bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic characterization of a Marek’s disease virus strain isolated in Japan
Authors: Murata Shiro , Machida Yuka , Isezaki Masayoshi , Maekawa Naoya , Okagawa Tomohiro , Konnai Satoru , Ohashi Kazuhiko
Published: July 2020

Background: Marek’s disease virus (MDV) causes malignant lymphomas in chickens (Marek’s disease, MD). MD is currently controlled by vaccination; however, MDV strains have a tendency to develop increased virulence. Distinct diversity and point mutations are present in the Meq proteins, the oncoproteins of MDV, suggesting that changes in protein function induced by amino acid substitutions might affect MDV virulence. We previously reported that recent MDV isolates in Japan display distinct mutations in Meq proteins from those observed in traditional MDV isolates in Japan, but similar to those in MDV strains isolated from other countries. Methods: To further investigate the genetic characteristics in Japanese field strains, we sequenced the whole genome of an MDV strain that was successfully isolated from a chicken with MD in Japan. A phylogenetic analysis of the meq gene was also performed.Results: Phylogenetic analysis revealed that the Meq proteins in most of the Japanese isolates were similar to those of Chinese and European strains, and the genomic sequence of the Japanese strain was classified into the Eurasian cluster. Comparison of coding region sequences among the Japanese strain and MDV strains from other countries revealed that the genetic characteristics of the Japanese strain were similar to those of Chinese and European strains. Conclusions: The MDV strains distributed in Asian and European countries including Japan seem to be genetically closer to each other than to MDV strains from North America. These findings indicate that the genetic diversities of MDV strains that emerged may have been dependent on the different vaccination-based control approaches.
Auxin, abscisic acid and jasmonate are the central players in rice sheath rot caused by Sarocladium oryzae and Pseudomonas fuscovaginae
Authors: Peeters Kaat , Ameye Maarten , Demeestere Kristof , Audenaert Kris , Hofte Monica
Published: July 2020

Sheath rot is an emerging rice disease that causes severe yield losses worldwide. The main causal agents are the toxin producers Sarocladium oryzae and Pseudomonas fuscovaginae. The fungus S. oryzae produces helvolic acid and cerulenin and the bacterium P. fuscovaginae produces cyclic lipopeptides. Helvolic acid and the lipopeptide, fuscopeptin, inhibit membrane-bound H+-ATPase pumps in the rice plant. To manage rice sheath rot, a better understanding of the host response and virulence strategies of the pathogens is required. This study investigated the interaction of the sheath rot pathogens with their host and the role of their toxins herein. Japonica rice was inoculated with high- and low-helvolic acid-producing S. oryzae isolates or with P. fuscovaginae wild type and fuscopeptin mutant strains. During infection, cerulenin, helvolic acid and the phytohormones abscisic acid, jasmonate, auxin and salicylic acid were quantified in the sheath. In addition, disease severity and grain yield parameters were assessed. Rice plants responded to high-toxin-producing S. oryzae and P. fuscovaginae strains with an increase in abscisic acid, jasmonate and auxin levels. We conclude that, for both pathogens, toxins play a core role during sheath rot infection. S. oryzae and P. fuscovaginae interact with their host in a similar way. This may explain why both sheath rot pathogens cause very similar symptoms despite their different nature.
Gender Modifies the Effect of Body Mass Index on Lung Function Decline in Mild-to-Moderate COPD Patients: A Pooled Analysis
Authors: Chen Wenjia , Sadatsafavi Mohsen , FitzGerald J Mark , Lynd Larry , Sin Don
Published: July 2020

Background: Body weight is a poor prognostic risk factor in patients with chronic obstructive pulmonary disease (COPD). However, it is not known whether gender modifies this relationship.Methods: We pooled data of 8,686 COPD patients from 7 studies. Using a longitudinal natural cubic spline regression model, we examined the dose-response relationship between body mass index (BMI) and the rate of decline in forced expiratory volume in one second (FEV1) in patients with GOLD 1 and 2 disease, stratified by gender and adjusted for smoking status and cohort effects. Results: There was an inverse, nearly relationship between BMI and the rate of FEV1 decline in GOLD Grades 1 and 2, which was modified by gender (p<0.001). In male patients, an increase of BMI by 1 kg/m2 reduced FEV1 decline by 1.05mL/year (95% CI: 0.96, 1.14). However, in female patients, BMI status barely clinically affected FEV1 decline: an increase of baseline BMI by 1 kg/m2 reduced FEV1 decline by 0.16 ml/year (95% CI: 0.11, 0.21). These gender-modified relationships were generally similar between GOLD 1 and 2 patients, and between current smokers and former smokers.Conclusion: In mild to moderate COPD, higher BMI was associated with a less rapid decline of FEV1 in male patients whereas it hardly affected females patients. This gender-specific BMI effect was independent of COPD severity and smoking status.
Correlation Between Sleep Quality And Quality Of Life Among Hypertensive Adults In A Mountainous Area Of Hubei Province, China
Authors: Ran Li , Chen Xuyu , Li Mengying , Chen Qi , Zhang Yupeng , Tan Xiaodong
Published: July 2020

