Endothelin-1 mediated vasoconstriction leads to memory impairment and synaptic dysfunction.

Cerebrovascular lesions seen as white matter hyperintensity in MRI of elderly population caused due to micro-infracts and micro-bleeds contributes to vascular dementia. Such vascular insult caused by impairment in blood flow to specific area in brain involving small vessels can gradually worsen the pathology leading to cognitive deficits. In the present study we developed a transient model of vaso-constriction to study the impact of such pathology by bilateral injection of ET-1 (Endothelin-1; a 21 amino acid vasoconstricting peptide) into lateral ventricles of C57 mice. The impediment in cerebral blood flow decreased CD31 expression in endothelial cells lining the blood vessels around the hippocampal region, leading to memory deficits after 7 days. Activity dependent protein translation, critical for synaptic plasticity was absent in synaptoneurosomes prepared from hippocampal tissue. Further, Akt1- mTOR signaling cascade was downregulated indicating the possible cause for loss of activity dependent protein translation. However, these effects were reversed after 30 days indicating the ephemeral nature of deficits following a single vascular insult. Present study demonstrates that vasoconstriction leading to memory deficit and decline in activity dependent protein translation in hippocampus as a potential molecular mechanism impacting synaptic plasticity.

SARS-CoV2 genome analysis of Indian isolates and molecular modelling of D614G mutated spike protein with TMPRSS2 depicted its enhanced interaction and virus infectivity

COVID-19 that emerged as a global pandemic is caused by SARS-CoV-2 virus. The virus genome analysis during disease spread reveals about its evolution and transmission. We did whole genome sequencing of 225 clinical strains from the state of Odisha in eastern India using ARTIC protocol-based amplicon sequencing. Phylogenetic analysis identified the presence of all five reported clades 19A, 19B, 20A, 20B and 20C in the population. The analyses revealed two major routes for the introduction of the disease in India i.e. Europe and South-east Asia followed by local transmission. Interestingly, 19B clade was found to be much more prevalent in our sequenced genomes (17%) as compared to other genomes reported so far from India. The haplogroup analysis for clades showed evolution of 19A and 19B in parallel whereas the 20B and 20C appeared to evolve from 20A. Majority of the 19A and 19B clades were present in cases that migrated from Gujarat state in India suggesting it to be one of the major initial points of disease transmission in India during month of March and April. We found that with the time 20A and 20B clades evolved drastically that originated from central Europe. At the same time, it has been observed that 20A and 20B clades depicted selection of four common mutations i.e. 241 C>T (5UTR), P323L in RdRP, F942F in NSP3 and D614G in the spike protein. We found an increase in the concordance of G614 mutation evolution with the viral load in clinical samples as evident from decreased Ct value of spike and Orf1ab gene in qPCR. Molecular modelling and docking analysis identified that D614G mutation enhanced interaction of spike with TMPRSS2 protease, which could impact the shedding of S1 domain and infectivity of the virus in host cells.

Glutaredoxin1 Diminishes Amyloid Beta-Mediated Oxidation of F-Actin and Reverses Cognitive Deficits in an Alzheimer's Disease Mouse Model.

Aims: Reactive oxygen species (ROS) generated during Alzheimer's disease (AD) pathogenesis through multiple sources are implicated in synaptic pathology observed in the disease. We have previously shown F-actin disassembly in dendritic spines in early AD (34). The actin cytoskeleton can be oxidatively modified resulting in altered F-actin dynamics. Therefore, we investigated whether disruption of redox signaling could contribute to actin network disassembly and downstream effects in the amyloid precursor protein/presenilin-1 double transgenic (APP/PS1) mouse model of AD. Results: Synaptosomal preparations from 1-month-old APP/PS1 mice showed an increase in ROS levels, coupled with a decrease in the reduced form of F-actin and increase in glutathionylated synaptosomal actin. Furthermore, synaptic glutaredoxin 1 (Grx1) and thioredoxin levels were found to be lowered. Overexpressing Grx1 in the brains of these mice not only reversed F-actin loss seen in APP/PS1 mice but also restored memory recall after contextual fear conditioning. F-actin levels and F-actin nanoarchitecture in spines were also stabilized by Grx1 overexpression in APP/PS1 primary cortical neurons, indicating that glutathionylation of F-actin is a critical event in early pathogenesis of AD, which leads to spine loss. Innovation: Loss of thiol/disulfide oxidoreductases in the synapse along with increase in ROS can render F-actin nanoarchitecture susceptible to oxidative modifications in AD. Conclusions: Our findings provide novel evidence that altered redox signaling in the form of S-glutathionylation and reduced Grx1 levels can lead to synaptic dysfunction during AD pathogenesis by directly disrupting the F-actin nanoarchitecture in spines. Increasing Grx1 levels is a potential target for novel disease-modifying therapies for AD.

Cutaneous leishmaniasis in Bikaner, India: Clinicoepidemiological profile; parasite identification using conventional, molecular methods and CL Detect™ rapid test, a new Food and Drug Administration-approved test.

