Folic acid depletion along with inhibition of the PERK arm of endoplasmic reticulum stress pathway promotes a less aggressive phenotype of hepatocellular carcinoma cells.

Folate is a vital vitamin involved in one-carbon metabolism and any changes in folate status may lead to epigenetic alterations. It is already known that stages and liver cancer progression are negatively correlated with folate levels. Nevertheless, mechanisms involved in folate deficiency in HCC (Hepatocellular carcinoma) are still not completely understood. So, this study tests the hypothesis that due to the increased demand for ER (endoplasmic reticulum) proteins, folate deficiency might lead to the induction of UPR (unfolded protein response), which is further correlated with HCC outcomes. HCC cells were cultured in both folate normal (FN) and folate deficient (FD) conditions and the expression of genes of ER stress pathway was investigated. The results demonstrated activation of UPR via induction of PERK, ATF4, and LAMP3. Besides this, FD reduced the migratory capacity and the invasiveness of HCC cells along with the reduction in mesenchymal markers like vimentin but increased apoptosis. Treatment with GSK2606414 (PERK inhibitor) decreased the FD induced expression of PERK, ATF4, and LAMP3 in FD cells. Also, GSK2606414 was found to increase apoptotic cell death and to further reduce the cancer hallmarks selectively in FD cells but not in FN cells. Altogether, our data suggest that targeting the ER stress pathway along with folate deficiency may provide a more promising elimination of the metastatic potential of HCC cells contributing to more effective therapeutic agents.

Reassessing the availability of crop residue as a bioenergy resource in India: A field-survey based study.

Second-generation bioenergy, a carbon neutral or negative renewable resource, is crucial to achieving India's net-zero emission targets. Crop residues are being targeted as a bioenergy resource as they are otherwise burned on-field, leading to significant pollutant emissions. But estimating their bioenergy potential is problematic because of broad assumptions about their surplus fractions. Here, we use comprehensive surveys and multivariate regression models to estimate the bioenergy potential of surplus crop residues in India. These are with high sub-national and crop disaggregation that can facilitate the development of efficient supply chain mechanisms for its widespread usage. The estimated potential for 2019 of 1313 PJ can increase the present bioenergy installed capacity by 82% but is likely insufficient alone to meet India's bioenergy targets. The shortage of crop residue for bioenergy, combined with the sustainability concerns raised by previous studies, imply a need to reassess the strategy for the use of this resource.

Tumor suppressor genes are differentially regulated with dietary folate modulations in a rat model of hepatocellular carcinoma.

The current study evaluated the outcome of dietary folate modulations on the expression of tumor suppressor genes (TSGs) during developmental stages of hepatocellular carcinoma (HCC) in a Wistar rat model. In addition to dietary folate modulations, male rats were administered diethylnitrosamine (DEN) intraperitoneally once a week upto 18 weeks to induce HCC. Serum folate levels were found to be decreased and increased in folate deficiency (FD) and folate-oversupplemented (FO) groups respectively when compared to folate normal (FN) rats. Apoptosis was observed in FD in fibrosis and HCC stages. mRNA expression analysis by RT-PCR of TSGs (DPT, p16, RUNX3, RASSF1A and SOCS1) and protein expression by western blot (RASSF1A, RUNX3 and p16) depicted differential expression in FD and FO in various stages of HCC development. Bisulfite sequencing for p16 and RASSF1A promoter was performed. The promoter region of p16 gene was hypermethylated at 7th and that of RASSF1A was hypomethylated at 10th CpG in cirrhotic category in FD rats. Hyper and hypomethylation at 10th and 24th CpG respectively in RASSF1A promoter was observed in HCC category in both FD and FO groups. All TSGs showed differential expression at transcript and protein level. Increased expression of DPT, RASSF1A, SOCS1 and decreased expression of RUNX3 could be playing role in HCC development in FD rats. Reduced expression of RUNX3, RASSF1A and SOCS1 in HCC category was demonstrated in FO rats. Thus, the studied TSGs are differentially expressed with dietary folate modulations during the development of HCC in DEN-treated rat model and the promoter methylation might be a contributing mechanism under these conditions.

