Work, Parenting, and Well-being: An International Survey of Neurologists During the COVID-19 Pandemic.

Background and Objectives: COVID-19 reframed the relationship between work and home and, in general, made both more difficult-especially for parents. We hypothesized that, among neurologists, the effects of the pandemic on productivity and on well-being would be greater on those with children than on those without children and that the effects would be greater on women with children than on men with children. Methods: We conducted an international electronic survey launched by the Practice Current section of the American Academy of Neurology. The survey included questions on demographics (self-identified gender, number of children and elderly dependents, childcare support, and country and state when applicable), workflow changes because of COVID-19, impacted domains, and productivity and well-being using the Likert scale. Counts are presented as descriptive statistics. Statistical analysis was performed using Mann-Whitney U and Kruskal-Wallis tests. Results: We collected 243 fully completed surveys from providers in all continents with high representation of the United States (76%), providers who identified as women (71.6%), and neurologists with children (91%) among respondents. A majority worked remotely (28% fully, 43% mix). Neurologists reported decreased academic productivity (72%), work benefits (65%), and time for writing (48%). These findings were more prominent in respondents with children and among women practicing outside of the United States. Increased pressure from productivity expectations and lack of time for family were reported by 47% and 41% of respondents, respectively. Discussion: The disruption from the COVID-19 pandemic affected academic productivity and decreased the well-being of neurologists in general and of neurologists with children more drastically. This could potentially hinder the promotion and retention of junior neurologists who were juggling life and work during the pandemic outbreak and its recurrent surges.

Computational identification of a multi-peptide vaccine candidate in E2 glycoprotein against diverse Hepatitis C virus genotypes.

Hepatitis C Virus (HCV) is estimated to affect nearly 180 million people worldwide, culminating in ∼0.7 million yearly casualties. However, a safe vaccine against HCV is not yet available. This study endeavored to identify a multi-genotypic, multi-epitopic, safe, and globally competent HCV vaccine candidate. We employed a consensus epitope prediction strategy to identify multi-epitopic peptides in all known envelope glycoprotein (E2) sequences, belonging to diverse HCV genotypes. The obtained peptides were screened for toxicity, allergenicity, autoimmunity and antigenicity, resulting in two favorable peptides viz., P2 (VYCFTPSPVVVG) and P3 (YRLWHYPCTV). Evolutionary conservation analysis indicated that P2 and P3 are highly conserved, supporting their use as part of a designed multi-genotypic vaccine. Population coverage analysis revealed that P2 and P3 are likely to be presented by >89% Human Leukocyte Antigen (HLA) molecules from six geographical regions. Indeed, molecular docking predicted the physical binding of P2 and P3 to various representative HLAs. We designed a vaccine construct using these peptides and assessed its binding to toll-like receptor 4 (TLR-4) by molecular docking and simulation. Subsequent analysis by energy-based and machine learning tools predicted high binding affinity and pinpointed the key binding residues (i.e. hotspots) in P2 and P3. Also, a favorable immunogenic profile of the construct was predicted by immune simulations. We encourage the scientific community to validate our vaccine construct in vitro and in vivo.Communicated by Ramaswamy H. Sarma.

A discovery-based proteomics approach identifies protein disulphide isomerase (PDIA1) as a biomarker of ß cell stress in type 1 diabetes.

