Development and standardization of Indian Alcohol Photo Stimuli (IAPS) for cue-reactivity paradigms in patients with alcohol use disorder.

Background: The availability of appropriate alcohol-related stimuli is a crucial concern for the evaluation and treatment of patients with alcohol dependence syndrome. The study aimed to standardize alcohol-related images with cultural relevance to the Indian setting. Methods: We produced an extensive database of 203 pictures, the Indian Alcohol Photo Stimuli (IAPS), portraying different categories and types of alcoholic beverages, after removing the confounding effects of low-level stimulus parameters (e.g. brightness and blurriness). Thirty patients with alcohol dependence syndrome, currently abstinent, rated each image on visual analog scale (VAS) ranging from 0 (no craving) to 10 (extreme), to determine how typical the stimuli served as craving-relevant stimuli. Results: The mean VAS scores across beverages (ordered from highest to lowest) were whiskey >rum >beer >wine >vodka. Repeated-measures analysis of variance (ANOVA) showed a significant difference in mean VAS scores across beverages (F = 2.93, df = 2.9/86.3, P = 0.039, Greenhouse-Geisser corrected); the effect size for the difference was small (ηp2 = 0.092). A post hoc Bonferroni shows significantly higher VAS scores with whiskey compared with vodka (P = 0.029), whereas the scores were similar across other beverages. A two-way repeated-measures ANOVA for interaction between type of alcoholic beverages and activity was not significant (F = 2.67, df = 2.6/76.6, P = 0.061, Greenhouse-Geisser corrected). Conclusions: We created a standardized alcohol-related image database for studying cue-reactivity paradigms in individuals with alcohol use disorder (AUD). Further research is needed to validate the impact of image features on cue reactivity.

Cognitive function and heart rate variability in open and closed skill sports.

OBJECTIVES: The differences in sporting environments between open and closed skill sports impose unique demands on athletes' cognitive and motor capabilities. Our study aims to investigate and compare cognitive function and Heart Rate Variability (HRV) among individuals involved in different sports, namely basketball, swimming, and a sedentary non-sports group. MATERIALS AND METHODS: The study consisted of three groups, namely basketball players, swimmers, and sedentary individuals, with each group comprising twenty-six participants. HRV was assessed with the help of PowerLab. Cognition was assessed using the Ebbinghaus Memory Procedure Test (EMT), Go/No-Go Task (GNG), Color Stroop task, Trail Making Test (TMT), and Letter Cancellation test (LCT). RESULTS: The results of the Multivariate Analysis of Covariance (MANCOVA) analyses indicated that there was significance between the groups. However, no significant differences were observed between swimmers and basketball players in cognitive functions and HRV measures. Overall, the sport group outperformed the sedentary group. Specifically, basketball players and swimmers completed LCT and TMT faster than the sedentary group (p = 0.044 and p < 0.001 for basketball players, p = 0.002 and p = 0.001 for swimmers). Additionally, basketball players took fewer trials in EMT (p = 0.013) and less time (p = 0.026) compared to the sedentary group. CONCLUSION: The results of the study indicate that sports training, regardless of sport type, positively impacts overall cognitive function. However, no significant differences were observed in cognitive task performance and HRV measures between open and closed skill sport players. These findings suggest that sports can enhance cognitive functions, regardless of the sport played.

Epigenetic signaling and crosstalk in regulation of gene expression and disease progression.

Chromatin modifications - including DNA methylation, modification of histones and recruitment of noncoding RNAs - are essential epigenetic events. Multiple sequential modifications converge into a complex epigenetic landscape. For example, promoter DNA methylation is recognized by MeCP2/methyl CpG binding domain proteins which further recruit SETDB1/SUV39 to attain a higher order chromatin structure by propagation of inactive epigenetic marks like H3K9me3. Many studies with new information on different epigenetic modifications and associated factors are available, but clear maps of interconnected pathways are also emerging. This review deals with the salient epigenetic crosstalk mechanisms that cells utilize for different cellular processes and how deregulation or aberrant gene expression leads to disease progression.

KDM5A noncanonically binds antagonists MLL1/2 to mediate gene regulation and promotes epithelial to mesenchymal transition.