Association of interleukin-18 genotypes (-607C > A) and (-137 G > C) with the hepatitis B virus disease progression to hepatocellular carcinoma.

Chronic infection with HBV has been reported to be associated with the development of HCC. The inflammation mounted by cytokine-mediated immune system plays an important role in the pathogenesis of HBV-associated HCC. IL-18 is a pro-inflammatory cytokine whose role in the development of HBV-associated chronic to malignant disease state has not been much studied. The present study was conceived to determine the role of genetic polymorphisms in IL-18, serum levels of IL-18, and expression level of its signal transducers in the HBV disease progression. A total of 403 subjects were enrolled for this study including 102 healthy subjects and 301 patients with HBV infection in different diseased categories. Polymorphism was determined using PCR-RFLP. Genotypic distributions between the groups were compared using odd's ratio and 95% CI were calculated to express the relative risk. Circulating IL-18 levels were determined by ELISA. Expression levels of pSTAT-1 and pNFƙB was determined by western blotting. In case of IL-18(- 607C > A), the heterozygous genotype (CA) was found to be a protective factor while in case of IL-18(- 137G > C) the heterozygous genotype (GC) acted as a risk factor for disease progression from HBV to HCC. Moreover, serum IL-18 levels were significantly increased during HBV disease progression to HCC as compared to controls. Also the levels of activated signal transducers (pSTAT-1 and pNF-κB) of IL-18 in stimulated PBMCs were significantly increased during HBV to HCC disease progression. These findings suggest that IL-18 has the potential to act as a biomarker of HBV-related disease progression to HCC.

Comparison of two formats of journal club for postgraduate students at two centers in developing critical appraisal skills.

Journal club (JC) is an integral part of postgraduate medical education. Although several innovations have been attempted to improve its effectiveness, the influence of the number of sessions remains unexplored. The current study investigated the effect of the number of sessions (one vs. four) of conduction of journal club on critical appraisal skills of postgraduate students at two institutions. A quasi-experimental study was conducted comparing the effectiveness of JC in a one- versus four-session JC format using two objective tests: Critical Appraisal Skills Test (CAST) and Population, Intervention, Comparison, Outcome (PICO) score. The degree of changes (pre-JC vs. post-JC session) after each format exposure, association of prescores to postscores, and correlation between the CAST and PICO scores were discerned. CAST score was higher in the four-session format (P = 0.03), whereas PICO score increased in the single-session format of JC (P = 0.03). Also, the change in CAST score was higher in the four-session versus single-session JC format (P = 0.04). We conclude that a four-session format of JC is more effective in augmenting critical appraisal skills of postgraduate students, whereas a single session is sufficient for improving factual knowledge.

IUPS Physiology Education Workshop series in India: organizational mechanics, outcomes, and lessons.

Active learning promotes the capacity of problem solving and decision making among learners. Teachers who apply instructional processes toward active participation of learners help their students develop higher order thinking skills. Due to the recent paradigm shift toward adopting competency-based curricula in the education of healthcare professionals in India, there is an emergent need for physiology instructors to be trained in active-learning methodologies and to acquire abilities to promote these curriculum changes. To address these issues, a series of International Union of Physiological Sciences (IUPS) workshops on physiology education techniques in four apex centers in India was organized in November 2018 and November 2019. The "hands-on" workshops presented the methodologies of case-based learning, problem-based learning, and flipped classroom; the participants were teachers of basic sciences and human and veterinary medicine. The workshop series facilitated capacity building and creation of a national network of physiology instructors interested in promoting active-learning techniques. The workshops were followed by a brainstorming meeting held to assess the outcomes. The aim of this report is to provide a model for implementing a coordinated series of workshops to support national curriculum change and to identify the organizational elements essential for conducting an effective Physiology Education workshop. The essential elements include a highly motivated core organizing team, constant dialogue between core organizing and local organizing committees, a sufficient time frame for planning and execution of the event, and opportunities to engage students at host institutions in workshop activities.

Imbalance between matrix metalloproteinases and their tissue inhibitors in preeclampsia and gestational trophoblastic diseases.