BACKGROUND: Stress responses within the ß cell have been linked with both increased ß cell death and accelerated immune activation in type 1 diabetes (T1D). At present, information on the timing and scope of these responses as well as disease-related changes in islet ß cell protein expression during T1D development is lacking. METHODS: Data independent acquisition-mass spectrometry was performed on islets collected longitudinally from NOD mice and NOD-SCID mice rendered diabetic through T cell adoptive transfer. FINDINGS: In islets collected from female NOD mice at 10, 12, and 14 weeks of age, we found a time-restricted upregulation of proteins involved in stress mitigation and maintenance of ß cell function, followed by loss of expression of protective proteins that heralded diabetes onset. EIF2 signalling and the unfolded protein response, mTOR signalling, mitochondrial function, and oxidative phosphorylation were commonly modulated pathways in both NOD mice and NOD-SCID mice rendered acutely diabetic by T cell adoptive transfer. Protein disulphide isomerase A1 (PDIA1) was upregulated in NOD islets and pancreatic sections from human organ donors with autoantibody positivity or T1D. Moreover, PDIA1 plasma levels were increased in pre-diabetic NOD mice and in the serum of children with recent-onset T1D compared to non-diabetic controls. INTERPRETATION: We identified a core set of modulated pathways across distinct mouse models of T1D and identified PDIA1 as a potential human biomarker of ß cell stress in T1D. FUNDING: NIH (R01DK093954, DK127308, U01DK127786, UC4DK104166, R01DK060581, R01GM118470, and 5T32DK101001-09). VA Merit Award I01BX001733. JDRF (2-SRA-2019-834-S-B, 2-SRA-2018-493-A-B, 3-PDF-20016-199-A-N, 5-CDA-2022-1176-A-N, and 3-PDF-2017-385-A-N).

Roles of Chromatin Remodelling and Molecular Heterogeneity in Therapy Resistance in Glioblastoma.

Cancer stem cells (CSCs) represent a therapy-resistant reservoir in glioblastoma (GBM). It is now becoming clear that epigenetic and chromatin remodelling programs link the stemlike behaviour of CSCs to their treatment resistance. New evidence indicates that the epigenome of GBM cells is shaped by intrinsic and extrinsic factors, including their genetic makeup, their interactions and communication with other neoplastic and non-neoplastic cells, including immune cells, and their metabolic niche. In this review, we explore how all these factors contribute to epigenomic heterogeneity in a tumour and the selection of therapy-resistant cells. Lastly, we discuss current and emerging experimental platforms aimed at precisely understanding the epigenetic mechanisms of therapy resistance that ultimately lead to tumour relapse. Given the growing arsenal of drugs that target epigenetic enzymes, our review addresses promising preclinical and clinical applications of epidrugs to treat GBM, and possible mechanisms of resistance that need to be overcome.

Creating a New Set of Milestones for the Clinical Neurophysiology Fellowship.

INTRODUCTION: The Accreditation Council for Graduate Medical Education and the American Board of Psychiatry and Neurology first developed milestones for the clinical neurophysiology (CNP) fellowship in 2015. The milestones provide a comprehensive evaluation of the fellow's development based on six domains of competency. Here, we describe the development of a new set of milestones for CNP fellowship with level 1 as the incoming level, level 4 as the goal for graduation, and level 5 as the aspirational level that may not be achieved. METHODS: Committee members were nominated or volunteered to participate in the milestones update. Milestone development began with the creation of a shared mental model of the ideal skills and knowledge a graduating CNP fellow should attain. RESULTS: The CNP committee met virtually 7 times for a total of 14 meeting hours. Nine Patient Care and five Medical Knowledge milestones evolved from the seven Patient Care and six Medical Knowledge milestones that were in the first iteration. The committee incorporated 11 "Harmonized Milestones" into the revision and a supplemental guide was created. CONCLUSIONS: The revised Accreditation Council for Graduate Medical Education milestones for CNP fellowship contain important updates that program directors should review against their curricula to identify any gaps in learning. Program leadership should take note of two new Patient Care milestones for telemedicine and intraoperative monitoring. Clinical neurophysiology fellowships are not designed to provide level 4 competency across all milestones. The revised milestones should be viewed within the context of an individual program's goals.

Inhibitory activities of grape bioactive compounds against enzymes linked with human diseases.