Differential expression of genes involved in certain processes is a collaborative outcome of crosstalk between signalling molecules and epigenetic modifiers. In response to environmental stimulus, interplay between transcription factors and epigenetic modifiers together dictates the regulation of genes. MLLs and KDM5A are functionally antagonistic proteins, as one acts as a writer and the other erases the active chromatin mark, i.e., H3K4me3. KDM5A influences the process of EMT by binding to both epithelial and mesenchymal gene promoters. Through this work, we show that when bound to E-cadherin promoter, KDM5A acts as a classical repressor by demethylating H3K4me3, but on mesenchymal markers, it acts as a transcriptional activator by inhibiting the activity of HDACs and increasing H3K18ac. Further, through our chromatin immunoprecipitation experiments, we observed a co-occupancy of KDM5A with MLLs, we tested whether KDM5A might physically interact with MLLs and WDR5, and here we provide experimental evidence that KDM5A indeed interacts with MLLs and WDR5.

Mechanisms of hedgehog, calcium and retinoic acid signalling pathway inhibitors: Plausible modes of action along the MLL-EZH2-p53 axis in cellular growth control.

Understanding the molecular mechanism(s) of small compounds in cellular growth control are essential for using those against the disease(s). Oral cancers exhibit a very high mortality rate due to higher metastatic potential. Aberrant EGFR, RAR, HH signalling, enhanced [Ca2+] and oxidative stress are some of the important characteristics of oral cancer. So, we target these for our study. Herein, we tested the effect of fendiline hydrochloride (FH) as an LTCC Ca2+-channel inhibitor, erismodegib (a SMO inhibitor of HH-signalling) and all-trans retinoic acid (RA) inducer of RAR signalling that causes cellular differentiation. OCT4 activating compound (OAC1) counters differentiation and induces stemness properties. Cytosine ß-D arabinofuranoside (Cyto-BDA), a DNA replication inhibitor was used to reduce high proliferative capacity. Treatment of FaDu cells with OAC1, Cyto-BDA and FH increase G0/G1 population by 3%, 20% and 7% respectively, and lead to reduction of cyclin D1, CDK4/6 levels. Erismodegib arrests the cells in S-phase with reduced cyclin-E1&A1 levels, whereas RA-treatment causes G2/M phase arrest with reduced cyclin-B1. There was a decrease in the expression of EGFR and mesenchymal markers, Snail/Slug/Vim/Zeb/Twist, and increased E-cadherin expression in all the drug treatments, indicating a reduction in proliferative signal and EMT. Enhanced MLL2 (Mll4) and reduced EZH2 expression associated overexpression of p53 and p21 were traced out. We conclude that these drugs impact expression of epigenetic modifiers by modulating signalling pathways and the epigenetic modifiers then controls the expression of cell cycle control genes, including p53 and p21.

Enhancing our ability to diagnose cardiac valve disease by applying a graphical educational game.

Objectives: This study aimed to compare a newly designed graphical educational game (GEG) with a case-based learning (CBL) exercise and to enhance our ability to apply physiological knowledge of the cardiac cycle to diagnose cardiac valvular diseases among preclinical medical students. Methods: In this interventional study, first-year undergraduate medical students were randomly assigned to a GEG group (n = 42) and a CBL group (n = 37). The GEG group involved shading cardiac cycle graphs and pressure-volume loops while the CBL group worked on two cases of cardiac valve diseases. A multiple-choice question (MCQ) test was then used to assess conceptual understanding of the cardiac cycle. After brief exposure to murmur auscultation on a simulator manikin, the groups were assessed in a simulator manikin test for their ability to diagnose cardiac valve disease. Median MCQ scores and mean scores in the simulator test were then compared using the Mann-Whitney U test. The student's perspectives of the GEG and simulation session were acquired on a 5-point Likert scale questionnaire. Results: The GEG group had significantly higher median MCQ scores (p < 0.001) and mean simulator test scores (p < 0.001) when compared to the CBL group. Moreover, 91% of students agreed that the GEG helped them to clarify concepts, and 88% agreed that the concepts and knowledge gained through the GEG helped them to diagnose valve disease in the manikins. Conclusion: The GEG was positively received by students and was more useful than the CBL in enhancing the application of cardiac physiology concepts and improving diagnostic ability in a simulated clinical setting.

PAX9 reactivation by inhibiting DNA methyltransferase triggers antitumor effect in oral squamous cell carcinoma.

