Identification and Characterization of a Second Wolfram Syndrome Gene

Wolfram Syndrome (WFS) is a debilitating autosomal recessive neurodegenerative disorder characterized by juvenile onset insulin dependent diabetes mellitus (DM) and optic atrophy (OA) as well as a number of neurological and endocrine complications that result in early death due to respiratory complications. Previous research has mapped Wolfram syndrome to chromosome 4p16.1 and the disease has been attributed to mutations in the WFS1 gene affecting the WFS1 protein (wolframin), an ER membrane glycoprotein that plays an important role in the unfolded protein response (UPR) and in intracellular Ca2+ homeostasis. An additional locus for WFS on chromosome 4q22-24 was identified by linkage studies of four Jordanian Bedouin families with 16 affected individuals (El-Shanti et al., 2000). In this study, we attempted to identify the causative gene for the second WFS locus and identified a single missense mutation in a novel highly conserved iron-sulfur binding domain gene, CISD2, in the three consanguineous families of Jordanian descent from the El-Shanti et al. (2000) study (Amr et al., 2007). A G→C transversion at nucleotide 109 predicts an amino acid change from glutamic acid to glutamine (E37Q). Although the amino acid is conserved and the mutation is nonsynonymous, the missense mutation was found to disrupt messenger RNA splicing by eliminating exon 2 which results in the introduction of a premature stop codon. CISD2 is expressed in a wide variety of tissues, including those affected in WFS, the brain and pancreas. The CISD2-encoded protein, ERIS (endoplasmic reticulum intermembrane small protein) localizes to the endoplasmic reticulum but does not appear to interact directly with wolframin. Furthermore, lymphoblastoid cells from affected individuals show a significantly greater rise in intracellular calcium when stimulated with thapsigargin, compared with controls, although no difference was observed in resting concentrations of intracellular calcium. To understand the underlying pathogenesis in WFS2 patients, we examined cell death as well as known stress pathways. Cisd2 was knocked down in three cell lines derived from tissues most affected by the disease, namely rat pancreatic insulinoma cells (INS1), mouse neuroblastoma cells (N1E115), and mouse embryonic stem cell like cells (P19) which could be differentiated into neuronal cells. Cisd2 knockdown in INS1 cells shows an increase in apoptotic cell death and in the expression of the apoptotic markers CHOP and BAX, but no increase in the autophagic marker LC3-II. Assessment of the UPR in CISD2 deficient cells shows no activation of the UPR response, while Cisd2 expression in wild-type INS1 and N1E115 cells did not increase under conditions of ER stress. These findings indicate that there is an increase in apoptosis in WFS2 similar to WFS1 but the pathogenesis involves a molecular mechanism that is different than that in WFS1. Investigation of markers of oxidative stress, another major contributor to diabetes and neurodegeneration, show an increase in expression of the antioxidant enzymes Sod1 and Sod2 as well as an increase in global tyrosine nitration in INS1 Cisd2 knockdown cells compared with controls. Cell death in those cells was exacerbated with addition of known oxidative stressors, thapsigargin and paraquat compared with controls. These findings indicate that oxidative stress is a contributor to WFS2 pathogenesis, but it is not clear whether it is the primary causative factor. A recent article implicated ERIS in the BCL-2 associated inhibition of autophagy (Chang et al., 2009) and showed an increase in levels of autophagy in response to starvation in Cisd2 knockdown in a human epithelial carcinoma cell line (H1299). Starvation of INS1 Cisd2 knockdown cells did not elicit a greater autophagic response compared with controls, but did show an increase in expression of Cisd2. P19 Cisd2 knockdown cells that were differentiated into neurons by retinoic acid treatment did not show an inhibition in differentiation markers, but Cisd2 levels returned to levels similar to pre-differentiation wildtype P19 cells, which indicates that Cisd2 is upregulated during neuronal differentiation. In conclusion, the pathogensis of WFS2 can be attributed to apoptotic death of cells in affected tissues, with oxidative stress and not endoplasmic reticulum stress contributing to the development of disease, while ERIS’ relationship with BCL-2-mediated autophagy and neuronal differentiation suggest its important role in cell differentiation and survival

