Investigating the Behavioural and Molecular Functions of <i>Cry1</i> and <i>Cry2</i> Using Mouse Mutants

Endogenous circadian clocks generate rhythms of physiology and behaviour that are synchronised to the environment, principally through the light-dark cycle. In mammals, the circadian clock is dependent on interlocked feedback loops that involve several clock elements such as cryptochromes (Cry1 and Cry2). Post-translational modifications control intracellular trafficking, functionality and degradation of CRY proteins which are keys to the functioning of the clock. CRY protein levels are dependent upon their timely degradation by F-box proteins. This has recently been shown in the afterhours (Afh) mutant carrying a mutation in the F-box gene, Fbxl3. Afh has been shown to lengthen circadian period by stabilising levels of CRY proteins across the circadian cycle. To understand the specific roles of each of the two CRY proteins in circadian regulation, we generated compound mouse mutants to investigate the behavioural and molecular consequences of stabilising either CRY1 or CRY2 protein levels in mice lacking the alternative form of Cry. The circadian wheel-running activity assessed in light:dark and constant environmental conditions for both Cry1-/-;Fbx13Afh/Afh and Cry2-/-;Fbx13Afh/Afh (stabilising CRY2 and CRY1 protein levels respectively); clearly show a gradual increase in period length in constant darkness as the dosage of Fbxl3Afh is increased. This would suggest that stabilisation of either CRY protein can lengthen the clock, presumably as a result of a prolonged phase of transcriptional repression by either protein. This effect seen in the compound mutants was confirmed at the gene and protein levels and it was concluded that Cry1 and Cry2 can both act as transcriptional repressors, but that Cry1 plays the predominant inhibitory role in the cerebellum and peripheral organs. Subsequently it has been shown that FBXL21, the closest homologue of FBXL3, also binds to CRY1 and impairs its repressive action towards the transcriptional activators, CLOCK-BMAL1 presumably by degrading CRY1. Due to differences in their expression Fbxl3 and Fbxl21 may have overlapping roles. In-vitro and in vivo analysis in mutants generated in Fbxl21; revealed CRY2 as a preferable target of FBXL21 and that this may contribute to the lower repressive function of Cry2. Further investigation into the genetic interactions between the two F-box genes showed that Fbxl3 is epistatic to Fbxl21. Finally, Fbxl21 has been shown to be associated with schizophrenia in humans. In our hands mutant mouse Fbxl21 showed no such associations, instead indicating an association with anxiety and/or defects in sensorimotor gating

Genetically engineered ascorbic acid-deficient live mutants of Leishmania donovani induce long lasting protective immunity against visceral leishmaniasis

Visceral leishmaniasis caused by Leishmania donovani is the most severe systemic form of the disease. There are still no vaccines available for humans and there are limitations associated with the current therapeutic regimens for leishmaniasis. Recently, we reported functional importance of Arabino-1, 4-lactone oxidase (ALO) enzyme from L. donovani involved in ascorbate biosynthesis pathway. In this study, we have shown that ΔALO parasites do not affect the ability of null mutants to invade visceral organs but severely impair parasite persistence beyond 16 week in BALB/c mice and hence are safe as an immunogen. Both short term (5 week) and long term (20 week) immunization with ΔALO parasites conferred sustained protection against virulent challenge in BALB/c mice, activated splenocytes and resulted in induction of pro-inflammatory cytokine response. Protection in immunized mice after challenge correlated with the stimulation of IFN-γ producing CD4+ and CD8+ T cells. Antigen-mediated cell immunity correlated with robust nitrite and superoxide generation, macrophage-derived oxidants critical in controlling Leishmania infection. Our data shows that live attenuated ΔALO parasites are safe, induce protective immunity and can provide sustained protection against Leishmania donovani. We further conclude that the parasites attenuated in their anti-oxidative defence mechanism can be exploited as vaccine candidates

Essential Skills in Postgraduate Medical Curriculum of Community Medicine

Introduction: Community-based education has been considered a suitable approach for health promotion and for requisite skill development regarding primary health care. In the current perspective, public health training and research, being two important aspects require immediate attention.Objective: To assess the skills of Postgraduate Students in the Department of Community Medicine in four Medical Colleges of Delhi.Materials and Methods: It was a cross-sectional study conducted among 70 Postgraduate Medical Students of 4 Medical Colleges in Delhi. The data were collected through a self administered, pre- tested questionnaire containing items assessing socio-demographic profile and skills essential for Postgraduate Students of Community Medicine.Results: There were 58.6% male and 29% female students. A large proportion of participants were having age range between 25-29 years. Ability 'to resolve conflict among the nurse at Primary Health Centre (PHC)', 'generate community participation', 'making thick and thin smear in case of fever', 'making a chart showing month-wise distribution of CuT', and 'calculating Chi-square of data', were found to significantly higher in 2nd and 3rd year PG students than first year PG students (p&lt;0.01). Only 27.1% of students felt that they could test water sample for microbiological aspects while only 47.1% said that they could examine an industrial worker for pre-placement examination.Conclusions: PG students assessed themselves to possess necessary skills on communication, counselling and health education. However, many students lacked skills pertaining to occupational health and epidemiology.Keywords: Competency in community medicine, public health, epidemiological skills, communication skills