Aberrant DNA hypermethylation is associated with oral carcinogenesis. Procaine, a local anesthetic, is a DNA methyltransferase (DNMT) inhibitor that activates anticancer mechanisms. However, its effect on silenced tumor suppressor gene (TSG) activation and its biological role in oral squamous cell carcinoma (OSCC) remain unknown. Here, we report procaine inhibited DNA methylation by suppressing DNMT activity and increased the expression of PAX9, a differentiation gene in OSCC cells. Interestingly, the reactivation of PAX9 by procaine found to inhibit cell growth and trigger apoptosis in OSCC in vitro and in vivo. Likely, the enhanced PAX9 expression after exposure to procaine controls stemness and differentiation through the autophagy-dependent pathway in OSCC cells. PAX9 inhibition abrogated procaine-induced apoptosis, autophagy, and inhibition of stemness. In OSCC cells, procaine improved anticancer drug sensitivity through PAX9, and its deficiency significantly blunted the anticancer drug sensitivity mediated by procaine. Additionally, NRF2 activation by procaine facilitated the antitumor response of PAX9, and pharmacological inhibition of NRF2 by ML385 reduced death and prevented the decrease in the orosphere-forming potential of OSCC cells. Furthermore, procaine promoted antitumor activity in FaDu xenografts in athymic nude mice, and immunohistochemistry data showed that PAX9 expression was significantly enhanced in the procaine group compared to the vehicle control. In conclusion, PAX9 reactivation in response to DNMT inhibition could trigger a potent antitumor mechanism to provide a new therapeutic strategy for OSCC.

A combined stimuli presentation for assessing facial emotion related N170, N250 and visual mismatch negativity in neuropsychiatric disorders.

Event-related potential (ERP) is a useful approach to assess the neurophysiological correlates of facial emotion processing. Previous studies examined the facial emotion recognition (FER) related ERPs (N170, N250, visual MisMatch Negativity) individually using ERP specific paradigms. This approach can be time-consuming and may not resemble real-life scenarios where an individual must process multiple stimuli simultaneously. The aim of the study was to assess the utility of a combined paradigm when compared to individual paradigms to measure N170, N250 and visual MisMatch Negativity (vMMN) in healthy controls (HC), utilizing emotion stimuli standardized in the Indian population. Further, the combined paradigm was examined in patients with schizophrenia (SCZ) to detect the differences in ERPs compared to HC. Within paradigms, ERPs showed higher amplitudes for emotion compared to neutral stimuli suggesting that the paradigms were able to detect valence associated with emotional stimuli. The combined paradigm was able to elicit decipherable peaks of N170, N250 and vMMN similar to individual paradigms. ERP data quality as assessed by analytic Standardized Measurement Error (aSME) showed a satisfactory aggregate score of above 2 for all the three paradigms. Combined paradigm approaches to record ERPs in neuropsychiatric conditions has the advantage of reducing the time required for task administration, avoiding practice effects, better subject cooperation and participation.

One-month long foundation course for newly joined Indian medical undergraduates: Faculty and students' perspective.

BACKGROUND: A one-month long foundation course has been introduced at the entry-level for first-year MBBS (Bachelor of Medicine and Bachelor of Surgery) students in the medical institutions across India from 2019. Therefore, the present study is aimed at describing the experience of implementing a one-month long foundation course conducted for the Competency-based Undergraduate Medical Curriculum (CBUC) of Indian Medical Graduate as per the guidelines from the National Medical Commission (NMC) (erstwhile Medical Council of India, MCI). We have evaluated the student and faculty perceptions towards the effectiveness of the program. METHODS: The foundation course had six modules Orientation, Skills, Field visit to Community Health Centre, Professional Development including Ethics, Sports and Extracurricular activities, Computer Skills, and Language enhancement program. Regular feedback wascollected from students (N = 250) and teachers (N = 26) involved in the Foundation course using a semi-structured questionnaire. The program's overall feedback was also obtained at the end of the course, using a validated questionnaire. The quantitative findings were expressed in frequency and percentage. The qualitative observations (reflections of students and faculty) were subjected to thematic. RESULTS: The students and faculty appreciated the one-month long foundation course. The course's defined objectives were met as indicated by most students (98.4%) and faculty (75%). The course seemed to be useful for students to embark on a formal MBBS curriculum. It also exposed them to new knowledge and practices, as indicated by the feedback. Thematic analysis of the students' and faculty's reflections was carried out and two themes were identified, i.e., 'strengths' and 'challenges.' The Foundation Course Committee will work out appropriate remedial measures to overcome the challenges in the future sessions for subsequent batches. CONCLUSIONS: The one-month-long foundation course was found to be beneficial for newly joined students to get introduced and adjusted to higher education systems' demands. Also, the challenges faced during the program needs to be addressed with suitable remedial measures while implementing for subsequent batches. This effort will ensure a smooth conduct of the foundation course for the future batches of medical undergraduates and make the program more effective.

Dissecting miRNA facilitated physiology and function in human breast cancer for therapeutic intervention.

