bluuebunny/crossref_metadata_2025_split huggingface dataset:
WITHDRAWN: Glioblastoma cell-derived extracellular vesicle miR-27a-3p facilitates M2 macrophage polarization
Authors: Zhao Guifang , Yu Hongquan , Ding Lijuan , Wang Weiyao , Wang Huang , Hu Yao , Qin Lingsha , Deng Guangce , Xie Buqing , Li Guofeng , Qi Lin 
Published: May 2020

Abstract
Background: M2 macrophage polarization has been found to be correlated with malignancy of glioblastoma. In this study, we investigated the potential role of microRNA (miRNA) derived from extracellular vesicles of glioblastoma (glioblastoma-EVs) in M2 macrophage polarization.Methods: After isolation of human glioblastoma-EVs, transmission electron microscopy (TEM) and Nano-particle tracking analysis (NTA) were performed to identify the EVs. Besides, the proliferation, migration and invasion of glioma cells were analyzed by CCK8 and Transwell assays, respectively. The target genes of miR-27a-3p were predicted through bioinformatics analysis and verified by dual-luciferase reporter gene assay. ChIP assay was applied to detect the binding of enhancer of zeste homologue 1 (EZH1) to lysine-specific demethylase 3A (KDM3A) promoter region and the interaction between KDM3A and connective tissue growth factor (CTGF). Glioblastoma mouse models were established followed by the implement of hematoxylin-eosin (HE) and ELISA staining on pathological changes of mouse brain tissues.Results: Human glioblastoma-EVs were successfully isolated. The high expression of miR-27a-3p was found in glioblastoma tissues as well as glioblastoma-EVs, which could induce M2 polarization, thus promoting glioblastoma cell proliferation, migration and invasion. It was also shown that miR-27a-3p targeted EZH1 and promoted KDM3A expression to elevate the expression of CTGF. Glioblastoma-EVs delivered miR-27a-3p to promote the KDM3A-upregulated CTGF by downregulating EZH1, thereby promoting M2 macrophage polarization and development of glioblastoma in vivo.Conclusion: These findings highlight that EV miR-27a-3p may promote M2 macrophage polarization, which is associated with the progression of tumors.
Intein-mediated intracellular production of active microbial transglutaminase in Corynebacterium glutamicum
Authors: Zhang Nan , Zhang Shan , He Yongzhi , Chen Xin , Zhang Yanfeng , Dong Zhiyang 
Published: May 2020

Abstract
Background The microbial transglutaminase (mTGase) from Streptomyces mobaraense has been widely used in the food industry. Recombinant production of mTGase is tricky because the mTGase is synthesized as an inactive zymogen, which needs to be activated by proteolytic processing. Self-cleaving inteins have been applied to activate the zymogen in a simple and highly specific manner as compared with proteolytic processing. However, self-cleaving inteins suffer from the inherent problem of premature cleavage. Moreover, self-cleaving inteins normally require an additional step of long time incubation to induce the cleavage. These two inherent problems limit self-cleaving inteins for their potential application in the production of mTGase.Results In this study, the premature cleavage of intein Ssp DnaB was observed in Corynebacterium glutamicum when the Ssp DnaB was used to activate mTGase precursor protein. Rather than suppressing it, the premature cleavage was applied to produce active mTGase in C. glutamicum. The SDS-PAGE analysis and the mTGase activity assay indicated that the premature cleavage of intein Ssp DnaB was successfully applied to activate the mTGase intracellularly in C. glutamicum. The subsequent N-terminal amino acid sequencing and site-directed mutagenesis studies demonstrated that the premature cleavage activated the mTGase intracellularly in a highly specific manner. Finally, in a jar fermentor, the intracellular mTGase activity was up to 49 U/mL, which was the highest intracellular mTGase activity ever reported.Conclusions An efficient and simple approach with great potential for large-scale industrial production of active mTGase was presented in this study. This approach employed premature cleavage of intein Ssp DnaB to activate mTGase in C. glutamicum, resulting in high-level intracellular production of active mTGase. Moreover, this approach did not require any further processing steps such as protease treatment or long time incubation, greatly simplifying the production of active mTGase.
Production of bio-xylitol from D-xylose by an engineered Pichia pastoris expressing a recombinant xylose reductase did not require any auxiliary substrate as electron donor
Authors: Louie Michael , Louie Kailin , DenHartog Samuel , Gopishetty Sridhar , Arnold Mark , Das Shuvendu 
Published: May 2020