The Translational Genomics Core at Partners Personalized Medicine: Facilitating the Transition of Research towards Personalized Medicine

The Translational Genomics Core (TGC) at Partners Personalized Medicine (PPM) serves as a fee-for-service core laboratory for Partners Healthcare researchers, providing access to technology platforms and analysis pipelines for genomic, transcriptomic, and epigenomic research projects. The interaction of the TGC with various components of PPM provides it with a unique infrastructure that allows for greater IT and bioinformatics opportunities, such as sample tracking and data analysis. The following article describes some of the unique opportunities available to an academic research core operating within PPM, such the ability to develop analysis pipelines with a dedicated bioinformatics team and maintain a flexible Laboratory Information Management System (LIMS) with the support of an internal IT team, as well as the operational challenges encountered to respond to emerging technologies, diverse investigator needs, and high staff turnover. In addition, the implementation and operational role of the TGC in the Partners Biobank genotyping project of over 25,000 samples is presented as an example of core activities working with other components of PPM

Bond Characteristics of ASTM A1035 Steel Reinforcing Bars

The results of a coordinated research program on the bond characteristics of the high-strength steel reinforcing bars that conform to ASTM A1035 are presented. Concrete with nominal strengths of 5000 and 8000 psi (35 and 55 MPa) were used. Sixtynine large-scale beam-splice specimens were tested. Maximum bar stresses are compared with predictions obtained using the bond equations in the ACI 318-05 code provisions and those proposed by ACI Committee 408. Maximum stress levels of 120, 110, and 96 ksi (830, 760, and 660 MPa) were developed in No. 5, No. 8, and No. 11 (No. 16, No. 25, and No. 36) bars, respectively, not confined by transverse reinforcement. Providing confinement for No. 8 and No. 11 (No. 25 and No. 36) spliced bars using transverse reinforcement allowed stresses of up to 150 ksi (1035 MPa) to be developed. The ACI Committee 408 equation provides a reasonable estimate of the strength for both unconfined and confined splices using a strength reduction factor (f-factor) of 0.82 and design parameters (cover, spacing, and concrete strengths) comparable to those used in this test program. The design equations in ACI 318 are less conservative, with a large percentage of the developed/calculated strength ratios below 1.0, and should not be used for development and splice design with high-strength reinforcing steel in their present form

Medical doctors\u27 offline computer-assisted digital education: Systematic review by the digital health education collaboration

Background: The widening gap between innovations in the medical field and the dissemination of such information to doctors may affect the quality of care. Offline computer-based digital education (OCDE) may be a potential solution to overcoming the geographical, financial, and temporal obstacles faced by doctors.Objective: The objectives of this systematic review were to evaluate the effectiveness of OCDE compared with face-to-face learning, no intervention, or other types of digital learning for improving medical doctors\u27 knowledge, cognitive skills, and patient-related outcomes. Secondary objectives were to assess the cost-effectiveness (CE) of OCDE and any adverse effects.Methods: We searched major bibliographic databases from 1990 to August 2017 to identify relevant articles and followed the Cochrane methodology for systematic reviews of intervention.Results: Overall, 27 randomized controlled trials (RCTs), 1 cluster RCT (cRCT), and 1 quasi-RCT were included in this review. The total number of participants was 1690 in addition to the cRCT, which included 24 practices. Due to the heterogeneity of the participants, interventions, and outcomes, meta-analysis was not feasible, and the results were presented as narrative summary. Compared with face-to-face learning, the effect of OCDE on knowledge gain is uncertain (ratio of the means [RM] range 0.95-1.17; 8 studies, 495 participants; very low grade of evidence). From the same comparison, the effect of OCDE on cognitive skill gain is uncertain (RM range 0.1-0.9; 8 studies, 375 participants; very low grade of evidence). OCDE may have little or no effect on patients\u27 outcome compared with face-to-face education (2 studies, 62 participants; low grade of evidence). Compared with no intervention, OCDE may improve knowledge gain (RM range 1.36-0.98; 4 studies, 401 participants; low grade of evidence). From the same comparison, the effect of OCDE on cognitive skill gain is uncertain (RM range 1.1-1.15; 4 trials, 495 participants; very low grade of evidence). One cRCT, involving 24 practices, investigated patients\u27 outcome in this comparison and showed no difference between the 2 groups with low-grade evidence. Compared with text-based learning, the effect of OCDE on cognitive skills gain is uncertain (RM range 0.91-1.46; 3 trials with 4 interventions; 68 participants; very low-grade evidence). No study in this comparison investigated knowledge gain or patients\u27 outcomes. One study assessed the CE and showed that OCDE was cost-effective when compared with face-to-face learning if the cost is less than or equal to Can $200. No trial evaluated the adverse effect of OCDE.Conclusions: The effect of OCDE compared with other methods of education on medical doctors\u27 knowledge and cognitive skill gain is uncertain. OCDE may improve doctors\u27 knowledge compared with no intervention but its effect on doctors\u27 cognitive skills is uncertain. OCDE may have little or no effect in improving patients\u27 outcome

