanding the transmission dynamics and efforts to eliminateSimulium neavei- transmitted onchocerciasis, a macroparasitic disease that causes river blindness in Western Uganda and other regions of Africa. We demonstrate that our Bayesian-based data-model assimilation technique is able to discover onchocerciasis models that reflect local transmission conditions reliably. Key management variables such as infection breakpoints and required durations of drug interventions for achieving elimination varied spatially due to site-specific parameter constraining; however, this spatial effect was found to operate at the larger focus level, although intriguingly including vector control overcame this variability. These results show that data-driven modelling based on spatial datasets and model-data fusing methodologies will be critical to identifying both the scale-dependent models and heterogeneity-based options required for supporting the successful elimination ofS. neavei-borne onchocerciasis.
Characteristics and associated factors of physical activity-related injuries among university students in southern China
Authors: Cai Weicong , Tang Dongchun , Gao Yang , Yang Wenda , Chen Shangmin , Gao Lijie , Jia Cunxian , Li Liping
Published: March 2020

This cross-sectional study aimed to describe the characteristics of physical activity-related injury (PARI) and to explore its associated factors among university students in Chaoshan district. Selected from the baseline survey in March and April, 434 students graded 1–3 from two universities were interviewed face-to-face in April and May 2017. Socio-demographics, physical activity (PA) participation, risk-taking behaviors, and PARI occurrences in the past 12 months were collected. Group Lasso logistic regression was applied to identify the risk factors of PARI. Totally, 317 PARI episodes were reported by 184 subjects with an overall injury risk of 0.73 injuries/student/year (males: 1.00, females: 0.63) and an injury incidence density of 0.81 injuries per 1000 PA exposure hours (males: 1.13, females: 0.69). Most injuries involved the lower extremities and were sprains and strains. Males, sports team members, and those with high-risk rebellious and anti-social behaviors were more likely to sustain PARI compared to their counterparts. Those who participated in vigorous-intensity PA with longer duration (particularly ≥ 150 min/week) were at a higher risk for PARI. This study indicates that PARI is a health concern among university students and great efforts should be taken to prevent them from PARI when promoting a physically active lifestyle.
Evolutionary and Molecular Characterization of liver-enriched gene 1
Authors: Dang Yanna , Wang Jin-Yang , Liu Chen , Zhang Kun , Jinrong Peng , He Jin
Published: March 2020

Liver-enriched gene 1 (Leg1) is a newly identified gene with little available functional information. To evolutionarily and molecularly characterize Leg1 genes, a phylogenetic study was first conducted, which indicated that Leg1 is a conserved gene that exists from bacteria to mammals. During the evolution of mammals, Leg1s underwent tandem duplications, which gave rise to Leg1a, Leg1b, and Leg1c clades. Analysis of the pig genome showed the presence of all three paralogs of pig Leg1 genes (pLeg1s), whereas only Leg1a could be found in the human (hLeg1a) or mouse (mLeg1a) genomes. Purifying force acts on the evolution of Leg1 genes, likely subjecting them to functional constraint. Molecularly, pLeg1a and its coded protein, pig LEG1a (pLEG1a), displayed high similarities to its human and mouse homologs in terms of gene organization, expression patterns, and structures. Hence, pLeg1a, hLeg1a, and mLeg1a might preserve similar functions. Additionally, expression analysis of the three Leg1as suggested that eutherian Leg1as might have different functions from those of zebrafish and platypus due to subfunctionalization. Therefore, pLeg1a might provide essential information about eutherian Leg1a. Moreover, a preliminary functional study using RNA-seq suggested that pLeg1a is involved in the lipid homeostasis. In conclusion, our study provides some basic information on the aspects of evolution and molecular function, which could be applied for further validation of Leg1 using pig models.
Fluctuations of the elemental composition in the layers of mineral deposits formed on the elements of biogas engines
Authors: Stanuch Ireneusz , Sozańska Maria , Biegańska Jolanta , Cebula Jan , Nowak Jacek
Published: March 2020