Background: Hypertension is one of the most common and easy paroxysm diseases. Inadequately controlled hypertension has been related to poor sleep quality, which would be associated with worsening quality of life.Methods: A descriptive analyses was conducted to describe social demographic factors, while ANOVA and t-test were carried out to compare scores between different groups. The total score of life quality (MCS and PCS) was used as the dependent variable (Y), and the dimensions of sleep quality were used as the independent variable (X) for multiple line regression analysis (Stepwise) to evaluate the correlation between sleep quality and life quality.Results: The results of group comparison showed that the total PSQI score was significant at people’s residence (P<0.01). Correlation analysis indicated that subjective sleep quality, sleep disturbance, daytime dysfunction, age, concomitant diseases, and years of diagnosed hypertension had a significant association with the PCS scores (P<0.05 for all). Subjective sleep quality, sleep disturbance, daytime dysfunction, and monthly income had a significant association with the MCS scores (P<0.05 for all). Conclusion: The correlation analysis shows that sleep quality of hypertensive patients is related to quality of life. Considering the close relation among hypertension, sleep quality, and life quality, possible interventions like sleep hygiene was appealed to relieve hypertensive symptoms, promote sleep quality, and increase life quality.Trial registration: 2018-1602000-03-02
Establishment and Histopathological Study of Patient-derived Xenograft Models and Primary Cell Lines of Epithelioid Malignant Peritoneal Mesothelioma
Authors: Yang Zhi-Ran , Chen Zhi-Gao , Ji Zhong-He , Lin Yu-Lin , Zhang Jue , Ma Ru , Li Zhao , Jiang Xi , Chen Qian , Du Xue-Mei , Li Yan
Published: July 2020

Background: Malignant peritoneal mesothelioma (MPM) is a rare malignancy with few effective therapies. This study established patient-derived xenograft (PDX) models and primary cell lines to provide a study platform for MPM in vitro and in vivo, and conducted histopathological analysis.Methods: Human MPM surgical specimen was collected to establish PDX models and primary cell lines. Experimental peritoneal cancer index (ePCI) score was used to evaluate gross pathology, and histopathological study was conducted by hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining. Pathological characteristics of the established primary MPM cell lines were analyzed by Swiss-Gimsa and immunofluorescence (IF) staining. The whole-exome sequencing (WES) was performed to identify the mutant genes between models and the patient.Results: MPM PDX models and primary cell lines were successfully established, replicated the pathological features of the patient. ePCI score of the female and male nude mice were 8.80 ± 1.75 and 9.20 ± 1.81 (P = 0.6219), respectively. The HE staining showed that the tumor was epithelioid mesothelioma and invaded multiple organs. IHC staining showed that Calretinin, Cytokeratin 5/6, WT-1 and Ki-67 were all positive. The Swiss-Gimsa and IF staining of primary cell lines were also consistent with the pathological characteristics of mesothelioma. The WES showed that 21 mutant genes were shared between models and the patient, and the genes related to tumorigenesis and development including BAP1, NF2, MTBP, NECTIN2, CDC23, LRPPRC, TRIM25, and DHRS2.Conclusions: PDX models and primary cell lines of MPM were successfully established with the epithelioid mesothelioma identity confirmed by histopathological evidence. Moreover, our study has also illustrated the shared genomic profile between models and the patient, then potentially provided new targets for MPM treatment.
Evaluation and Application of an Efficient Plant DNA Extraction Protocol in Laboratory and Field Testing
Authors: Wang Qi , Shen Xiaoxia , Qiu Tian , Wu Wei , Wang Zhian , Shou Huixia
Published: July 2020

Background CTAB has been considered as the standard protocol for DNA extraction. But the complex and time-consuming procedures can’t meet the needs of rapid molecular identification. The method of using cellulose filter paper strips to transfer the DNA in the plant tissue lysate to the nucleic acid amplification system shortening the DNA extraction time to 30 s. However, cellulose filter paper strips have some shortcomings that cannot be put into widespread use. And the data supporting the rapidly purification of DNA by cellulose filter paper is not sufficient.ResultsIn the study, the published filter paper strip was modified by sticking the filter paper on the PVC sheet. This modified method is named EZ-D, for easy DNA extraction. Compared with the original method, the DNA extracted by EZ-D is more efficient in PCR amplification. We also came up with a new DNA extraction buffer, which exhibited higher DNA extraction efficiency. When compared with classic CTAB, EZ-D also showed great advantages for higher efficiency, easier protocol and lower cost. PCR analyses showed that DNA extracted from several types of plants by EZ-D were appropriate for specific identification of biological samples. PCR using DNA extracted by EZ-D was sensitive enough to detect 0.1 ng/μL. Evaluation of the EZ-D showed that the DNA extracts can be successfully amplified by PCR reaction for the DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved in an equipment-free way. Conclusion Combined with DNA amplification technology, EZ-D protocol is a rapid, specific and sensitive method for molecular identification of plant samples. In addition, EZ-D method is the first application of cellulose filter paper in the identification of genetically modified crops and traditional Chinese medicine ultra-fine powder. In terms of practicability, EZ-D has realized the popularization of cellulose filter paper for rapid DNA purification in laboratories and markets.
Genome-Wide Methylation Sequencing Identifies Progression-Related Epigenetic Drivers in Myelodysplastic Syndromes
Authors: Zhou Jing-dong , Zhang Ting-juan , Xu Zi-jun , Deng Zhao-qun , Gu Yu , Ma Ji-chun , Wen Xiang-mei , Zhang Wei , Leng Jia-yan , Lin Jiang , Chen Su-ning , Qian Jun
Published: July 2020