Empirical comparison of reduced representation bisulfite sequencing and Infinium BeadChip reproducibility and coverage of DNA methylation in humans

We empirically examined the strengths and weaknesses of two human genome-wide DNA methylation platforms: rapid multiplexed reduced representation bisulfite sequencing and Illumina’s Infinium BeadChip. Rapid multiplexed reduced representation bisulfite sequencing required less input DNA, offered more flexibility in coverage, and interrogated more CpG loci at a higher regional density. The Infinium covered slightly more protein coding, cancer-associated and mitochondrial-related genes, both platforms covered all known imprinting clusters, and rapid multiplexed reduced representation bisulfite sequencing covered more microRNA genes than the HumanMethylation450, but fewer than the MethylationEPIC. Rapid multiplexed reduced representation bisulfite sequencing did not always interrogate exactly the same CpG loci, but genomic tiling improved overlap between different libraries. Reproducibility of rapid multiplexed reduced representation bisulfite sequencing and concordance between the platforms increased with CpG density. Only rapid multiplexed reduced representation bisulfite sequencing could genotype samples and measure allele-specific methylation, and we confirmed that Infinium measurements are influenced by nearby single-nucleotide polymorphisms. The respective strengths and weaknesses of these two genome-wide DNA methylation platforms need to be considered when conducting human epigenetic studies

Synthesis of Gemini cationic surfactants based on natural nicotinic acid and evaluation of their inhibition performance at C-steel/1 M HCl interface: Electrochemical and computational investigations

Herein, we prepare effective Gemini cationic surfactants (CSII, CSIV) and characterize them using FT-IR and 1HNMR spectroscopy. The adsorptive properties of CSII and CSIV at HCl/air and C-steel/HCl interfaces were examined with surface tension and electrochemical parameters, respectively. The critical micelle concentration (CMC) of the CSII and CSIV indicated their adsorption affinity at the HCl/air interface. Where, aliphatic chains increase surface coverage percentage and aid in surfactant adsorption. The electrochemical parameters of C-steel in 1 M HCl were studied using electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization (PDP) at different temperatures. The charge transfer resistance of the C-steel electrode was enhanced from 28.2 Ω.cm2 to 770.79 and 831.45 Ω.cm2 after adding 5 × 10−4 M of CSII and CSIV, respectively. Both CSII and CSIV act as mixed inhibitors with inhibition performance exceeding 97% due to their highly adsorption affinity. The chemical adsorption affinity of these compounds is suggested by the higher adsorption energy (∆G*ads) values (>−40 kJ mol−1) according to the Langmuir isotherm model. The theoretical calculations including DFT, and Monte Carlo simulation (MCs) provide insight into the relationship between corrosion inhibition and molecular structure, where the calculated parameters agree with the experimental results