A quest for identification of novel, safe and efficient natural compounds, as additives in the modern food and cosmetic industries, has been prompted by concerns about toxicity and side effects of synthetic products. Plant phenolic compounds are one of the most documented natural products due to their multifarious biological applications. Grape (Vitis vinifera) is an important source of phenolic compounds such as phenolic acids, tannins, quinones, coumarins and, most importantly, flavonoids/flavones. This review crisply encapsulates enzyme inhibitory activities of various grape polyphenols towards different key human-ailment-associated enzymes: xanthine oxidase (gout), tyrosinase (hyperpigmentation), α-amylase and α-glucosidase (diabetes mellitus), pancreatic lipase (obesity), cholinesterase (Alzheimer's disease), angiotensin i-converting enzymes (hypertension), α-synuclein (Parkinson's disease) and histone deacetylase (various diseases). The review also depicts the enzyme inhibitory mechanism of various grape polyphenols and briefly discusses their stature as potential therapeutic and drug development candidates. KEY POINTS: • Nineteen major bioactive polyphenols from the grape/grape products and their disease targets are presented • Sixty-two important polyphenols as enzyme inhibitors from grape/grape products are presented • A thorough description and graphical presentation of biological significance of polyphenols against various diseases.

Copy-scAT: Deconvoluting single-cell chromatin accessibility of genetic subclones in cancer.

Single-cell epigenomic assays have tremendous potential to illuminate mechanisms of transcriptional control in functionally diverse cancer cell populations. However, application of these techniques to clinical tumor specimens has been hampered by the current inability to distinguish malignant from nonmalignant cells, which potently confounds data analysis and interpretation. Here, we describe Copy-scAT, an R package that uses single-cell epigenomic data to infer copy number variants (CNVs) that define cancer cells. Copy-scAT enables studies of subclonal chromatin dynamics in complex tumors like glioblastoma. By deploying Copy-scAT, we uncovered potent influences of genetics on chromatin accessibility profiles in individual subclones. Consequently, some genetic subclones were predisposed to acquire stem-like or more differentiated molecular phenotypes, reminiscent of developmental paradigms. Copy-scAT is ideal for studies of the relationships between genetics and epigenetics in malignancies with high levels of intratumoral heterogeneity and to investigate how cancer cells interface with their microenvironment.

Current Status and Future Strategies for Mentoring Women in Neurology.

The American Academy of Neurology's (AAN) 2017 Gender Disparity Report identified improving mentorship as a key intervention to fill the leadership and pay gaps for women in neurology. Here we summarize the literature on mentoring women, provide an outline of ideal components of programs geared toward closing gender gaps, and present a mentoring program for AAN members. The strategies discussed share similarities with those for closing gaps related to race, ethnicity, and religion. Developing effective mentorship and sponsorship programs is essential to ensure a sufficiently diverse pool of academic faculty and private practitioners and to establish equal representation in leadership roles in this field.

Health anxiety, coping mechanisms and COVID 19: An Indian community sample at week 1 of lockdown.

It is critical to gain an understanding of the impact of the COVID 19 pandemic and the associated lockdown restrictions on the psychological, social and behavioural functioning of the general public, in order to inform public health promotion and future health service resource allocation. This cross-sectional study, completed during week 1 of lockdown in India, reports on data from 234 participants using an online survey. Data regarding health anxiety, coping mechanisms and locus of control was collected. Structural equation modelling was used to assess the relationship between locus of control, coping mechanisms, health anxiety and age. Age related differences in both locus of control and coping strategies were found. Younger people experienced more health-related anxiety and were more likely to engage with social media as a coping mechanism. Mindfulness-based strategies may reduce health anxiety by increasing tolerance of uncertainty experienced during the COVID 19 pandemic.

Top Ten Tips Palliative Care Clinicians Should Know About Seizures at the End of Life.