The invasion cascade exhibited by placental trophoblasts and cancerous cells bears many similarities, and it is attributed to extracellular matrix degradation mediated by matrix metalloproteinases (MMPs). Although proper and controlled invasion by trophoblasts into the maternal uterus is an essential requirement for maintenance of normal pregnancy, any abnormality in this phenomenon results in the development of invasion-related disorders such as gestational trophoblastic diseases (GTDs) and preeclampsia. We studied the epigenetic basis of differential expression of two placental MMPs (MMP2 and MMP9) and tissue inhibitors of metalloproteinases (TIMP2 and TIMP1) during normal gestation and invasion-related disorders, i.e., preeclampsia and GTDs. Our study suggests the association of H3K9/27me3 with differential expression of these MMPs and their inhibitors, which regulate the placental invasion during normal pregnancy, whereas no role of CpG methylation was observed in the differential expression of MMPs/TIMPs. Further, development of GTDs was associated with abnormally higher expression of these MMPs and lower levels of their inhibitors, whereas the reverse trends were observed for MMPs and their TIMPs in case of preeclampsia, in association with abnormal changes in H3K9/27me3. These results suggest the involvement of higher levels of MMPs in an aggressive invasive behavior depicted by GTDs, whereas lower levels of these MMPs in shallow and poor invasive phenotype associated with preeclampsia. Thus, our study shows the significance of a proper balance regulated by histone trimethylation between differential expression of MMPs and their TIMPs for maintaining normal pregnancy and its deregulation as a contributing factor for pathogenesis of invasive disorders during pregnancy.

Folic acid depletion as well as oversupplementation helps in the progression of hepatocarcinogenesis in HepG2 cells.

Folate ingestion below and above the physiologic dose has been shown to play a tumorigenic role in certain cancers. Also, excessive folate supplementation after establishment of pre-established lesions led to an advancement in the growth of a few tumors. However, such information has not yet been achieved in the case of HCC. In our study, HepG2 cells were administered with three different concentrations of folic acid i.e. folic acid normal (FN) (2.27 µM), folic acid deficient (FD) (no folic acid), folic acid oversupplementation (FO) (100 µM) for 10 days. Intracellular folate levels were assayed by Elecsys Folate III kit based method. The migratory and invasive abilities were estimated by transwell migration and matrigel invasion methods respectively. FACS was done to evaluate cell viability and apoptosis. Agarose-coated plates were used to access cancer stem cells (CSCs) number. Quantitative RT-PCR and western blotting approaches were used for gene and protein expression of certain tumor suppressor genes (TSGs), respectively. FD cells depicted increased migration, invasion, apoptosis, necrosis and decreased cell viability, CSCs. On the other hand, FO cells showed increased migration, invasion, cell viability and number of CSCs and decreased apoptosis and necrosis. TSGs revealed diminished expression with both FA modulations with respect to FN cells. Thus, FA deficiency as well as abundance enhanced the HCC progression by adapting different mechanisms.

Polymeric nanosponges as an alternative carrier for improved retention of econazole nitrate onto the skin through topical hydrogel formulation.

The present study was carried out to exploit the feasibility of using polymeric nanosponges as an alternative carrier for targeting econazole nitrate (EN) to the skin through topical hydrogel formulation. Nanosponges prepared by emulsion solvent diffusion method were evaluated for various physicochemical parameters and in vitro drug release. The nanosponges of EN were discrete free flowing nanosized particles with perforated orange peel like morphology as visualized by SEM. The nanosponge formulated using PVA:EC (3:2) displayed highest in vitro release after 12?h in phosphate buffer (pH 6.8) that fitted matrix model. Selected nanosponge was formulated as Carbopol 934 NF hydrogel using varying concentrations of permeation enhancers propylene glycol and N-methyl-2-pyrrolidone. The EN nanosponge-loaded hydrogels (F0?F7) were evaluated for pharmacotechnical properties and irritation studies on rat skin. On the basis of various evaluation parameters F7 with an equilibrium swelling of 0.944?g/g after 4?h, low firmness and high adhesiveness, a flux rate of 1540.2 (?g/cm(2).h) and that exhibited a controlled release of EN for 12?h was selected as best hydrogel. The DRS and DSC spectra of F7 confirmed stability of drug in the delivery system and absence of drug polymer interaction in nanosponges.

Blunt Abdominal Trauma With Hollow Viscus and Mesenteric Injury: A Prospective Study of 50 Cases.