BACKGROUND AND OBJECTIVE: Cutaneous Leishmaniasis (CL) is a dermal manifestation caused by various species of Leishmania. This is the most common Leishmanial syndrome seen worldwide and is emerging as and threatens to become an uncontrollable disease. The present study was planned to understand the current epidemiology of CL in the conventionally endemic area of Bikaner, Rajasthan. Species characterization was also done. MATERIAL AND METHODS: The diagnostic modalities used in the study were microscopy, histopathologic examination, antigen testing and confirmation and species characterization by PCR. Field evaluation of the new antigen detection kit approved by FDA in 2016 CL Detect™ IC-RDT was done and its results compared with other available diagnostic tests. RESULTS: A total of 14 cases with 25 skin lesions presented to the Skin OPD during the six month study period (January to June 2018). Out of these, 5 were males and 9 were females, average age being 32 yrs. Students and housewives of poor socio economic group were the most frequently affected groups. Lesions were found on the exposed areas of the body, mainly on the upper limbs and facial region. Majority of them were ulcerated plaques. Out of 14 cases under study, antigen test confirmed 7 positives; thereby yielding a positive rate of 50% against 71.4% for microscopy, 72.7% for PCR and a mere 38.5% for histopathology. Species characterization revealed Leishmania tropica as the causative organism. INTERPRETATION AND CONCLUSION: The study indicates that the number of cases have declined substantially over the past decade in Bikaner. Despite being the oldest technique used, microscopy still is a good option for diagnosis. Combined with PCR, diagnostic accuracy and hence utility is increased multifold. In its current form, IC-RDT may not be very useful.

Discovery and preclinical development of IIIM-160, a Bergenia ciliata-based anti-inflammatory and anti-arthritic botanical drug candidate / 中西医结合学报

OBJECTIVE@#Bergenia ciliata (Haw.) Sternb. is used in the Indian traditional system of medicine to treat various ailments including rheumatism and to heal wounds. The objective of the present study was to perform a preclinical characterization of the B. ciliata-based botanical extract IIIM-160.@*METHODS@#IIIM-160 was chemically standardized and analyzed for heavy metal content, aflatoxins, pesticides and microbial load. The in vitro and in vivo efficacies were determined in suitable models of inflammation, arthritis and nociception. An acute oral toxicity study was performed in Swiss albino mice. A suitable oral formulation was developed and characterized.@*RESULTS@#Bergenin was found to be the major component (9.1% w/w) of IIIM-160. The botanical lead displayed inhibition of lipopolysaccharide-induced production of proinflammatory cytokines in THP-1 cells, with selectivity toward interleukin-6 (IL-6) and had an excellent safety-window. It showed anti-inflammatory, anti-arthritic and antinociceptive activity in animal models and was not toxic at oral doses up to 2 g/kg in Swiss-albino mice. The gastroretentive, sustained-release capsule formulation showed sustained-release of the bergenin over the period of 24 h, resulting in improved plasma-exposure of bergenin in Sprague-Dawley rats.@*CONCLUSION@#The dual-activity of IL-6 inhibition and antinociception marks the suitability of IIIM-160 for treating rheumatoid arthritis. This study will serve as the benchmark for further research on this botanical formulation.

A review of trade practices of smokeless tobacco products in terms of prohibition on sale, manufacturing & importation in Framework Convention on Tobacco Control ratified Parties.

BACKGROUND & OBJECTIVES: Over the past decade, the World Health Organization (WHO) Framework Convention on Tobacco Control (FCTC) has served as a powerful tool to initiate and advance global tobacco control efforts. However, the control strategies have mainly targeted demand-side measures. The goal of a tobacco-free world by 2040 cannot be achieved if the supply-side measures are not addressed. This analysis was undertaken to examine the tobacco control legislations of various Parties ratifying WHO FCTC with an objective to ascertain the status of prohibition of importation, sale and manufacturing of smokeless tobacco products. METHODS: All 180 Parties to WHO FCTC were included for the study. A comprehensive database of all the parties to FCTC was created and tobacco control legislations and regulations of all parties were studied in detail. RESULTS: Overall, the sale of smokeless tobacco (SLT) products was prohibited in 45 Parties. Eleven Parties prohibited manufacturing of SLT products and six Parties imposed a ban on importation of SLT products. Australia, Bhutan, Singapore and Sri Lanka banned all three. INTERPRETATION & CONCLUSIONS: Comprehensive tobacco control strategy with effective tobacco cessation programme should complement strong legal actions such as prohibition on trade in SLT products to meet the public health objective of such laws and regulations. In addition, multisectoral efforts are needed for effective implementation of such restrictions imposed by the governments.