Abstract
Background: Xylitol is a five-carbon sugar alcohol that has numerous beneficial health properties. It has almost the same sweetness as sucrose but has lower energy value compared to the sucrose. Metabolism of xylitol is insulin independent and thus it is an ideal sweetener for diabetics. It is widely used in food products, oral and personal care, and animal nutrition as well. Here we present a two-stage strategy to produce bio-xylitol from D-xylose using a recombinant Pichia pastoris expressing a heterologous xylose reductase gene. The recombinant P. pastoris cells were first generated by a low-cost, standard procedure. The cells were then used as a catalyst to make the bio-xylitol from D-xylose.Results: P. pastoris expressing XYL1 from P. stipitis and gdh from B. subtilis demonstrated that the biotransformation was very efficient with as high as 80% (w/w) conversion within two hours. The whole cells could be re-used for multiple rounds of catalysis without loss of activity. Also, the cells could directly transform D-xylose in a non-detoxified hemicelluloses hydrolysate to xylitol at 70% (w/w) yield.Conclusions: We demonstrated here that the recombinant P. pastoris expressing xylose reductase could transform D-xylose, either in pure form or in crude hemicelluloses hydrolysate, to bio-xylitol very efficiently. This biocatalytic reaction happened without the external addition of any NAD(P)H, NAD(P)+, and auxiliary substrate as an electron donor. Our experimental design & findings reported here are not limited to the conversion of D-xylose to xylitol only but can be used with other many oxidoreductase reactions also, such as ketone reductases/alcohol dehydrogenases and amino acid dehydrogenases, which are widely used for the synthesis of high-value chemicals and pharmaceutical intermediates.
Microbial characteristics of the DMFT index in a high HIV prevalence setting
Authors: Kalanzi Dunstan , Kizza Harriet Mayanja , Nakanjako Damalie , Mboowa Gerald , Mbabali Muhammad , Kigozi Edgar , Ashaba Fred , Sserwadda Ivan , Kateete David Patrick , Acan Beatrice , Sewankambo Nelson K , Muwonge Adrian 
Published: May 2020

Abstract
Background: Oral disease pathogenesis is primarily driven by microbial dysbiosis although diagnosis is routinely macroscopic. To improve early detection especially in HIV patients who are disproportionately affected, there is need to reconcile macroscopic and microscopic characteristics of disease. This study aimed to use amplicon sequencing data to characterize oral microbiota changes along the decayed, missing, filled teeth (DMFT) index.Methods: Amplicon sequencing of the V6-V8 region of the 16S rRNA gene was done on DNA recovered from whole unstimulated saliva of 59 HIV positive and 29 HIV negative individuals, respectively. The microbial structure, composition and co-occurrence networks were characterized using QIIME-2, Phyloseq, Microbiome-1.9.2 and Metacoder in R.Results: From a total of 2.6 million high quality sequence reads obtained, we characterized the oral microbiota into 2,093 operational taxonomic units (OTUs), 21 phyla and 239 genera. While oral microbiota did not cluster participants into distinct groups that track with the DMFT index, we observed the following: a) A steady increase in accessory microbiota while the core size (~50% of richness) remained stable. b) The abundance of core genera such as Stomatobaculum, Peptostreptococcus and Campylobacter increased at onset of pathology (low DMFT), c) A general increase in oral microbial biomass with a typical log difference between gingivitis and periodontitis. d) The onset of pathology (low DMFT) was associated with massive reduction in oral microbial entropy.Conclusions: Although oral microbial shifts along the DMFT index are not distinct, we have demonstrated the potential utility of microbiota dynamics to inform oral disease characteristics. We therefore propose a framework to inform future clinical oral metagenomic studies especially among HIV positive persons in resource limited settings.
Dental Students’ Notetaking: a Thematic, Material-based Interview Study
Authors: Lindberg Viveca , Jounger Sofia Louca , Christidis Maria , Christidis Nikolaos 
Published: May 2020