Background: The mechanism underlying disease progression of myelodysplastic syndromes (MDS) to acute myeloid leukemia (AML) remains poorly elucidated. Epigenetic alterations are increasingly implicated in the pathogenesis of cancer progression including MDS. However, little studies explored the whole-genome methylation alterations during MDS progression.Methods: We conducted reduced representation bisulfite sequencing in four paired MDS/secondary AML (MDS/sAML) patients and intended to discover methylation-associated epigenetic drivers in MDS progression.Results: In four paired MDS/sAML patients, cases at sAML stage showed significantly increased methylation level when compared with their matched MDS stage. A total of 1090 differentially methylated fragments (DMFs) (441 hypermethylated and 649 hypomethylated) were identified involved in MDS pathogenesis, whereas 103 DMFs (96 hypermethylated and 7 hypomethylated) were identified involved in MDS progression. Further targeted bisulfite sequencing identified aberrant GFRA1, IRX1, NPY, and ZNF300 methylation was frequent events in additional de novo MDS and AML patients, of which only ZNF300 methylation was associated with ZNF300 expression. Moreover, ZNF300 hypermethylation in larger cohorts of de novo MDS and AML patients was further confirmed by real-time quantitative methylation-specific PCR. Clinical studies showed that ZNF300 methylation could act as a potential biomarker helpful for the diagnosis and prognosis in MDS and AML patients. In in vitro experiments, overexpression of ZNF300 exhibited anti-proliferative and pro-apoptotic effects in MDS-derived AML cell line SKM-1.Conclusions: Genome-wide DNA hypermethylation was a frequent event during MDS progression. Among these changes, ZNF300 methylation through the regulation of ZNF300 expression acted as an epigenetic driver in MDS progression. These findings give a theoretical basis for investigating and using the efficacy of DNMT inhibitors in MDS patients against disease progression, and provide new insights for targeted therapy in MDS.
Effects of COVID-19 Outbreak on Children’s Mental Health: A Comparative Study with Children Diagnosed and Isolated from Their Parents
Authors: Kılınçel Şenay , Altun Feray Tarımtay , Nuryüz Özgecan , Tan Ezgi , Erzincan Erkal , Kılınçel Oğuzhan , Yazıcı Esra , Ayaz Muhammed
Published: July 2020

Background
The current study aimed to compare the psychological responses of children exposed to different levels of stress during COVID-19 outbreak. Anxiety levels, negative thoughts, and quality of life of COVID-19 positive children (Child + group, n = 17), COVID-19 negative children who are isolated from their COVID-19 positive parent(s) (Parent + group, n = 59), and a control group (n = 64) were compared.
Methods
The participants completed a sociodemographic data form, the Screen for Child Anxiety and Related Disorders (SCARED), the Children’s Negative Cognitive Errors Questionnaire (CNCEQ), and the Pediatric Quality of Life Inventory (PedsQL).
Results
The statistical analysis of the data obtained from the three groups revealed that the Parent + group recorded higher SCARED and CNCEQ scores and lower PedsQL scores in comparison to the control group. No statistically significant difference was found between the Child + group scores and the control group. It was observed that high CNCEQ scores and low PedsQL scores in the Parent + group predicted an increase in the SCARED scores.
Conclusions
Our study indicates that isolating children from their parents during the COVID-19 outbreak negatively influences their mental health and causes an increase in their anxiety levels. In addition, the results of the study highlight that preventive mental health measures should be applied for children separated from parents during pandemics.
Increase of foreign body and harmful substance ingestion and associated complications in children: a retrospective study of 1199 cases from 2005 to 2017
Authors: Speidel Arne , Woelfle Lena , Mayer Benjamin , Posovszky Carsten
Published: July 2020

Background
Children with a history of caustic or foreign body ingestions (FBI) are frequently presented at the emergency department. An uncertain case history, the different objects and diverse clinical presentation may complicate or delay a proper management and cause serious complications.
Methods
A structured retrospective data analysis of patients presented in between January 2005 and December 2017 at the University Medical Centre Ulm was performed. All patients up to 17 years of age with food impaction, foreign body or harmful substance ingestion were included by selection of the corresponding International Statistical Classification of Diseases and Related Health Problems (ICD10-GM) codes. Descriptive statistics, parametric or non-parametric tests, and linear regression analysis were performed.
Result
In total, 1199 cases were analysed; mean age 3.3 years (SD 3.12; range 7 days to 16 years), male to female ratio 1.15:1, thereof 194 (16.2%) were hospitalized. The majority (n = 619) had no symptoms, 244 out of 580 symptomatic patients complained retching or vomiting. The annually recorded cases increased from 66 in 2005 to 119 in 2017 with a rise of the percentage of all emergency cases by 0.52% since 2010 to 1.34% in 2017. Most frequently, coins were ingested (18,8%). Radiopaque objects accounted for 47.6%, and sharp objects for 10.5% of the FBI, which were significantly more often ingested by girls (p < 0.000015 and p < 0.000024, respectively). Button battery ingestion was recorded in 63 cases with a significant annual increase (R2 = 0.57; beta = 0,753; p < 0.003). The annual rate of complications raised significantly (R2 = 0.42; beta = 0,647; p < 0.017).
Conclusion
Children with FBI are increasingly presented at emergency departments. Special attention is necessary to prevent serious complications, especially in asymptomatic children. Further preventive measures within the home environment are needed to stop this trend.
A Cross-Sectional Study of Olfactory and Taste Disorders in COVID-19 Patients in China
Authors: Li Jianhui , Sun Yi , Li Meirong , Yuan Hu , Yang Changliang , Huang Chengcheng , Zhou Fengjie , Chen Ruiyao , Zhang Leibo , Yu Ning , Liu Qiong , He Jingjing , Zhou Xuejun , Fu Xiaobing , Yang Shiming
Published: July 2020