Lessons for addressing noncommunicable diseases within a primary health-care system from the Ballabgarh project, India

Background: Most patients with noncommunicable diseases (NCDs) can bemanaged appropriately at the primary care level, using a simplified standardprotocol supported by low-cost drugs. The primary care response to commonNCDs is often unstructured and inadequate in low- and middle-income countries.This study assessed the feasibility of integration of NCD prevention and controlwithin the primary health-care system of India.Methods: This study was done among 12 subcentres, 2 primary health centres(PHCs) and one subdistrict hospital in a block in north India. All 28 multipurposehealth workers of these subcentres underwent 3-day training for delivering thepackage of NCD interventions as a part of their routine functioning. A time–motionstudy was conducted before and after this, to assess the workload on a sample ofthe workers with and without the NCD work. Screening for risk assessment wasdone at domiciliary level as well as at health-facility level (opportunistic screening),and the cost was estimated based on standard costing procedures. Individualswho screened positive were investigated with electrocardiography and fastingblood sugar. PHCs were strengthened with provision of essential medicines andtechnologies.Results: After training, 6% of the time of workers (n = 7) was spent in the NCDrelated activities, and introduction of NCD activities did not impact the coverage ofother major national health programmes. Loss during referral of “at-risk” subjects(37.5% from home to subcentre and 33% from subcentre to PHC) resulted inscreening efficiency being lowest at domiciliary level (1.3 cases of NCDs identifiedper 1000 screened). In comparison to domiciliary screening (`21 830.6; US$ 363.8per case identified), opportunistic screening at subdistrict level (`794.6; US$ 13.2)was 27.5 times and opportunistic screening at PHC (`1457.5; US$ 24.3) was 15.0times lower. There was significant utilization of NCD services provided at PHCs,including counselling.Conclusion: Opportunistic screening appears to be feasible and a cost-effectivestrategy for risk screening. It is possible to integrate NCD prevention and controlinto primary health care in India.

Effectiveness of a school based intervention for prevention of non-communicable diseases in middle school children of rural North India: a randomized controlled trial.

OBJECTIVE: To assess the effectiveness of a multi-component school based intervention in improving knowledge and behavioral practices regarding diet, physical activity and tobacco use in middle schoolchildren of rural-Ballabgarh, North-India. METHODS: A total of 40 middle schools were grouped into two, based on geographic proximity and randomly assigned to the intervention or control group in a cluster randomized controlled trial. The target population consisted of 2,348 children studying in 6th and 7th grades in these schools. The intervention consisted of a school component (policies), a classroom component (activities) and a family component [Information Education & Communication (IEC) material]. The main outcome measures were knowledge and behavioral changes in physical activity, diet and tobacco which were self- reported. RESULTS: Post-intervention, a significant number of intervention schools adopted the tobacco policy (16/19), physical activity policy (6/19) and healthy food policy (14/19) as compared to the control schools (n = 21). Knowledge about physical activity, diet and tobacco improved significantly in the intervention group as compared to the control group. Proportion of students attending Physical Training (PT) classes for five or more days in a week in the intervention group compared to the control group increased significantly (17.8%; p < 0.01). Proportion of students consuming fruits increased in the intervention group compared to the control group (10%; p < 0.01). Pre-post decrease in the prevalence of current smoking was significantly more in the intervention group as compared to the control group (7.7%; p < 0.01). CONCLUSIONS: Healthy settings approach for schools is feasible and effective in improving knowledge and behavioral practices of non-communicable diseases (NCD) risk factors in adolescents in rural India.

Lessons for addressing noncommunicable diseases within a primary health-care system from the Ballabgarh project, India.

BACKGROUND: Most patients with noncommunicable diseases (NCDs) can be managed appropriately at the primary care level, using a simplified standard protocol supported by low-cost drugs. The primary care response to common NCDs is often unstructured and inadequate in low- and middle-income countries. This study assessed the feasibility of integration of NCD prevention and control within the primary health-care system of India. METHODS: This study was done among 12 subcentres, 2 primary health centres (PHCs) and one subdistrict hospital in a block in north India. All 28 multipurpose health workers of these subcentres underwent 3-day training for delivering the package of NCD interventions as a part of their routine functioning. A time-motion study was conducted before and after this, to assess the workload on a sample of the workers with and without the NCD work. Screening for risk assessment was done at domiciliary level as well as at health-facility level (opportunistic screening), and the cost was estimated based on standard costing procedures. Individuals who screened positive were investigated with electrocardiography and fasting blood sugar. PHCs were strengthened with provision of essential medicines and technologies. RESULTS: After training, 6% of the time of workers (n = 7) was spent in the NCD-related activities, and introduction of NCD activities did not impact the coverage of other major national health programmes. Loss during referral of "at-risk" subjects (37.5% from home to subcentre and 33% from subcentre to PHC) resulted in screening efficiency being lowest at domiciliary level (1.3 cases of NCDs identified per 1000 screened). In comparison to domiciliary screening (₹21 830.6; US$ 363.8 per case identified), opportunistic screening at subdistrict level (₹794.6; US$ 13.2) was 27.5 times and opportunistic screening at PHC (₹1457.5; US$ 24.3) was 15.0 times lower. There was significant utilization of NCD services provided at PHCs, including counselling. CONCLUSION: Opportunistic screening appears to be feasible and a cost-effective strategy for risk screening. It is possible to integrate NCD prevention and control into primary health care in India.