Abstract
Background
The transition from upper secondary to higher education and from higher education to professional practice require that students need to adapt to new literacy practices, academic and professional. The aim of this study was to map what characterizes dental students’ notetaking and secondarily what dental students express regarding their notetaking.
Methods
Three undergraduate dental students participated in this material-based, semi-structured interview study, that was framed within a New Literacy Studies approach. To analyze the outcome of the interviews a thematic analysis was used.
Results
Eight themes that relate to what, how and for what purpose students write were discerned. These include a) what: T1-Professional vocabulary; T2-Core content; T3-Clinical example; b) how: T4-Multimodal accentuation; T5-Synthesis; and c) for what purpose: T6-Mnemonic strategies; T7-Academic purposes; and T8 professional purposes.
Conclusions
The digital development has changed the notetaking which has evolved from just synchronous to become both multimodal and asynchronous and is more of a writing practice. Students’ writing practices seem to be motivated by their knowledge formation in relation to a subject matter. Results showed that students, half-way through their dental education, are aware of that notetaking is not only for passing exams but also for learning for their profession.
Hypoxia-inducible factor-2α but not hypoxia-inducible factor-1α increases iron traffic in astrocytes
Authors: Xu Manman , Chen Zhiqiang , Ma Xizhen , Xu Hangrui , Fan Changsheng , Xie Junxia , Wang Jun 
Published: May 2020

Abstract
Background: Disrupted iron homeostasis in the substantia nigra (SN) is an important mechanism in Parkinson’s disease (PD). In our previous studies using cultured neurons, we showed that 6-hydroxydopamine (6-OHDA) treatment increased the expression of divalent metal transporter-1 (DMT1) gating iron influx and decreased the expression of ferroportin 1 (FPN1) gating iron efflux, leading to increased iron deposition. In astrocytes, 6-OHDA increased both DMT1 and FPN1 expressions and iron traffic. However, the underlying mechanisms mediating these processes remain elusive. Hypoxia-inducible factors (HIFs) are important regulators of iron homeostasis.Methods: Using western blots, we observed HIFs, DMT1 and FPN1 expressions in primary cultured astrocytes and ventral mesencephalic (VM) treated with 6-OHDA, inhibitors of HIF-1α and HIF-2α, protein kinase C (PKC) inhibitor and PKC activator, radical scavenger and inducible NO synthase (iNOS) inhibitor . The ferrous iron traffic of astrocytes was determined by measuring the quenching or reversing of calcein fluorescence.Results: Using primary rat cell cultures, we observed that 6-OHDA treatment significantly increased the expressions of HIF-1α and HIF-2α in cultured astrocytes but not in VM neurons. Moreover, we observed that HIF-2α inhibitor, but not HIF-1α inhibitor reversed the 6-OHDA-induced upregulation of DMT1 and FPN1 and ferrous iron traffic. Blocking protein kinase C (PKC) pathway by bisindolylmaleimide I hydrochloride (Bisl) or N-acetyl-l-cysteine (NAC) or Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) inhibits Phorbol 12-myristate 13-acetate (PMA) or 6-OHDA induced activation of HIF-2α, whereas activating by PMA or 6-OHDA was sufficient to activate HIF-2α. Notably, blocking of PKC delta (PKC δ) phosphorylation by NAC, or L-NAME inhibits 6-OHDA induced activation of HIF-2α.Conclusion: Our data indicate that astrocytic HIF-2α, but not HIF-1α, may be an important regulator of DMT1 and FPN1 expressions by activating the PKC -dependent pathway.
Diesel Exhaust Particle Exposure Reduces Expression of the Epithelial Tight Junction Protein Tricellulin
Authors: Smyth Timothy , Veazey Janelle , Eliseeva Sophia , Chalupa David , Elder Alison , Georas Steve 
Published: May 2020