Objective To determine the prevalance and features of olfactory and taste disorders in coronavirus disease of 2019（CoVID-19）patients in China. Methods A cross-sectional study was performed from 3 April to 15 April 2020 in Wuhan. A total of 187 patients with confirmed severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection completed face-to-face interviews or telephone follow-up. Patients information including epidemiological, clinical, imaging, and serological records and treatment and outcomes data. Patients were further evaluated using questionnaires and visual analogue scale. Statistical analysis was performed with SPSS software (version 22.0). Results The enrolled 187 patients, 23 (12.3%) reported olfactory disorders, 42 (22.46%) reported taste disorders. Females are more prone to olfactory and taste disorders. Among patients that can clearly describe the fluctuation of olfactory and taste disfunctions, 4 (20.0%) and 7(18.92%) cases occurred before other symptoms appeared, the average number of days in advance was 3.5 and 3.57, respectively. The majority of patients can return to normal or basic normal, but in 4(17.39%) and 9(21.43%) cases olfactory and taste can not fully recoveried during the research period. Conclusions Prevalence of olfactory and taste disorders substantially lower in China cohorts compared to abroad COVID-19 cohorts. The prevalence of olfactory and taste disorders in COVID-19 patients was higher in females than in males. In some patients, olfactory and taste disorders precede other symptoms and can be used as a symbol for early screening and warning. The restoration of olfactory and taste function was independent of age; females recover more easily than males; olfactory or taste disorders was not easily recovered for patients with clinically classified as severe; when olfactory or taste disorders itself was serious, it was not easy to recover; olfactory or taste disorders occured early in the disease were more likely to be recovered, otherwise they were hard to be recovered.
Permeability of the Windows of the Brain: Feasibility of Dynamic Contrast-enhanced MRI of the Circumventricular Organs
Authors: Verheggen Inge C.M. , de Jong Joost J.A. , Boxtel Martin P.J. van , Postma Alida A. , Verhey Frans R.J. , Jansen Jacobus F.A. , Backes Walter H.
Published: July 2020

BackgroundCircumventricular organs (CVOs) are small structures without a blood-brain barrier surrounding the brain ventricles that serve homeostasic functions and facilitate communication between the blood, cerebrospinal fluid and brain. Secretory CVOs release peptides and sensory CVOs regulate signal transmission. However, pathogens may enter the brain through the CVOs and trigger neuroinflammation and neurodegeneration. We investigated the feasibility of dynamic contrast-enhanced (DCE) MRI to assess the CVO permeability characteristics in vivo, and expected significant contrast uptake in these regions, due to blood-brain barrier absence.MethodsTwenty healthy, middle-aged to older males underwent brain DCE MRI. Pharmacokinetic modeling was applied to contrast concentration time-courses of CVOs, and in reference to white and gray matter. We investigated whether a significant and positive transfer from blood to brain could be measured in the CVOs, and whether this differed between secretory and sensory CVOs or from normal-appearing brain matter.ResultsIn both the secretory and sensory CVOs, the transfer constants were significantly positive, and all secretory CVOs had significantly higher transfer rates than each sensory CVO. The transfer constants in both the secretory and sensory CVOs were higher than in the white and gray matter.ConclusionsCurrent measurements confirm the often-held assumption of highly permeable CVOs, of which the secretory types have the strongest blood-to-brain transfer. The current study suggests that DCE MRI could be a promising technique to further assess the function of the CVOs and how pathogens can potentially enter the brain via these structures.Trial registrationNetherlands Trial Register number: NL6358, date of registration: 2017-03-24
Camptothecin regulates microglia polarization and exerts neuroprotective effects via the AKT/Nrf2/HO-1-NF-κB signal axis in vivo and in vitro
Authors: he dewei , Liu Dianfeng , Zhou Ang , Gao Xiyu , Zhang Yufei , Su Yingchun , Huang Bingxu , Du Jian , Fu Shoupeng
Published: July 2020

Background

Parkinson's disease (PD), the second largest neurodegenerative disease seriously affects human health. Microglia, the main immune cells in the brain participate in the innate immune response in the central nervous system (CNS). Studies have shown that microglia can be polarized into pro-inflammatory M1 and anti-inflammatory M2 phenotypes. Accumulated evidences suggest that over-activated M1 microglia release pro-inflammatory mediators that damage neurons and lead to Parkinson's disease (PD). In contrast, M2 microglia release neuroprotective factors and exert the effects of neuroprotection. Camptothecin (CPT), an extract of the plant Camptotheca acuminate, has been reported to have anti-inflammation and antitumor effects. However the effect of CPT on microglia polarization and microglia-mediated inflammation responses has not been reported. Therefore, we aim to explore the effect of CPT on microglia polarization and its underlying mechanism on neuroinflammation.
Methods

C57BL/6 mice (25–30 g) were injected LPS or PBS into the substantia nigra (SN). Open-Field Test and Immunohistochemistry were performed to test the dyskinesia of mice and the loss of neurons in the substantia nigra (SN). Microglia cell line BV-2, the neuroblastoma SH-SY5Y and dopaminergic neuron MN9D cell were cultured. Cytotoxicity assay, reverse transcription quantitative real-time polymerase chain reaction (RT-PCR), Western blot, ELISA and Immunofluorescence staining were performed. All results were presented with mean ± SD.
Results