The aim of the research was to compare the chemical composition and morphology of the surface of the backsheet growing on the metallic and surface substrate growing already on the prepared mineral substrate. Moreover the aim of this study was also to analyze the dispersion of chemical elements on the cross-sections of deposits formed on parts of landfill biogas-driven engines. The chemical composition of extreme layers of mineral deposits extracted from the engine piston and their cross-sections from four pistons and one head was examined by SEM-EDS. The bottom side showed a much smaller heterogeneity of the topography of the surface than the topside. The chemical composition of the deposit bottom side it primarily S (41%), Si (34%) and Al (17%). In the case of the top side, the dominated of Ca (52%) with a relatively high share of S (32%) and Si (14%). The presence of P and Mg it was also found, but only on the bottom side, and the share of Fe and Zn only on the top side. In the case of cross-sections, Ca, S and Si were the dominant elements. In general, there were higher Si participations in the zone of the bottom layer with a downward trend to the top sheet. The mass shares of S and Ca were lower in the zone of the bottom layer with the upward trend to the top sheet, also undergoing fluctuation.
3-D magnetohydrodynamic AA7072-AA7075/methanol hybrid nanofluid flow above an uneven thickness surface with slip effect
Authors: Tlili Iskander , Nabwey Hossam A. , Ashwinkumar G. P. , Sandeep N.
Published: March 2020

A 3-D magnetohydrodynamic flow of hybrid nanofluid across a stretched plane of non-uniform thickness with slip effects is studied. We pondered aluminum alloys of AA7072 and AA7072 + AA7075 in methanol liquid. The aluminum alloys amalgamated in this study are uniquely manufactured materials, possessing enhanced heat transfer features. AA7072 alloy is a composite mixture of Aluminum & Zinc in the ratio 98 & 1 respectively with added metals Silicon, ferrous and Copper. Equally, AA7075 is a mixture of Aluminum, Zinc, Magnesium, and Copper in the ratio of ~90, ~6, ~3 and ~1 respectively with added metals Silicon ferrous and Magnesium. Numerical solutions are attained using R-K based shooting scheme. Role of physical factors on the flow phenomenon are analyzed and reflected by plots and numerical interpretations. Results ascertain that heat transfer rate of the hybrid nanoliquid is considerably large as matched by the nanofluid. The impact of Lorentz force is less on hybrid nanofluid when equated with nanofluid. Also, the wall thickness parameter tends to improve the Nusselt number of both the solutions.
Patient- and 3D morphometry-based nose outcomes after skeletofacial reconstruction
Authors: Denadai Rafael , Chou Pang-Yun , Seo Hyung Joon , Lonic Daniel , Lin Hsiu-Hsia , Pai Betty C. J. , Lo Lun-Jou
Published: March 2020

Patient satisfaction with the shape and appearance of their nose after orthognathic surgery-based skeletofacial reconstruction is an important, but often overlooked, outcome. We assessed the nose-related outcomes through a recently developed patient-reported outcome instrument and a widely adopted 3D computer-based objective outcome instrument, to verify any correlation in the results produced by these tools. We collected FACE-Q nose appearance reports (2 scales) and 3D nasal morphometry (10 parameters) from patients with class III skeletal pattern and congenital cleft lip palate deformity (n = 23) or developmental dentofacial deformity (n = 23) after (>12 months) skeletofacial reconstruction. The cleft and dentofacial cohorts demonstrated significantly (p < 0.001) poorer satisfaction scores with regard to the FACE-Q nostrils scale than the normal age-, gender-, and ethnicity-matched subjects (n = 107), without any significant difference in FACE-Q nose scale. The cleft cohort had significantly (p < 0.001) smaller nasal length, nasal tip projection, and columellar angle and greater nasal protrusion, alar width, and columellar–labial angle values than the dentofacial and normal cohorts; however, there were no significant differences between the dentofacial versus normal cohorts. The FACE-Q nose and nostrils scales were significantly (p < 0.001; r = −0.26–0.27) correlated to the results of the 3D morphometric analysis, with regard to nasal length, alar width, columella angle, and columellar–labial angle parameters. This study revealed differences in satisfaction with the appearance of the nose according to the type of underlying deformity, and demonstrated a significant correlation (low correlation coefficients) between the patient-reports and 3D image-based outcome measure tools, which has implications for multidisciplinary-centered research, auditing, and clinical care.
Effect of root interaction on nodulation and nitrogen fixation ability of alfalfa in the simulated alfalfa/triticale intercropping in pots
Authors: Zhao Yajiao , Liu Xiaojing , Tong Changchun , Wu Yong
Published: March 2020