Abstract
Background
While exposure to diesel exhaust particles has been linked to aberrant immune responses in allergic diseases such as asthma, little attention has been payed to their effects on the airway epithelium. In this study, we sought to determine the effect of diesel exhaust exposure on airway epithelial barrier function and composition using in vitro and in vivo model systems. 16HBE14o- human bronchial epithelial cells were grown on collagen coated Transwell inserts and exposed to 5 to 50 µg/cm2 SRM 2975 diesel particulate matter (DEP) suspended in cell culture medium or vehicle controls. Changes in barrier function were assessed by measuring transepithelial electrical resistance (TEER) and permeability to 4 kDa FITC Dextran. Neonatal BALB/c mice were exposed to aerosolized DEP (255 ± 89 µg/m3; 2 hours per day for 5 days) and changes in the tight junction protein Tricellulin were assessed two weeks post exposure.
Results
A six-hour incubation of epithelial cells with diesel exhaust particles caused a significant concentration-dependent reduction in epithelial barrier integrity as measured by decreased TEER and increased permeability to 4 kDa FITC-Dextran. This reduction in epithelial barrier integrity corresponded to a significant reduction in expression of the tight junction protein Tricellulin. siRNA mediated knockdown of Tricellulin recapitulated changes in barrier function caused by DEP exposure. Neonatal exposure to aerosolized DEP caused a significant reduction in lung Tricellulin two weeks post exposure at both the protein and mRNA level.
Conclusion
Short term exposure to DEP causes a significant reduction in epithelial barrier integrity through a reduction in the tight junction protein Tricellulin. Neonatal exposure to aerosolized DEP caused a significant and sustained reduction in Tricellulin protein and mRNA in the lung, suggesting that early life exposure to inhaled DEP may cause lasting changes in airway epithelial barrier function.
Fear of missing out (FoMO) and gaming disorder among Chinese university students: Impulsivity and game time as mediators
Authors: Li Li , Griffiths Mark D. , Niu Zhimin , Mei Songli 
Published: May 2020

Abstract
Background and aims: Research into the fear of missing out (FoMO) has greatly increased in recent years. Given the negative consequences of gaming disorder (GD) among a small minority of individuals, there is an increasing need for research examining the impact of FoMO on GD. However, little is known about the roles of impulsivity and gaming time as mediators in the relationship between FoMO and GD. The present study examined whether impulsivity and gaming time mediated the relationship between FoMO (trait-FoMO and state-FoMO) and GD among Chinese university students, as well as the prevalence of GD.Methods: A total of 1127 university students completed an online survey including the Chinese Trait-State Fear of Missing Out Scale (T-SFoMOS-C), the Barratt Impulsiveness Scale-Brief (BIS-Brief), gamine time survey, and the Chinese Gaming Disorder Scale (CGDS).Results: The prevalence of GD was 6.4% among Chinese university students. Trait-FoMO was found to indirectly impact GD via impulsivity and gamine time, whereas the direct effect of trait-FoMO on GD and the mediation effects of gaming time were not confirmed. State-FoMO impacted on GD both directly, and indirectly via the mediation effects of impulsivity as well as impulsivity and gaming time.Conclusion: Trait-FoMO on GD was fully mediated via impulsivity and gaming time, whereas state-FoMO on GD was partly mediated via impulsivity and gaming time. Individuals with high levels of FoMO were more likely to show impulsivity and spent a longer time gaming, and was associated with GD. These findings provide insights to incorporate into health prevention programs to help regulate emotion, control impulsivity, and decrease GD.
The mutational load and a T-cell inflamed tumor phenotype identify ovarian cancer patients rendering tumor-reactive T cells from PD-1+ tumor-infiltrating lymphocytes
Authors: Salas-Benito Diego , Conde Enrique , Tamayo-Uria Ibon , Mancheño Uxua , Elizalde Edurne , Garcia-Ros David , Aramendia Jose M. , Muruzabal Juan C. , Alcaide Julia , Guillen-Grima Francisco , Minguez Jose A. , Amores-Tirado Jose , Gonzalez-Martin Antonio , Sarobe Pablo , Lasarte Juan J. , Ponz-Sarvise Mariano , Andrea Carlos E. De , Hervas-Stubbs Sandra 
Published: May 2020