In vivo, CPT improved dyskinesia of mice, reduced the loss of neurons in the substantia nigra (SN) and inhibited neuro-inflammatory responses in LPS-injected mice. In vitro, CPT inhibited M1 polarization of microglia and promotes M2 polarization via the AKT/Nrf2/HO-1-NF-κB signal axis. Furthermore, CPT protected the neuroblastoma cell line SH-SY5Y and dopaminergic neuron cell line MN9D from neurotoxicity of mediated by microglia activation.
Conclusion

CPT regulates the microglia polarization phenotype via the AKT/Nrf2/HO-1-NF-κB signal axis, inhibits neuro-inflammatory responses and exerts neuroprotective effects in vivo and in vitro.
Study on the Demand and Utilization of Health Services for the Poor Elderly in China
Authors: Zeng Yanbing , Xu Weiqian , Luo Bei , Fang Ya
Published: July 2020

Background

Despite historic achievements in fighting poverty over recent decades, the unmet health needs of the poor elderly are still a severe problem in China. This paper investigates the demand and utilization status of health services among the poor elderly and examines the factors that affect healthcare service utilization.
Methods

According to the international poverty line of each year, we selected the poor elderly from the China Health and Retirement Longitudinal Study (CHARLS) from 2011 to 2015 to create nationally representative estimates. The main outcome measures include utilization indicators for the probability and costs of outpatient/inpatient services. Based on a modified Andersen behaviour model, a two-part model is designed to investigate and analyse how predisposing factors, enabling factors, and need and health behaviour variables affect the health service utilization of the poor elderly. The prediction of marginal effect of the unconditional healthcare expenditure are estimated by jointing the model of probability and expenditure of health service utilization.
Results

The morbidity and chronic disease rates of the poor elderly were 16.93% and 79.43%, respectively. The visit rate of outpatient services increased from 15.05% in 2011 to 21.26% in 2015, and the hospitalization rate increased from9.87–16.89%. In terms of medical expenditures, the median cost of outpatient and inpatient services in 2015 for the poor elderly were 350 RMB and 10,000 RMB, respectively, and the out-of-pocket ratios were 85.2% and 53.3% respectively. Overall, 78.44% of the people who actually needed healthcare did not use health services, and the main reason was financial difficulties (42.32%). The two-part model showed that the poor elderly who are higher educated, have children, live in central regions, and have social security and a poor health status and who do not smoke or drink are more likely to use health services. Males who live in the eastern regions with poor self-reported health, physical disabilities and chronic diseases have more medical expenses. Margin effect analysis finds that the need factor is more important than other factors in determining health service utilization.
Conclusion

The poor elderly in China, as the most vulnerable group, tends to have a worse health status and a heavier medical burden. Due to a variety of constraints, the poor elderly had a lower utilization of health services. Predisposing, enabling, need and health behaviour variables should be considered when making policy and taking measures to eliminate health inequalities to improve the health service utilization and the health of the poor elderly.
Characteristics and diversity of mutations in regulatory genes of resistance-nodulation-cell division efflux pumps in association with drug-resistant clinical isolates of Acinetobacter baumannii
Authors: Salehi Bahare , Ghalavand Zohreh , Yadegar Abbas , Eslami Gita
Published: July 2020

Background
This study aimed to characterize the regulation and expression of three putative resistance-nodulation-cell division (RND)-type efflux systems and their contribution to multidrug efflux in clinical isolates of Acinetobacter baumannii.
Methods
Antimicrobial susceptibility testing (AST) of 95 A. baumannii isolates was determined by Kirby-Bauer disk diffusion for 18 antibiotics and minimum inhibitory concentration (MIC) of colistin was determined by broth microdilution method. Moreover, MIC of five classes of antibiotics was assessed using E-test strips in the presence and absence of phenylalanine-arginine beta-naphthylamide (PAβN). Regulatory genes of RND efflux pumps (AdeRS, AdeL, AdeN and BaeSR) were subjected to sequencing. The relative expression of adeB. adeG and adeJ genes was determined by quantitative real-time PCR (RT-PCR).
Results
Overall, majority of isolates (93%) were extensively drug-resistant (XDR). In the phenotypic assay, efflux pump activity was observed in 40% of isolates against multiple antibiotics mainly tigecycline, but not to imipenem. Several amino acid substitutions were detected in the regulatory genes; except in AdeN. Of note, A136V in AdeR and G186V in AdeS were found to be associated with overexpression of their relative efflux pumps. No insertion sequences (ISs) were detected.
Conclusions
Our findings outline the role of RND efflux pumps in resistance of A. baumannii against multiple antibiotics particularly tigecycline, and point out importance of a variety of single mutations in the corresponding regulatory systems. Even though it has been concluded that multidrug resistance occurs as a result of a complex sets of different resistant mechanisms.
Aberrant Methylation Modifications Reflect Specific Drug Responses in Small Cell Lung Cancer
Authors: Chen Peixin , Guo Haoyue , Liu Yu , Chen Bin , Zhao Sha , Wu Shengyu , Li Wei , Wang Lei , Jia Keyi , Wang Hao , Jiang Minlin , Tang Xuzhen , Qi Hui , Dai Chunlei , Ye Junyan , He Yayi
Published: July 2020