Cereal/legume intercropping is likely to achieve the optimal exploitation of soil and atmospheric nitrogen (N) sources to maintain high production and quality levels with low N inputs, as an attempt to eliminate underlying environmental effects. Nevertheless, the extent of the effect of cereal/legume intercropping on nodulation and N fixation of intercropped legumes in root interaction requires extensive verification. In the present study, root interaction of alfalfa/triticale intercropping was simulated in pots with the use of root separation types (pot with no barrier (A-T), pot with nylon mesh barrier (NA-T), pot with plastic barrier (PA-T), and alfalfa alone (SA)) in pots. Moreover, the experiment was measured at a range of N levels (N21, N210) and growing stages (branching, budding and initial flowering stages) in growth chamber. As alfalfa was growing, the total nodule number (TNN), effective nodule number (ENN) and nitrogenase activity (NA) of alfalfa with A-T and other cropping systems more noticeably differed from each other at higher N levels, whereas their diversification was reduced at lower N levels. As alfalfa was growing continuously, fresh nodule weight per plant (PNW) and single fresh nodule weight (SNW) with A-T and other cropping systems were amplified more significantly. The nodulation and N fixation ability under N21 were more significant than those under N210. Dry weight of plant per pot (TDW) and nitrogen accumulation of plant per pot (TNA) with A-T were obviously higher than those with other systems in the initial flowering stage, except for TNA under N21. The parameters regarding the nodulation and N fixation ability were significantly positively correlated on the whole. However, SNW and TNA were not significantly correlated, neither were SNW and TDW. According to the mentioned results, the closer root interaction, the better the nodulation form and N fixation ability of alfalfa will be, and the higher the biomass and N accumulation of all plants in pots will be. Interspecific facilitation in alfalfa/triticale intercropping system resulted in a greater yield and N accumulation; it also ultimately enhanced nodulation and N fixation ability, which can be applied in sustainable systems to avoid N loss to the environment and enhance N use efficiency.
Bell’s palsy and obesity, alcohol consumption and smoking: A nested case-control study using a national health screening cohort
Authors: Kim So Young , Oh Dong Jun , Park Bumjung , Choi Hyo Geun
Published: March 2020

The aim of this study was to investigate the association of body mass index (BMI), alcohol consumption, and smoking status with the occurrence of Bell’s palsy. The Korean National Health Insurance Service-Health Screening Cohort of a ≥ 40-year-old population from 2000–2003 was used. A total of 5,632 Bell’s palsy participants were matched with 22,528 control participants in terms of age, sex, income, region of residence, and past medical histories of hypertension, diabetes, and dyslipidemia. Bell’s palsy was classified by a history of ≥2 diagnoses with ICD-10 code (G510) and steroid treatment. BMI (kg/m2) was classified as <18.5 (underweight), ≥18.5 to <23 (normal), ≥23 to <25 (overweight), ≥25 to <30 (obese I), and ≥30 (obese II). Alcohol consumption was divided into non-drinkers and those who drank 2–3 times a month, 1–2 times a week, and ≥3 times a week. Smoking status was categorized as current smokers, past smokers, and non-smokers. The odds of obesity, alcohol consumption, and smoking with Bell’s palsy were analyzed using logistic regression analysis. BMI showed proportionally positive associations with Bell’s palsy (adjusted OR [95% CI] = 0.61 [0.47–0.79] for underweight, 1.16 [1.08–1.26] for normal, 1.24 [1.15–1.33] for obese I, and 1.61 [1.38–1.88] for obese II, P < 0.001). The odds of alcohol consumption with Bell’s palsy were 0.90 (95% confidence interval [CI] = 0.82–0.99) for 2–3 times a month, 0.77 (95% CI = 0.69–0.85) for 1–2 times a week, and 0.79 (95% CI = 0.71–0.88) for ≥3 times a week compared to nondrinkers (P < 0.001). Smoking did not show a relationship with the occurrence of Bell’s palsy. Obesity was related to the risk of Bell’s palsy in the population over 40 years old. On the other hand, alcohol consumption was negatively associated with the occurrence of Bell’s palsy.