Seizures are physically burdensome and emotionally distressing for patients, families, caregivers, and clinicians. Patients with neurological diseases are at increased risk of having complex, difficult-to-control seizures at the end of life. Palliative care (PC) clinicians asked to provide management of these seizures may not be familiar or comfortable with more complex seizures or epilepsy. A team of neurologists and PC specialists have compiled a list of tips to guide clinicians on how to care for patients having seizures and to support their families/caregivers.

Single-cell chromatin accessibility profiling of glioblastoma identifies an invasive cancer stem cell population associated with lower survival.

Chromatin accessibility discriminates stem from mature cell populations, enabling the identification of primitive stem-like cells in primary tumors, such as glioblastoma (GBM) where self-renewing cells driving cancer progression and recurrence are prime targets for therapeutic intervention. We show, using single-cell chromatin accessibility, that primary human GBMs harbor a heterogeneous self-renewing population whose diversity is captured in patient-derived glioblastoma stem cells (GSCs). In-depth characterization of chromatin accessibility in GSCs identifies three GSC states: Reactive, Constructive, and Invasive, each governed by uniquely essential transcription factors and present within GBMs in varying proportions. Orthotopic xenografts reveal that GSC states associate with survival, and identify an invasive GSC signature predictive of low patient survival, in line with the higher invasive properties of Invasive state GSCs compared to Reactive and Constructive GSCs as shown by in vitro and in vivo assays. Our chromatin-driven characterization of GSC states improves prognostic precision and identifies dependencies to guide combination therapies.

Representation of women plenary speakers at the American Academy of Neurology Annual Meeting.

OBJECTIVE: To determine whether women have been equitably represented among plenary speakers at the American Academy of Neurology (AAN) Annual Meeting by counting and categorizing speakers and comparing outcomes to AAN membership and US neurology workforce data. METHODS: Lists of plenary speakers between 1958 and 2019 (62 years) were obtained from the AAN. The primary outcome measures were numbers and proportions of men and women in aggregate and among physicians. RESULTS: We identified 635 plenary speakers, including 148 (23.3%) women. Specifically, women made up 14.6% (19 of 130) of presidential and 25.5% (129 of 505) of nonpresidential plenary session speakers. The inclusion of women plenary speakers was meaningfully higher (h = 0.33; difference 14.9%; 95% confidence interval 4.2%-26.7%) for nonphysicians (27 of 74 [36.5%]) than physicians (121 of 561 [21.6%]). Although at zero levels for Annual Meetings held between 1958 and 1990 and at mostly low but varying levels thereafter, the representation of women and women physicians has been at or above their proportions in the AAN membership and US neurology workforce since 2017. Comparison of representation by plenary session name revealed an unequal distribution of women, with women physicians concentrated in the Sidney Carter Award in Child Neurology presidential session. CONCLUSION: Historically and recently, women and women physicians were underrepresented among AAN plenary speakers. As the AAN has taken active steps to address equity, women have been included in more representative proportions overall. However, notable gaps remain, especially in specific prestigious plenary sessions, and further research is needed to determine causality.

Differences in treatment of epilepsy in pregnancy: A worldwide survey.

BACKGROUND: How to safely treat pregnant women with epilepsy is a question for which there are guidelines, but variations in practice exist. METHODS: To better characterize how clinicians address this difficult clinical question, we distributed an anonymous survey to neurology practitioners across subspecialties and different levels of training via the Neurology®: Clinical Practice website. The survey was conducted from May 31 to December 3, 2017. We received responses from 642 participants representing 81 countries. We performed both descriptive and inferential analyses. For the inferential analysis, a multiple logistic regression model was used to analyze the effect of provider characteristics on the constructed binary outcome variables of interest. RESULTS: The results of this survey demonstrate a wide range in the amount of folic acid recommended and the frequency of checking levels of anti-epileptic drugs. Choice of first-line agent varied by the economic development status of the respondent's country, suggesting that access to medications plays an important role in clinical decision making in many parts of the world. CONCLUSION: This survey highlights several areas where further research would be helpful in guiding practice.