Introduction Hollow viscus injury following blunt abdominal trauma is an infrequent diagnosis. Blunt hollow viscus and mesenteric injury (HVMI) is not only an uncommon finding but its timely diagnosis is also difficult. Due to its less frequency, this injury has not been studied in detail prospectively. Aims and objectives The aim of this study is to determine the causes, pattern, management, and outcome of HVMI following blunt abdominal trauma. Methodology This study was conducted from January 2015 to June 2016 in a high-volume tertiary care trauma center and teaching hospital in North India. All patients with blunt HVMI admitted during this period were included in this study. Data were collected regarding medical history, physical findings, demographics, injury dates and times, laboratory results, diagnostic tests, delay in surgical intervention, type of surgical procedure performed, site of injury, complications, and mortality. Results Out of a total of 6,570 trauma admissions, 465 blunt abdominal injuries were identified, and HVMI was found only in 50 patients. The small bowel was the most common injury, with the jejunum being the most commonly involved segment. All patients were managed surgically. The mean time to operative intervention after hospital admission was 4.5 hours (IQR: 2-8 hours). Primary repair was performed in 54% of patients. Mortality rate was high in patients with HVMI (22% patients). Septic shock was the most common cause of death. Conclusion Hollow viscus injury in blunt abdominal trauma is not so common finding. Early diagnosis and treatment is an important but difficult task. Prognosis depends on age, associated injuries, co-morbid conditions, and delay in operative intervention.

Dietary modulations of folic acid affect the development of diethylnitrosamine induced hepatocellular carcinoma in a rat model.

The present study evaluated the role of dietary folate modulations in the development of hepatocellular carcinoma (HCC) in a rat model. Male Wistar rats were given diethylnitrosamine (DEN) carcinogen for a period of 18 weeks in addition to different folate modulations. Biochemical parameters were assayed and liver tissues were examined using various histopathological stains viz. Hematoxylin and eosin (H&E), Masson's trichrome, Immunohistochemistry (IHC) staining for arginase-1 and α-smooth muscle actin (SMA). Serum folate and hepatic folate stores were decreased and increased in folate deficiency (FD) and folate oversupplemented (FO) group respectively. Analysis of serum liver function tests revealed deranged liver functioning in all the groups. H&E staining of rat liver demonstrated vague nodularity from 2nd to 8th week, fibrosis from 10th to 15th week, cirrhosis and HCC from 16th to 18th week. Combining the observations of H&E with IHC for arginase-1, 14 (50%), 11 (39.3%) and 17 (58.6%) rats showed HCC positivity in FN (folate normal), FD and FO diets respectively. IHC for α-SMA depicted increased staining with progression of the disease from fibrosis to cirrhosis in all the dietary groups. Collectively, findings of all the histopathological stains, revealed increase in the number of cirrhotic cases and decrease in the number of HCC cases in FD group, indicating delayed progression of HCC with FD. Moreover, FO led to more number of HCC and reduction in the number of cirrhotic cases, signifying early progression of HCC.

Evaluating exercise challenge to validate cardiac autonomic dysfunction in lean PCOS phenotype.

Background Polycystic ovary syndrome (PCOS) is frequently associated with infertility, obesity, hypertension, and diabetes mellitus. Cardiac autonomic dysfunction is well documented in long-term patients of PCOS, with impairment being proportionate to disease duration. However, there is paucity of literature on cardiac autonomic tone in the "lean" phenotype of PCOS. We used exercise challenge to probe for autonomic dysfunction, as assessed by heart rate variability (HRV), an index of cardiac autonomic tone. Methods Our study population consisted of 27 newly diagnosed PCOS patients and 25 healthy females matched by age and body mass index. Short-term HRV was assessed using time and frequency domain indices. Moderate, isotonic exercise was used as an interventional tool. Indices of both groups were compared in three bins - at baseline, immediate, and late postexercise stages. Results The groups had comparable HRV indices at baseline. However, low-frequency (LF) power was significantly reduced in PCOS patients during immediate and late postexercise phases when compared with controls (p = 0.03 and 0.03, respectively). Time domain indices also exhibited a fall postexercise, although not statistically significant. Conclusions Although "lean" phenotype PCOS patients had comparable HRV parameters as controls at baseline, the administration of exercise challenge led to reduced sympathetic drive, evident by reduced LF power in patient group. This may be due to latent autonomic dysfunction in "lean" PCOS, which is unmasked on exposure to exercise challenge. We propose that the evaluation of HRV response to exercise may serve as a sensitive screening tool to detect early cardiovascular dysfunction in newly diagnosed lean PCOS patients.