Background: Epigenetic alterations of specific genes are closely related to chemotherapeutic efficacy in various cancers. However, few comprehensive profiles of differentially methylated genes (DMGs) in small cell lung cancer (SCLC) were available. The current study aimed to explore the association between aberrant methylation modifications of coding and non-coding genes and chemotherapy insensitivity, thus constructing lncRNA-miRNA-mRNA regulatory network in SCLC.Methods: In this study, methylated DNA immunoprecipitation sequencing (MeDIP-Seq) and RNA sequencing (RNA-Seq) was employed to six SCLC patients who were sensitive and insensitive to first-line standard chemotherapy. Then, we verified the therapeutic predictive effects of DMGs in 62 SCLC cell lines and constructed lncRNA-miRNA-mRNA network on the basis of different cohorts and databases. Results: Of 4552 DMGs which were selected (fold change ≥ 2, p<0.01) between chemo-sensitive and chemo-insensitive group, coding genes constituted the largest percentage (3873/4551, 85.08%), followed by lncRNAs (479/4551, 10.52%) and miRNAs (162/4552, 3.56%). Further, both two groups demonstrated two methylation peaks near transcription start site and transcription end site. Two lncRNA-miRNA-mRNA networks which based on genes that showed opposite results in genetic expression and epigenetic modifications suggested the extensive genome connection between chemotherapy efficacy-related non-coding RNAs (ncRNAs) and mRNAs. Combing four DMGs, including hsa-miR-34a, LINC00461, LINC01018, and LINC01484 could effectively predict first-line chemotherapy response in SCLC, indicating that they might play vital roles in SCLC chemotherapy insensitivity. Conclusions: This study was the first to evaluate multiple drugs efficacy-related ncRNAs and mRNAs which were modified by methylation in SCLC. In addition, DMGs identified in our research might serve as predictors for chemotherapy effect and promising therapeutic targets to reverse drugs-insensitivity by complex lncRNA-miRNA-mRNA mechanisms in SCLC. Further comprehensive studies of these findings are worthy of expecting.
EBV Encoded miRNA BART8-3p Drives Radioresistance-Associated Metastasis in Nasopharyngeal Carcinoma
Authors: Zhou Xiaohan , Lin Yanling , Chen Yuting , Wang Lingzhi , Liao Jinrong , Zeng Hanyi , Luo Wenxiao , Wu Dehua , Cai Longmei
Published: July 2020

Background: Radiotherapy plays an important role in the treatment of nasopharyngeal carcinoma (NPC), however, 20 % of patients with NPC exhibit unusual radioresistance. Patients with radioresistance are at risk of recurrence, so it is imperative to explore the mechanism of resistance to radiotherapy. In the past, studies on the mechanism of radioresistance have been restricted to DNA damage and related cell cycle remodeling or apoptosis. So far, no studies have explored the relationship between radioresistance and metastasis. Methods: We analyzed the metastasis rate of patients with recurrent NPC and that of patients with primary NPC. Constructing an acquired radioresistant NPC cell line and detect their metastatic ability in vivo and in vitro. RNA-deep sequencing was performed to predict the targeted host genes of EBV-miR-BART8-3p. Western blotting, real-time PCR and immunochemistry were conducted to investigate the relationship of clinicopathologic features and EBV-miR-BART8-3p or PAG1. Results:Through the analysis of clinical samples, we observed that the metastasis rate of recurrent NPC was much higher than that of primary patients. In vitro and in vivo experiments showed that NPC cells with acquired radioresistance exhibited a stronger ability for invasion and metastasis. Mechanistically, we found that the Epstein–Barr virus (EBV)-encoded miRNA BART8-3p was increased in patients with NPC and its expression was positively correlated with adverse prognostic factors, such as radioresistance. Besides, miR-BART8-3p promoted the epithelial-mesenchymal transition (EMT), invasion, and metastasis of radioresistant NPC cells by targeting and inhibiting their PAG1 host gene. Conclusion: These results demonstrated a correlation between radioresistance and metastasis in NPC, which depended on the elevated levels of the EBV-encoded miRNA BART8-3p and the inhibition of the PAG1 host gene. These findings suggested a novel role for EBV-miR-BART8-3p and PAG1 in recurrence NPC and highlighted their potential value as prognostic indicators or therapeutic targets.
TMPRSS2, a SARS-CoV-2 Internalization Protease is Downregulated in Head and Neck Cancer Patients.
Authors: Sacconi Andrea , Donzelli Sara , Pulito Claudio , Ferrero Stefano , Morrone Aldo , Rigoni Marta , Pimpinelli Fulvia , Ensoli Fabrizio , Sanguineti Giuseppe , Pellini Raul , Agrawal Nishant , Izumchenko Evgeny , Ciliberto Gennaro , Giannì Aldo , Muti Paola , Strano Sabrina , Blandino Giovanni
Published: July 2020

Background: SARS-coronavirus-2 enters host cells through binding of the Spike protein to ACE2 receptor and subsequent S priming by the TMPRSS2 protease. We aim to assess differences in both ACE2 and TMPRSS2 expression in normal tissues from oral cavity, pharynx, larynx and lung tissues as well as neoplastic tissues from the same areas.Methods: The study has been conducted using the TCGA and the Regina Elena Institute databases and validated by experimental model in HNSCC cells. We also included data from one COVID19 patient who went under surgery for HNSCC.Results: TMPRSS2 expression in HNSCC was significantly reduced compared to the normal tissues. It was more evident in women than in men, in TP53 mutated versus wild TP53 tumors, in HPV negative patients compared to HPV positive counterparts. Functionally, we modeled the multivariate effect of TP53, HPV, and other inherent variables on TMPRSS2. All variables had a statistically significant independent effect on TMPRSS2. In particular, in tumor tissues, HPV negative, TP53 mutated status and elevated TP53-dependent Myc-target genes were associated with low TMPRSS2 expression. The further analysis of both TCGA and our institutional HNSCC datasets identified a signature anti-correlated to TMPRSS2. As proof-of-principle we also validated the anti-correlation between microRNAs and TMPRSS2 expression in a SARS-CoV-2 positive HNSCC patient tissues Finally, we did not find TMPRSS2 promoter methylation.Conclusions: Collectively, these findings suggest that tumoral tissues, herein exemplified by HNSCC and lung cancers might be more resistant to SARS-CoV-2 infection due to reduced expression of TMPRSS2. These observations may help to better assess the frailty of SARS-CoV-2 positive cancer patients.
Characterization of Neoplastic Cells in the Cystic Space of Invasive Micropapillary Carcinoma of the Canine Mammary Gland.
Authors: Rodrigues Michele Angela , Caldeira-Brant Andre L. , Gomes Dawidson Assis , Silveira Tatiany L. , Chiarini-Garcia Hélio , Cassali Geovanni Dantas
Published: July 2020