Consensus interpretation of the p.Met34Thr and p.Val37Ile variants in GJB2 by the ClinGen Hearing Loss Expert Panel

Purpose: Pathogenic variants in GJB2 are the most common cause of autosomal recessive sensorineural hearing loss. The classification of c.101T>C/p.Met34Thr and c.109G>A/p.Val37Ile in GJB2 are controversial. Therefore, an expert consensus is required for the interpretation of these two variants. Methods: The ClinGen Hearing Loss Expert Panel collected published data and shared unpublished information from contributing laboratories and clinics regarding the two variants. Functional, computational, allelic, and segregation data were also obtained. Case-control statistical analyses were performed. Results: The panel reviewed the synthesized information, and classified the p.Met34Thr and p.Val37Ile variants utilizing professional variant interpretation guidelines and professional judgment. We found that p.Met34Thr and p.Val37Ile are significantly overrepresented in hearing loss patients, compared with population controls. Individuals homozygous or compound heterozygous for p.Met34Thr or p.Val37Ile typically manifest mild to moderate hearing loss. Several other types of evidence also support pathogenic roles for these two variants. Conclusion: Resolving controversies in variant classification requires coordinated effort among a panel of international multi-institutional experts to share data, standardize classification guidelines, review evidence, and reach a consensus. We concluded that p.Met34Thr and p.Val37Ile variants in GJB2 are pathogenic for autosomal recessive nonsyndromic hearing loss with variable expressivity and incomplete penetrance

Psychological impact of COVID-19 on healthcare workers: cross-sectional analyses from 14 countries

Abstract Background Healthcare workers (HCWs) have been impacted psychologically due to their professional responsibilities over the prolonged era of the coronavirus disease 2019 (COVID-19) pandemic. The study aimed to identify the predictors of psychological distress, fear, and coping during the COVID-19 pandemic among HCWs. Methods A cross-sectional online survey was conducted among self-identified HCWs across 14 countries (12 from Asia and two from Africa). The Kessler Psychological Distress Scale, the Fear of COVID-19 Scale, and the Brief Resilient Coping Scale were used to assess the psychological distress, fear, and coping of HCWs, respectively. Results A total of 2447 HCWs participated; 36% were doctors, and 42% were nurses, with a mean age of 36 (±12) years, and 70% were females. Moderate to very-high psychological distress was prevalent in 67% of the HCWs; the lowest rate was reported in the United Arab Emirates (1%) and the highest in Indonesia (16%). The prevalence of high levels of fear was 20%; the lowest rate was reported in Libya (9%) and the highest in Egypt (32%). The prevalence of medium-to-high resilient coping was 63%; the lowest rate was reported in Libya (28%) and the highest in Syria (76%). Conclusion COVID-19 has augmented the psychological distress among HCWs. Factors identified in this study should be considered in managing the wellbeing of HCWs, who had been serving as the frontline drivers in managing the crisis successfully across all participating countries. Furthermore, interventions to address their psychological distress should be considered

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

SARS-CoV-2 susceptibility and COVID-19 disease severity are associated with genetic variants affecting gene expression in a variety of tissues

Variability in SARS-CoV-2 susceptibility and COVID-19 disease severity between individuals is partly due to genetic factors. Here, we identify 4 genomic loci with suggestive associations for SARS-CoV-2 susceptibility and 19 for COVID-19 disease severity. Four of these 23 loci likely have an ethnicity-specific component. Genome-wide association study (GWAS) signals in 11 loci colocalize with expression quantitative trait loci (eQTLs) associated with the expression of 20 genes in 62 tissues/cell types (range: 1:43 tissues/gene), including lung, brain, heart, muscle, and skin as well as the digestive system and immune system. We perform genetic fine mapping to compute 99% credible SNP sets, which identify 10 GWAS loci that have eight or fewer SNPs in the credible set, including three loci with one single likely causal SNP. Our study suggests that the diverse symptoms and disease severity of COVID-19 observed between individuals is associated with variants across the genome, affecting gene expression levels in a wide variety of tissue types