A new variant of Croton yellow vein mosaic virus naturally infecting wild sunflower in India.

During 2015-2016, wild sunflower (Verbesina encelioides) was observed with bright yellow vein mosaic symptims in Noida, Uttar Pradesh, India. The initial analysis by PCR with a pair of coat protein based primers revealed the association of a begomovirus. Further, the virus was identified by rolling circle amplification and cloning of the complete genome. The DNA-A (2671 nucleotides, MH359168) with the genome organisation typical of the Old World begomovirus shared 98% sequence identity with that of Croton yellow vein mosaic virus (CYVMV) reported from India. The betasatellite (MH359169) associated with the disease shared 92% sequence identity with papaya leaf curl betasatellite (KY825245). The results showed that the wild sunflower is a new alternate host of CYVMV. The study also revealed a natural association of heterologous betasatellite with CYVMV in wild sunflower exhibiting yellow vein mosaic symptoms.

Sleep Pattern of Adolescents in a School in Delhi, India: Impact on their Mood and Academic Performance.

OBJECTIVES: To examine the sleep pattern and observe differences in sleep routines, phase preferences, mood, attendance, and academic performance among different adolescent age students. Secondly, to observe the age at which sleep phase transition and changes in sleep requirement become evident. METHODS: A cross-sectional study was conducted among 501 students (aged 11-15 y) of a school in Delhi, India. Students were evaluated for their sleep patterns, sleep duration, habits of napping, quality of sleep, sleepiness, depression, phase preferences by self-reported school sleep habits survey questionnaire along with school performance and attendance. RESULTS: Significant differences were found in sleep pattern of students aged 11-12 y and 13-15 y. Bedtime shifted to a later time with increasing age but early morning schools kept the wake time same, leading to a decline in total sleep duration of older adolescents. Older adolescents had higher depression but poor attendance and academic performance. Prevalence of sleep deprivation increased with age, from 83.7% to 87.1% in 11-12 y to 90.5% to 92.5% in 13-15 y. CONCLUSIONS: The study clearly identifies 12-13 y as age of transition of sleep pattern among adolescents. Though significant differences were found in the academic performance, mood and attendance among preteens and teens but no direct association was seen between academic performances and sleep pattern. A complex multifactorial association between sleep patterns, attendance, mood and academic performance which may change over days, months, or years should be explored further in a longitudinal follow up study.

A Pilot Study for Evaluation of Digital Systems as an Adjunct to Sphygmomanometry for Undergraduate Teaching.

OBJECTIVES:  Blood pressure estimation is a key skill for medical practitioners. It is routinely taught to undergraduate medical students using an aneroid sphygmomanometer. However, the conceptual understanding in the practical remains limited. We conducted the following study to evaluate the efficacy of digital data acquisition systems as an adjunct to the sphygmomanometer to teach blood pressure. METHODS:  Fifty-seven first-year medical students participated in the study. An MCQ test of 15 questions, consisting of 10 conceptual and five factual questions, was administered twice - pre- and post-demonstration of blood pressure measurement using a digital data acquisition system. In addition, qualitative feedback was also obtained. RESULTS:  Median scores were 7 (6 - 8) and 3 (1.5 - 4) in pre-test sessions for conceptual and factual questions, respectively. Post-test scores showed a significant improvement in both categories (10 (9 - 10) and 4 (4 - 4.5), respectively, Mann-Whitney U test, p < 0.0001). Student feedback also indicated that the digital system enhanced learning and student participation. CONCLUSIONS:  Student feedback regarding the demonstrations was uniformly positive, which was also reflected in significantly improved post-test scores. We conclude that parallel demonstration on digital systems and the sphygmomanometer will enhance student engagement and understanding of blood pressure measurement.