Evaluation of Preclinical Task Based Learning program in Medical Education [version 1; peer review: 1 approved, 2 approved with reservations]

The conventional curriculum in preclinical medical education has a need for early clinical exposure programs that help in correlation of basic science data with clinical skills. This is helpful to develop clinical reasoning skills, problem-solving abilities, team work, communication skills and overall attitudes and behaviour relevant for a healthcare provider. Preclinical task based learning (TskBL) is an active learning strategy in which the focus for the first year medical student is a real task done by a doctor. In this strategy the student-doctors undergo a standardized patient encounter and discuss the learning issues related to the task in the first year of medical school. The current study is focussed on the student perception of the effectiveness of task based learning module.The TskBL was conducted among first year medical students for nine topics that are commonly encountered in the clinics. After TskBL was planned and implemented the evaluation of the modules was done using focus group discussions. The students highlighted the importance of standardized patients in the TskBL strategy in providing early clinical exposure in preclinical medical education. They reported its usefulness gaining essential knowledge, skills and attitudes for medical learning. They reported positive outcomes of module design and processes and activities in TskBL. Based on the negative aspects of the modules, future improvement was suggested in improving the usefulness of standardized patient encounter. This study showed the novice learners’ outlook of the potency of TskBL for several other topics of clinical relevance to provide early clinical exposure in medical schools

Role of comprehensive diabetes care in known diabetes patients from western Mumbai region: an observational study

Background: The occurrence of Diabetes Mellitus (DM) has been creating a havoc since past few decades on a global platform. As per available literature, prevalence of DM in Mumbai is around 10%. Comprehensive Diabetes Care (CDC) is a form of Ayurvedic therapy which combines panchkarma and diet management. The present study was planned to evaluate the effectiveness of CDC in patients of DM by analysing changes in body mass index (BMI), body weight, OGTT, HbA1c, etc.Methods: The present study was of retrospective design, conducted at Madhavbaug clinics in western Mumbai. The duration of study was of one year, conducted from October 2018 to September 2019. It included patients diagnosed with type 2 DM i.e. HbA1c>6.5%, who were given CDC therapy.Results: In the present study, out of 183 type 2 diabetic patients, 99 were males (52%), while 84 were females (48%), thus male: female ratio was 1.17:1. On analysing the results of HbA1c in patients who had completed 12 weeks of CDC therapy, it was found that controlled DM status was seen in 109 patients (59%), while uncontrolled DM status was noted in 33 patients (19%) as compared to 102 patients (58%) at baseline.Conclusions: From the findings of the present study, it is clear that CDC is effective in the form of increasing number of euglycemic patients at the end of study period, as well as reduction in all glycaemic and anthropometric parameters, and reducing dependency on conventional medicines

aDolescents gEnder surVey, rEsponsible coupLes evaluatiOn, and capacity building Project in India (DEVELOP): a study protocol

Gender-based violence against women is a serious concern in India. This affects the health and wellbeing of victims and their dependents, such as children. Published evidence has documented a variety of reasons for such violence in Indian societies, paving a pathway to design, implement, and evaluate intervention models to address this issue. DEVELOP is a research study designed by UK and Indian research teams to plan future projects to address gender-based discrimination and violence against women and girls in India. This study protocol provides detailed information on the objectives, research methods, data collection, storage, analysis, and dissemination plans of the DEVELOP. The first component is a survey of adolescent boys and girls from rural areas of Maharashtra state of India to understand their gender equality related knowledge and beliefs. The insight gathered will be used to design interventions targeted at adolescent populations through future research and development programmes. Secondly, an evaluation of the ‘Responsible Couples’ project will be conducted to assess its success and challenges, and to design suitable programme activities and models. The ‘Responsible Couples’ project is implemented in 40 villages of Maharashtra state to improve relationships in married couples, prevent violence against women, intervene during violence, and to provide support services for women and their family members. Research findings will be disseminated though public engagement events in India, international conferences, and peer reviewed publications. This will impact on the policy and work models of Indian partners to plan future project investments. Research findings will be also useful for local government authorities and other non-government agencies working on gender equality themes