Turn-On Fluorescent Sensors for the Selective Detection of Al3+ (and Ga3+) and PPi Ions.

Rationally designed multiple hydroxyl-group-based chemosensors L1-L4 containing arene-based fluorophores are presented for the selective detection of Al3+ and Ga3+ ions. Changes in the absorption and emission spectra of L1-L4 in ethanol were easily observable upon the addition of Al3+ and Ga3+ ions. Competitive binding studies, detection limits, and binding constants illustrate significant sensing abilities of these chemosensors with L4, showing the best results. The interaction of Al3+/Ga3+ ions with chemosensor L4 was investigated by fluorescence lifetime measurements, whereas Job's plot, high-resolution mass spectrometry, and 1H NMR spectral titrations substantiated the stoichiometry between L4 and Al3+/Ga3+ ions. The solution-generated [L-M3+] species further detected pyrophosphate ion (PPi) by exhibiting emission enhancement and a visible color change. The binding of Al3+/Ga3+ ions with chemosensor L4 was further supported by density functional theory studies. Reversibility for the detection of Al3+/Ga3+ ions was achieved by utilizing a suitable proton source. The multiionic response, reversibility, and optical visualization of the present chemosensors make them ideal for practical applications for real samples, which have been illustrated by paper-strip as well as polystyrene film-based detection.

Burnout and Career Satisfaction in Women Neurologists in the United States.

BACKGROUND: Professional burnout is a growing problem among physicians. Neurology has been found to be one of the specialties with the highest prevalence for burnout. However, little is known about gender-specific risk factors. The objectives of this study were (1) to determine the prevalence of burnout among a sample of women neurologists in the United States and (2) to identify predictive factors leading to burnout. MATERIALS AND METHODS: An online survey was distributed to 798 U.S. women neurologists through the closed Facebook group Women Neurologists Group. Burnout was assessed with the Mini-Z survey. Additional questions assessed current practice settings, family and childcare responsibilities, work-life balance, gender discrimination experiences, career satisfaction, and plans for career changes. RESULTS: The survey received 181 responses, yielding a 22.7% response rate. Most respondents were 1-10 years post-training and 35.4% indicated they felt neutral or dissatisfied toward their current job; 42.6% of respondents reported symptoms of burnout. Working in a high stress environment, lack of control over the work schedule, a higher number of hours at work, and self-reported gender discrimination were each independent predictive factors for burnout. Having more children was associated with decreased likelihood of becoming a physician again, and less than a third of respondents with three or more children indicated they would become a physician again. While 91.1% of respondents considered themselves effective with electronic health record use, 56.9% indicated insufficient time for documentation. CONCLUSIONS: Professional burnout and career dissatisfaction have high prevalence in women neurologists and threaten the future of the neurology workforce. There is an urgent need for interventions to alleviate stressors associated with burnout and measures to reduce gender discrimination.

Age and sex differences in burnout, career satisfaction, and well-being in US neurologists.

OBJECTIVE: To examine age and sex differences in burnout, career satisfaction, and well-being in US neurologists. METHODS: Quantitative and qualitative analyses of men's (n = 1,091) and women's (n = 580) responses to a 2016 survey of US neurologists. RESULTS: Emotional exhaustion in neurologists initially increased with age, then started to decrease as neurologists got older. Depersonalization decreased as neurologists got older. Fatigue and overall quality of life in neurologists initially worsened with age, then started to improve as neurologists got older. More women (64.6%) than men (57.8%) met burnout criteria on univariate analysis. Women respondents were younger and more likely to work in academic and employed positions. Sex was not an independent predictive factor of burnout, fatigue, or overall quality of life after controlling for age. In both men and women, greater autonomy, meaning in work, reasonable amount of clerical tasks, and having effective support staff were associated with lower burnout risk. More hours worked, more nights on call, higher outpatient volume, and higher percent of time in clinical practice were associated with higher burnout risk. For women, greater number of weekends doing hospital rounds was associated with higher burnout risk. Women neurologists made proportionately more negative comments than men regarding workload, work-life balance, leadership and deterioration of professionalism, and demands of productivity eroding the academic mission. CONCLUSIONS: We identified differences in burnout, career satisfaction, and well-being in neurologists by age and sex. This may aid in developing strategies to prevent and mitigate burnout and promote professional fulfillment for different demographic subgroups of neurologists.

