Investigating the role of an immediate early gene FOS as a potential regulator of autophagic response to hypoglycemia in embryonic hypothalamic neurons.

Hypoglycemia-associated autonomic failure (HAAF) is a well-established complication of diabetes. Although HAAF has serious outcomes such as recurrent morbidity, coma, and death, the mechanisms of HAAF and its pathological components are largely unknown. Our previous studies have revealed that hypoglycemia is associated with the upregulation of an immediate early gene - FOS. In addition, it is documented that glucose deprivation activates neuronal autophagic activities. Therefore, the present study aimed to identify the role of FOS and one of the core components of the autophagy pathway, Beclin-1 (encoded by the BECN1 gene), in the regulation of autophagic mechanisms in embryonic hypothalamic neurons in response to hypoglycemic conditions. Embryonic Mouse Hypothalamic Cell Line N39 (mHypoE-N39 or N39) was cultured in reduced concentrations of glucose (2000, 900, 500, and 200 mg/L). Gene and protein expression, as well as immunofluorescence studies on autophagy were conducted under different reduced glucose concentrations in N39 hypothalamic neurons with and without FOS and BECN1 gene knockdowns (KD). The outcomes of the present study have demonstrated a significant increase in autophagosome formation and subsequent lysosomal degradation in the hypothalamic neurons in response to reduced glucose concentrations. This hypoglycemic response appears to be lowered to a similar extent in the FOS KD and BECN1 KD cells, albeit insignificantly from the negative control, is indicative of the involvement of FOS in the autophagic response of hypothalamic neurons to hypoglycemia. Moreover, the KD cells exhibited a change in morphology and reduced cell viability compared with the control cells. Our findings suggest that reduced FOS expression could potentially be associated with impaired autophagic activities that are dependent on BECN1, which could lead to decreased or blunted hypothalamic activation in response to hypoglycemia, and this, in turn, may contribute to the development of HAAF.

Cryopreservation and validation of differentiated PDGFRα-positive cells for long term usage in experimentation.

OBJECTIVE: Differentiation of immortalized Mesenchymal Stromal Cells (iMSCs) into PDGFRα-positive cells under controlled growth conditions has several vital implications in functional studies concerned with the pathogenesis of Diabetic Gastroparesis (DGP). A study published previously by our research group demonstrated the importance of these cells as a novel, in-vitro model for investigating the functional role of neuronal nitric oxide synthase. The currently available methods require fresh differentiation of PDGFRα-positive cells for each round of experimentation. This leads to longer delays, higher usage of reagents, and inconsistency in reproducibility of experiments frequently. We thus aimed to establish through validation that cryopreserving and maintaining the iMSC-derived PDGFRα-positive cells for functional investigations help us to overcome these challenges. RESULTS: We demonstrated for the first time that the differentiated PDGFRα-positive cells from iMSCs can be cryopreserved and thawed to be used as per the experimental requirements with prolonged preservation of their characteristics. We assessed the viability of differentiated PDGFRα-positive cells pre- and post-freezing with the subsequent validation of their functional features using flow cytometry, qRT-PCR, and western blotting. We have been successful in demonstrating for the first time that the cryopreservation of previously differentiated PDGFRα-positive cells can be used as a feasible and cost-effective model for experimental reproducibility in functional studies of Diabetes Gastroparesis.

The Prevalence of Mild Cognitive Impairment (Mci) among Older Population of Rural Kerala: A Cross-Sectional Study.

Background: There are only very few studies on estimating the prevalence of mild cognitive impairment (MCI) from India, particularly from a rural setting. The available studies were heterogeneous. Objective: The study estimated the prevalence of MCI in a rural setting in Kerala, India. Materials and Methods: We conducted a community-based, cross-sectional study among individuals aged 65 and above in rural Thiruvananthapuram, Kerala. A cluster-randomized sampling was adopted, the cluster being the wards in the village. It was a two-phase door-to-door survey. Grassroots-level health workers enrolled 366 elders in the selected four wards in the initial phase and collected information on the sociodemographic details, comorbidities, and other risk factors of the participants, using a semi-structured questionnaire. Additionally, the Everyday Abilities Scale for India (EASI) was administered to assess their activities of daily living. In the second phase, a neurologist and a psychologist examined those screened positive with EASI and diagnosed MCI and dementia based on the MCI Working Group of the European Consortium on Alzheimer's Disease and the DSM V criteria, respectively. Results: The prevalence of MCI and dementia was 18.6% (95% confidence interval [CI] 14.7%-23.4%) and 6.8% (4.46%-10.1%), respectively, among the study participants. The prevalence of MCI was higher among the unemployed and those above 70 years of age. Conclusion: The community prevalence of MCI is more than three times that of dementia among the elderly in rural Kerala.

Normative Scores for Malayalam Cognitive Screening Test Battery among Older Adults in Kerala, India.