Pilot Study to Assess the Quality of Life, Sleepiness and Mood Disorders among First Year Undergraduate Students of Medical, Engineering and Arts.

INTRODUCTION: College students, who are in a phase of transition from adolescence to adulthood, face numerous challenges. Due to stress overload, students easily fall prey to mood disturbances like stress, anxiety and depression compromising their quality of life. AIM: Present study was undertaken to see the effect of choice of professional course on quality of life and mood of the first year students of medical, engineering and arts students. MATERIALS AND METHODS: A cross-sectional study was conducted amongst 150 students, 50 participants from each stream. Enrolled subjects were administered questionnaires pertaining to their quality of life (WHOQOL-BREF), mood disorders (DASS-42) and sleepiness (ESS). RESULTS: Medical students showed significantly higher levels of stress (p-value=0.0001), depression (p-value=0.002) and anxiety (p-value=0.002), 30% of medicos labelled their quality of life as very good compared to 48% and 50% of engineering and arts students. A 38% of medical student's reported daytime sleepiness compared to 12% engineering and 6% arts students. CONCLUSION: Present study shows that medical students are maximally vulnerable to mood disorders and have a poor quality life. As the student community forms the backbone of any nation, we as, educators should try to strengthen each individual by promoting not only physical and mental development but also aiming for overall holistic development.

Neuroendoscopy for Intracranial Arachnoid Cysts in Infants: Therapeutic Considerations.

Background The use of the endoscope for various cranial procedures is gradually expanding. Intracranial cystic lesions in the brain are one of the most attractive targets for this minimally invasive procedure, thus avoiding conventional craniotomy. These cystic lesions in the brain, namely arachnoid cysts, are congenital. Surgical treatment depends on clinical presentation, location, and age. Patients A total of 13 patients < 1 year of age with intracranial cysts were operated on between 2005 and 2013. Six presented with hydrocephalus, four presented with seizure, one with abnormal head movement, and two had large asymptomatic cysts. Four children had infratentorial arachnoid cysts; of these, three required a transaqueductal procedure. All the patients underwent endoscopic cystoventriculostomy and/or cystocisternostomy and third ventriculostomy in selected cases with a biopsy from the cyst wall. Results Clinically and radiologically all children showed significant improvement with an average follow-up ranging from 8 months to 6 years. There were no intraoperative complications. Three children developed subdural hygroma that subsided with conservative treatment, and one child with pseudomeningocele required a cystoperitoneal shunt at a later date. Conclusion A symptomatic intracranial arachnoid cyst or a large asymptomatic cyst are indications for neurosurgical intervention, and endoscopy is a good treatment option with the advantage of minimal invasiveness and fewer complications. Endoscopic surgery has to be tailored according to the location and presentation.

Effect of Isometric Exercise on QTc Interval.

INTRODUCTION: The QTc interval is affected by heart rate, autonomic nervous system and diseases like diabetes. However, the affect of exercise which alters autonomic nervous system activity, on QTc is not clear. On the other hand, the incidence of sudden cardiac death increases many fold post exercise. These events may be better explained by studying the effect of exercise on QTc. AIM: This study was designed with an aim to record the QTc interval changes in response to isometric exercise in a group of normal individuals with or without parental history of diabetes mellitus. Also the QTc duration was correlated with the LF-HF ratio. MATERIALS AND METHODS: Twenty nine, healthy medical students were subjected to isometric hand grip test for 5min. ECG was recorded pre-exercise and at various time intervals post-exercise. STATISTICAL ANALYSIS: All data are expressed in mean ± SD. Intra group comparison was done using paired t-test and unpaired t-test was used for comparison among group I and group II subjects, and among males and females. RESULT: The difference in the pre and post exercise QTc values both within and between groups was statistically significant with group I subjects recording lower values. The post exercise LF: HF values were significantly increased when compared to pre exercise values in both the groups. There was no correlation between LF: HF and QTc. CONCLUSION: A longer than normal QTc interval predisposes to arrhythmia. Exercise brings about detectable changes in the QTc interval after a period of isometric exercise in normal individuals which in high risk individuals may predispose to sudden cardiac death. In addition women may be more susceptible to post-exercise arrhythmia owing to a longer QTc even at rest.