Distribution and Quantity of Sites of John Cunningham Virus Persistence in Immunologically Healthy Patients: Correlation With John Cunningham Virus Antibody and Urine John Cunningham Virus DNA.

Importance: Although seroepidemiological studies indicate that greater than 50% of the population has been infected with John Cunningham virus (JCV), the sites of JCV persistence remain incompletely characterized. Objective: To determine sites of JCV persistence in immunologically healthy individuals. Design, Setting, and Participants: Tissue specimens from multiple sites including brain, renal, and nonrenal tissues were obtained at autopsy performed in the Department of Pathology at the University of Kentucky from 12 immunologically healthy patients between February 9, 2011, and November 27, 2012. Quantitative polymerase chain reaction was performed on the tissue specimens and urine. Serum JCV antibody status was determined by enzyme-linked immunosorbent assay. Main Outcomes and Measures: The detection and quantification of JCV from the tissues by quantitative polymerase chain reaction illuminated sites of viral persistence. These results were correlated with JCV antibody levels. Results: Autopsies were performed on 12 individuals, 10 men and 2 women, ranging in age from 25 to 75 years (mean, 55.3 years). Seven of 12 individuals were JCV antibody seropositive based on absorbance units. Serostatus was associated with amounts of JCV DNA in urine and its tissue distribution. John Cunningham virus DNA was found in 75% of genitourinary tissue samples from donors (18 of 24) with high JCV antibody levels, 13.3% of donors with low levels i(4 of 30), and 0% of seronegative persons (0 of 32). In nongenitourinary tissues, JCV DNA was detected in 45.1% of tissue samples of donors (32 of 71) with high JCV, 2.2% of donors with low JCV serostatus (2 of 93), and 0% of seronegative persons (0 of 43). Genitourinary tissues had higher copy numbers than other sites. John Cunningham virus DNA was detected in urine of seronegative individuals in a research-grade assay. Conclusions and Relevance: Persistent (latent or actively replicating) JCV infection mostly predominates in genitourinary tissues but distributes in other tissues at low copy number. The distribution and copy numbers of the virus appear to correlate with urinary JCV shedding and serostatus.

A highly selective chromogenic sensor for Mn2+, turn-off fluorometric for Hg2+ ion, and turn-on fluorogenic sensor for F- ion with the practical application.

A colorimetric and fluorometric probe (E)-2-((8-hydroxy-1,2,3,5,6,7-hexahydropyrido[3,2,1-ij]quinolin-9-yl)methylene)hydrazinecarbothioamide based on thiosemicarbazide and julolidine moieties has been synthesised in pure crystalline form and characterized by 1H NMR, UV-vis, elemental analysis and single crystal XRD. The probe functioned as multitarget ion sensor, detect biologically important metal ions Hg2+ and Mn2+ in dual channel mode. Meanwhile, in mixed solvent media DMF/H2O [8:2], probe displayed selectivity for Hg2+ over other cations by the emission spectrum. Interestingly probe has been explored to recognize F- anion in DMF through ESIPT mechanism. The 1:1 binding stoichiometry of probe with Hg2+ and Mn2+ is confirmed by Job's plot through emission titration and UV-vis titration respectively. Probe is selective and sensitive to Hg2+ and Mn2+ with detection limit as low as 15µM and 0.2µM respectively. The sensing mechanism for selective ions was also scrutinized using 1H NMR experiments and computational studies.