Abstract
Background: Adoptive immunotherapy with tumor-infiltrating lymphocytes (TIL) may benefit from the use of selective markers, such as programmed cell death protein 1 (PD-1), for tumor-specific T-cell enrichment, as well as predictive biomarkers that help identify those patients capable of rendering tumor-reactive TIL products. We have investigated this in ovarian cancer (OC) patients as candidate for TIL therapy implementation. Methods: PD-1- and PD-1+ CD8 TILs were isolated from resected ovarian tumors and, after polyclonal expansion, TIL products were tested against autologous tumor cells. Reactivity was assessed by IFNg production (ELISPOT) and upregulation of CD137. Baseline tumor samples were examined using flow cytometry, multiplexed quantitative immunofluorescence, Nanostring technology, for gene expression profile (GEP) analyses, as well as a next generation sequencing gene panel, for tumor mutational burden (TMB) calculation, to identify those features that distinguished patients with tumor-reactive and non-tumor-reactive TIL products.Results: Tumor-reactive TILs were detected in half of patients and were exclusively present in cells derived from the PD-1+ fraction. Flow-cytometric studies revealed that fresh tumors from patients rendering tumor-reactive TILs presented a significantly higher frequency of CD137+ cells within the PD1+CD8+ subset. Multiplexed immunofluorescence supported this finding, which was particularly striking in intraepithelial CD8 TILs. Baseline GEP analysis showed that patients rendering tumor-reactive TILs exhibited a significantly higher T-cell inflamed signature. Despite no correlation between TMB and GEP, both parameters stratified tumors, with patients with higher TMB and/or T-cell inflamed signature score rendering tumor-reactive TILs. Conclusion: We have demonstrated that PD-1 identifies autologous-tumor specific CD8 T cells infiltrating ovarian tumors and have uncovered predictive factors that identify OC patients who are likely to render tumor-reactive cells from PD-1+ TILs. These findings have important implications for improving the efficacy of TIL therapy in OC.
“We try our best to offer them the little that we can” Coping Strategies of Ghanaian Community Psychiatric Nurses: A Qualitative Study
Authors: Opare Frederick Yaw , Aniteye Patience , Afaya Agani , Glover-Meni Nathaniel 
Published: May 2020

Abstract
Introduction Addressing strategies of coping among community psychiatric nurses (CPNs) could help increase an individual’s ability to cope effectively and as a result, reduce experienced levels of stress. It is known that the level of work stress experienced by CPNs is remarkably high, because psychiatric nurses face more intense relationships with their patients, trying to prevent self-harm, and most at times face challenging situations with aggressive patient behaviors in the working environment. Therefore, the study aimed to explore the coping strategies employed by CPNs during healthcare delivery in the community. Methods An exploratory qualitative study design using an interpretative approach was employed. A total of 13 CPNs were purposively sampled, and data gathered through individual interviews using an interview guide. The interviews were audio-taped, transcribed verbatim and analyzed thematically, describing some of the coping strategies employed by CPNs in Ghana. Results Participants’ ages ranged from 26-60years. They comprised 3 males and 10 females. An inductive thematic analysis identified the following major coping strategies of community psychiatric nurses; self-disguise (wearing of mufti), reliance on religious faith, self-motivation and reduction in the number of home visits. Conclusion Coping strategies among community psychiatric nurses are essential for both individuals and organisations. Thus, community psychiatric nurses must acquire personal-mastery coping strategies such as the reduction in the number of home visits, self-disguise (wearing mufti) the use of faith to motivate themselves in the face the challenging working environment. The coping strategies adopted by community psychiatric nurses was not to only help them deliver care, but also as a way of protecting the clients so that people would not tag them as ‘mental patients’
Phone-based audience response system as an adjunct in orthodontic teaching of undergraduate dental students: a cross-over randomised controlled trial
Authors: Alharbi Fahad , Alazmi Khulud , Momani Bashar R. El , Almukhtar Anas , Al-Muzian Lubna , Almuzian Mohammed 
Published: May 2020

Abstract
Background: The advent of electronic teaching facilities opened doors towards a better tutor-student communication. The aim of this study is to explore the effectiveness of phone-based ARS (PB-ARS) using text- and multimedia-based questions on information retention by male dental undergraduate students and to explore the students’ perception and acceptance of ARS. Methods: A cross-over clustered randomised control trial was conducted at the College of Dentistry, Prince Sattam Bin Abdulaziz University/Alkharj/ Saudi Arabia. Students were allocated to one of two event groups (G1 or G2) using computer generated randomization. During the first intervention (lecture 1), G1 received ARS assisted lecture and G2 acts as a control group, then crossed over during the second intervention (lecture 2). Data for short- and long-term information retention were treated on intention to treat analysis basis and tested using Mann–Whitney U and t-test in addition to qualitative assessment of students’ perception and acceptance. Results : The results showed marginal improvements in short and long term retention of information in addition to positive attitude of students towards the ARS system. 87.5% of students in ARS group showed an improvement in their quiz score compared to 79.3% for control group. No statistically significant differences were observed between the intervention and control groups. Conclusion : the ARS system could be considered as an adjunctive teaching method to improve on information retention and interest in Orthodontics taught materials.