Background: Invasive micropapillary carcinoma (IMPC) is a rare breast malignant tumor and a variant form of invasive ductal carcinoma that is an aggressive neoplasm of the canine mammary gland and the human breast. There is a progressed recognition of the importance of the tumor microenvironment in cancer development, but little is known about cell types expressed in the cystic space of canine IMPC. This study aimed to investigate the neoplastic and stromal cells surrounding the cystic space in IMPC. Methods: It was used immunohistochemistry (IHC), immunofluorescence (IF), super-resolution microscope and transmission electron microscopy (TEM) to access the localization and morphology of both stromal cells and epithelial cells in canine IMPC cystic areas.Results: Cells surrounding the cystic space in IMPC were positive for the mesenchymal marker's alpha-smooth muscle actin (aSMA), Vimentin, and S100A4. Furthermore, myoepithelial cell marker p63 was negative on IMPC. Tumoral reversal polarity was observed using MUC1 for the first time in IMPC from the canine mammary gland. MUC1 is known to have a role in lumen formation and has an inhibitory role in the cell to stroma interaction. TEM showed that cells lining the IMPC cystic space were modified myoepithelial cells. Conclusion: The present work demonstrates, for the first time, a characterization of the cystic space compound on IMPC from the canine mammary gland. These findings could be useful to understand better the cellular microenvironment in invasive tumors of the mammary gland to improve cancer treatment.
Qualitative Analysis of the Coordination of Major System Change Within the Colombian Health System in Response to COVID-19: Study Protocol
Authors: Turner Simon , Niño Natalia
Published: July 2020

Background
Coronavirus (COVID-19) is posing a major and unprecedented challenge to health service planning and delivery across health systems internationally. This nationally-funded study is analysing the response of the Colombian health system to the COVID-19 pandemic, drawing on qualitative case studies of three local health systems within the country. The approach will be informed by the concept of 'major system change' - or coordinated change among a variety of healthcare organisations and other relevant stakeholders - in order to identify processes that both enable and inhibit adaptation of health services to the challenges presented by COVID-19. The study will collect information on capacity “bottlenecks” as well as successful practices and forms of innovation that have emerged locally, which have the potential for being 'scaled up' across Colombia's health services.
Methods/design
This qualitative study will be undertaken in two phases. In the first, up to 30 stakeholder interviews will be conducted to ascertain and share immediate challenges and opportunities for improvement in response to COVID-19 that can be shared in a timely way with health service leaders to inform immediate health service planning. The stakeholders will be selected in order to sample a range of planning, provider and intermediary organizations within the health system at the national level. In the second, up to 60 further interviews will be conducted to develop in-depth case studies of three local health systems at the metropolitan area level within Colombia. The interview data will be supplemented with documentary analysis and, where feasible, non-participant observation of meetings directly related to the planning and implementation of actions responding to COVID-19. The study will also establish mechanisms for providing timely, formative feedback to inform health system planning relating to COVID-19 and future pandemics by ascertaining stakeholders' preferences during the interviews and wider scoping discussions.
Discussion
The study's findings will aid evaluation of the relevance of the concept of major system change in a context of 'crisis' decision-making and contribute to international lessons on improving health systems' capacity to respond to COVID-19 and future pandemics. Study findings will be shared among various stakeholders in the Colombian healthcare system in a formative and timely way in order to inform healthcare planning in response to COVID-19 and future pandemics. Conducting the study at a time of COVID-19 raises a number of practical issues (including physical distancing and pressure on health services) which have been anticipated in the study design and research team's ways of working.
COVID-19 induced encephalopathy – a Case Report
Authors: Abdelnaby Ramy , Elsayed Mohamed , Abele-Haupts Francis , Barkin Mehmet E. , Rudek Markus A. , Schmidt Kai
Published: July 2020

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also called coronavirus disease 2019 (COVID-19), first appeared in December 2019 in Wuhan, China. It has rapidly spread to multiple countries and has become a global health problem. The effects of COVID-19 on the CNS (Central Nervous System) are reported in low but increasing numbers. We report a case of COVID-19-induced encephalopathy with a biphasic clinical presentation only after a neurologically silent period.
Identification of Key Regulators Responsible for Dysregulated Networks in Osteoarthritis by Large-Scale Expression Analysis
Authors: Shi Song , Wan Fuyin , Zhou Zhenyu , Tao Ran , Lu Yue , Zhou Ming , Liu Fan , Liu Yake
Published: July 2020