Background: Many neuropsychological tests are primarily developed in high-income countries, and normative data are not readily available for low- and middle-income countries (LMICs). We need culturally appropriate cognitive screening tests for India. Objective: Hence, we decided to translate the Hindi cognitive screening test battery (HCSTB) tool to Malayalam and establish the age and education-stratified norms for a Malayalam cognitive screening test battery (MCSTB). Material and Methods: HCSTB was translated to Malayalam, back-translated by bilinguals conversant in Malayalam and English, and pretested on 30 older normal adults. Using a multistage sampling technique, we conducted a descriptive cross-sectional survey in the Thiruvananthapuram district of Kerala, India. We approached older adults aged ≥60 years for informed and written consent. We excluded subjects with depression, functional impairment, cognitive impairment, history of stroke, psychosis, and visual/hearing loss that impaired cognitive assessment. Results: The normative data were derived from 441 older adults: 226 (51%) from rural areas and 215 (49%) from urban areas. Age and education affected the cognitive scores. The time to administer MCSTB among normal adults was approximately 17 minutes. Discussion and Conclusion: The derived normative data showed lower values than the published literature. A limitation of our study was the small number of older people with ≥12 years of education and the lack of neuroimaging of the subjects.

Neuropsychiatric Symptoms and Caregiver's Burden in Parkinson's Disease Patients in a Tertiary Care Teaching Hospital in South India: A Cross-Sectional Study.

BACKGROUND: In patients with Parkinson's disease (PD), the occurrence of motor and non-motor symptoms increases with disease progression. The range of neuropsychiatric symptoms (NPS) vary among individuals and can be burdensome for caregivers. Only a few studies have identified the contributing factors of NPS and caregiver burden in India. OBJECTIVES: We aimed to study the clinical profile, disability, and predictive factors of NPS in PD patients and associated caregiver's burden. METHODS AND MATERIAL: This was a cross-sectional observational study carried out in PD patients and their respective caregivers attending a movement disorder clinic in a tertiary care teaching hospital in Kerala. A total of 104 patients diagnosed with idiopathic PD receiving levodopa therapy and who had a primary caregiver were enrolled in the study. Structured questionnaires were administered to both patients and caregivers to collect data. Data analysis was done using an independent t-test, linear, and multiple regression analysis. RESULTS: Among 104 patients recruited for the study, 61.5% of patients had shown at least one NPS and 40.44% showed multiple NPS. Results from the study showed that depression is the primary NPS occurring in IPD patients (55.8%) followed by irritability, anxiety, and apathy. On linear regression models, the prime determinant of NPS was the Everyday Abilities Scale for India (EASI). For caregiver burden, the main determinants were the presence of NPS, duration of caregiving, EASI, and RBDSQ score. CONCLUSIONS: NPS in PD are highly associated with and are determinants of caregiver burden. Detailed assessment and specific interventions aimed at NPS could alleviate caregiver burden.

Predictors of Levo-dopa induced Dyskinesias in Parkinson's Disease.

BACKGROUND: Levodopa has a superior antiparkinsonian effect than dopamine agonists making it the standard of care for patients with Parkinson's disease (PD). During the initial stages, PD patients show a steady response to levodopa. Response fluctuations and levodopa-induced dyskinesias (LID) develop subsequently. The timing and onset of dyskinesias vary among individuals, and there are very few studies identifying the predictors of dyskinesia in India. AIMS: We aimed to study the clinical profile, disability, and predictors of LID in a patient with PD. MATERIALS AND METHODS: This was a cross-sectional observational study of consecutive patients with PD attending our movement disorder clinic. Patients on levodopa treatment with a minimum follow-up of 6 months were included in the study. All patients were observed before and after administration of levodopa to assess onset, duration of action, and timing of dyskinesias. Dyskinesias were video recorded and classified. Bivariate analysis was performed using Chi-square test or Fisher's exact test and multivariate analysis using binary logistic regression. RESULTS: This study recruited 110 patients with PD on levodopa therapy. Thirty-one (28.1%) out of 110 had LID. Of these, 25 patients (80.6%) had on-time dyskinesia, 19 patients (61.3%) had off-time dystonia, and 13 patients (41.9%) had diphasic dyskinesia. Majority had only mild-to-moderate dyskinesia. Incapacitating dyskinesias were during off time, primarily affecting the foot. Age, disease duration, disease severity, duration of treatment, and total dose of levodopa were found to be predictors of LID. Multivariate regression analysis showed younger age and longer duration of levodopa treatment to be independent predictors for LID. CONCLUSIONS: LID is fairly common in PD though not severely disabling. Patients with younger age of onset, longer disease duration, and severe disease were more likely to get early LID. We observed the lower prevalence of LID when initiating at lower doses and slow titration of levodopa.