MicroRNAs (miRNAs) are key epigenomic regulators of biological processes in animals and plants. These small non coding RNAs form a complex networks that regulate cellular function and development. MiRNAs prevent translation by either inactivation or inducing degradation of mRNA, a major concern in post-transcriptional gene regulation. Aberrant regulation of gene expression by miRNAs is frequently observed in cancer. Overexpression of various 'oncomiRs' and silencing of tumor suppressor miRNAs are associated with various types of human cancers, although overall downregulation of miRNA expression is reported as a hallmark of cancer. Modulations of the total pool of cellular miRNA by alteration in genetic and epigenetic factors associated with the biogenesis of miRNA machinery. It also depends on the availability of cellular miRNAs from its store in the organelles which affect tumor development and cancer progression. Here, we have dissected the roles and pathways of various miRNAs during normal cellular and molecular functions as well as during breast cancer progression. Recent research works and prevailing views implicate that there are two major types of miRNAs; (i) intracellular miRNAs and (ii) extracellular miRNAs. Concept, that the functions of intracellular miRNAs are driven by cellular organelles in mammalian cells. Extracellular miRNAs function in cell-cell communication in extracellular spaces and distance cells through circulation. A detailed understanding of organelle driven miRNA function and the precise role of extracellular miRNAs, pre- and post-therapeutic implications of miRNAs in this scenario would open several avenues for further understanding of miRNA function and can be better exploited for the treatment of breast cancers.

Molecular mechanisms of KDM5A in cellular functions: Facets during development and disease.

Gene expression is influenced at many layers by a fine-tuned crosstalk between multiple extrinsic signalling pathways and intrinsic regulatory molecules that respond to environmental stimuli. Epigenetic modifiers like DNA methyltransferases, histone modifying enzymes and chromatin remodellers are reported to act as triggering factors in many scenarios by exhibiting their control over most of the cellular processes. These epigenetic players can either directly regulate gene expression or interact with some effector molecules that harmonize the expression of downstream genes. One such epigenetic regulator which exhibits multifaceted regulation over gene expression is KDM5A. It is classically a transcriptional repressor acting as H3K4me3 demethylase, but also is reported to act as an activator in many contexts either by loss of activity due to inhibition manifested by other interacting proteins or by downregulating the negative players of a given physiological process thereby escalating the framework. Through this review, we draw attention to the remarkable modes of functioning laid by KDM5A on transcriptional and translational processes, affecting gene expression during differentiation and development and finally summing up on role in disease causation (Fig. 1). We also shed light on different orthologs of KDM5A and their organism specific roles, along with comparison of the sequence similarity to extrapolate some unanswered questions about this protein.

Epigenetic silencing of genes enhanced by collective role of reactive oxygen species and MAPK signaling downstream ERK/Snail axis: Ectopic application of hydrogen peroxide repress CDH1 gene by enhanced DNA methyltransferase activity in human breast cancer.

Loss of E-cadherin and epithelial to mesenchymal transition (EMT) are key steps in cancer progression. Reactive oxygen species (ROS) play significant roles in cellular physiology and homeostasis. Roles of E-cadherin (CDH1), EMT and ROS are intriguingly illustrated in many cancers without focusing their collective concert during cancer progression. We report that hydrogen peroxide (H2O2) treatment modulate CDH1 gene expression by epigenetic modification(s). Sublethal dosage of H2O2 treatment decrease E-cadherin, increase DNMT1, HDAC1, Snail, Slug and enrich H3K9me3 and H3K27me3 in the CDH1 promoter. The effect of H2O2 was attenuated by ROS scavengers; NAC, lupeol and beta-sitosterol. DNMT inhibitor, AZA prevented the H2O2 induced promoter-CpG-island methylation of CDH1. Treatment of cells with U0126 (inhibitor of ERK) reduced the expression of DNMT1, Snail and Slug, increased CDH1. This implicates that CDH1 is synergistically repressed by histone methylation, DNA methylation and histone deacetylation mediated chromatin remodelling and activation of Snail and Slug through ERK pathway. Increased ROS leads to activation of epigenetic machineries and EMT activators Snail/Slug which in their course of action inactivates CDH1 gene and lack of E-cadherin protein promotes EMT in breast cancer cells. ROS and ERK signaling facilitate epigenetic silencing and support the fact that subtle increase of ROS above basal level act as key cell signaling molecules. Free radical scavengers, lupeol and beta-sitosterol may be tested for therapeutic intervention of breast cancer. This work broadens the amplitude of epigenome and open avenues for investigations on conjoint effects of canonical and intrinsic metabolite signaling and epigenetic modulations in cancer.