Background: Osteoarthritis (OA) is a worldwide musculoskeletal disorder. However, disease-modifying therapies for OA are not available. Here, we aimed to characterize the molecular signatures of OA and to identify novel therapeutic targets and strategies to improve the treatment of OA.Methods: We collected genome-wide transcriptome data performed on 161 OA and 88 normal human cartilage or synovium tissues from 9 independent datasets. Differential gene expression analysis and functional enrichment were performed to identify genes and pathways that were dysregulated in OA. The computational drug repurposing method was used to uncover drugs that could be repurposed to treat OA.Results: We identified several pathways associated with the development of OA, such as extracellular matrix adhesion, inflammation, carbohydrate metabolism, and collagen organization. By protein-protein interaction (PPI) network analysis, we prioritized several hub genes, such as JUN, IL-6, VAMP8, and SOCS3. Moreover, we repurposed several FDA-approved drugs, such as cardiac glycosides, that could be used in the treatment of OA. Conclusions: We proposed that the hub genes we identified will play a role in cartilage homeostasis and could be important diagnostic and therapeutic targets. Drugs such as protein kinase inhibitors and cardiac glycosides could improve the treatment of OA.
Be-careful! Behind the storm of severe COVID-19, could be hidden macrophage activation syndrome. A case report with excellent outcome.
Authors: Gonçalves Rafaela Silva Guimarães , Victor André da Costa , Tavare Ana Carolina Oliveira Cavalcanti , Duarte Angela Luzia Branco Pinto
Published: July 2020

In severe cases of COVID-19, it is important to note that some laboratory signs may alert to the presence of underlying macrophage activation syndrome (MAS), and we ratify that the classic signs of primary MAS are often not present. Here we show a case report of COVID-19 complicated by MAS treated with high doses of methylprednisolone and intravenous Immunoglobulin, with excellent clinical outcome, avoiding orotracheal intubation indeed. The interpretation of laboratory signs leads to early diagnosis and the introduction of effective therapy.
The accuracy of teleradiologists in diagnosing COVID-19 based on a French multicentric emergency cohort
Authors: Nivet Hubert , Crombé Amandine , Schuster Paul , Ayoub Thomas , Pourriol Laurent , Favard Nicolas , Chazot Alban , Alonzo-Lacroix Florian , Youssof Emile , Cheikh Alexandre Ben , Balique Juien , Porta Basile , Petitpierre François , Bouquet Grégoire , Mastier Charles , Bratan Flavie , Bergerot Jean-François , Thomson Vivien , Banaste Nathan , Gorincour Guillaume
Published: July 2020

Objectives: To evaluate the accuracy of diagnoses of COVID-19 based on chest CT as well as inter-observer agreement between teleradiologists during on-call duty and senior radiologists in suspected COVID-19 patients. Materials and Methods: From 03/13/2020 to 04/14/2020, consecutive suspected COVID-19 adult patients who underwent both an RT-PCR test and chest CT from 15 hospitals were included in this prospective study. Chest CTs were immediately interpreted by the on-call teleradiologist and were systematically blind reviewed by a senior radiologist. Readings were categorised using a five-point scale: (1) normal; (2) non-infectious findings; (3) infectious findings but not consistent with COVID-19 infection; (4) consistent with COVID-19 infection; and (5) typical appearance of COVID-19 infection. The diagnostic accuracy of chest CT and inter-observer agreement using the Kappa coefficient were evaluated over the study period.Results: In total, 513 patients were enrolled, of whom 244/513 (47.6%) tested positive for RT-PCR. First readings were scored 4 or 5 in 225/244 (92%) RT-PCR+ patients, and between 1 and 3 in 201/269 (74.7%) RT-PCR- patients. The data were highly consistent (weighted Kappa=0.87) and correlated with RT-PCR (p<0.001, AUC1st-reading=0.89, AUC2nd-reading=0.93). The negative predictive value for scores of 4 or 5 was 0.91–0.92, and the PPV for a score of 5 was 0.89–0.96 at the first and second readings, respectively. Diagnostic accuracy was consistent over the study period, irrespective of a variable prevalence rate.Conclusion: Chest CT demonstrated high diagnostic accuracy with strong inter-observer agreement between on-call teleradiologists with varying degrees of experience and senior radiologists over the study period.
ASSESSMENT OF THE RISK OF SKIN LESIONS ASSOCIATED WITH DEMODEX MITES IN STUDENTS WITH ACNE VULGARIS
Authors: Radko A. , Melekhovets O. , Kharchenko T. , Melekhovets Y.
Published: July 2020

The study of comorbid pathologies that influence the severity of the disease and impair the effectiveness of treatment is carried out to optimize the treatment of the main disease. Materials and methods. The total of 132 medical students was divided into two groups: the 1st group: 56 students who didn`t have acne elements, the 2nd group: 76 students with mild acne vulgaris. At the baseline and after 12 months epithelial tape-test was performed to define Demodex mites presence. The theory of chances was used to assess the impact of acne on the occurrence of Demodex mites. Results. The presence of Demodex mites was observed in 3.57% of students in the 1st group and in 21.05% of students in the 2nd group on repeated tape-test. The probability value (P) among students of the 1st group was P1 = 0.037; among students of the 2nd group – P2 = 0.266. The odds (O) indicator in the 1st group of students without acne elements was O1 = 0.04, in the 2nd group diagnosed with acne – O2 = 0.21. The odds ratio (OR) value in the case/control groups was 7.20. Confidence intervals (95% CI) – (1.582, 32.765): the lower limit of the confidence interval (2.019, + ∞); the upper limit of the confidence interval (-∞, 25680). The significance test p-value was 0.00533. Conclusion. Acne vulgaris is one of the aggravating factor that increases the risk of Demodex mites.