Prediabetes in rural adolescent girls from DERVAN cohort: data from the KONKAN region of the state of Maharashtra, India (DERVAN-4).

Background: India is witnessing an epidemic of type 2 diabetes. Overweight/obesity, overnutrition, physical inactivity, and family history are well-known risk factors for diabetes. We investigated the role of undernutrition in the development of diabetes among rural adolescent girls. Methods: DERVAN cohort study was set up in the KONKAN region of the western Indian state of Maharashtra. It enrolled 1,520 adolescent girls (16-18 years old at the time of enrollment). We measured glycemic parameters (glucose, insulin, and HbA1C) and body size using anthropometry and body composition using bioimpedance. Prediabetes was diagnosed using the American Diabetic Association (ADA) criteria. We also calculated various HOMA indices for insulin resistance (HOMA-IR), ß-cell function (HOMA-ß), insulin sensitivity (HOMA-S), and compensatory ß-cell response using a homeostasis model. BMI, body fat%, and waist circumferences were treated as exposures and all the glycemic parameters and indices as outcomes. Results: The median age of the subjects was 16.6 years. The median weight, height, and BMI were 40.7 kg, 151.7 cm, and 17.5 kg/m2, respectively. Prevalence of underweight was 28.8%, and stunting was observed in 30.4%. Thinness and obesity using BMI were observed in 58.4% and 4.2%, respectively. The median body fat% was 22.5, and excess body fat (>35%) was observed in 5.7%. The prevalence of prediabetes was 39.4%. Fasting insulin concentrations, HOMA-IR, and HOMA-ß showed a positive trend across body composition quartiles (p < 0.001). HOMA-S and compensatory ß-cell response showed an inverse trend (p < 0.001). Compared with prediabetic girls in the overweight/obese group, girls most undernourished group had lower median insulin concentrations (8.1 µIU/ml vs. 17.1 µIU/ml), lower HOMA-IR (1.1 vs. 2.3), and lower HOMA-ß (75.6 vs. 129.2) but higher sensitivity (87.4 vs. 43.7) (p < 0.001) for all. Conclusion: We have reported a high prevalence of prediabetes among rural adolescent girls with a very low prevalence of obesity. Prediabetes in obesity is driven by hyperinsulinemia and overworking of the pancreas while poor ß-cell function and poor insulin secretion are major drivers in the undernourished group. The high-risk diabetes screening programs are much needed for the undernourished populations. Caution should be exercised for planning the interventions as overfeeding (or overnutrition) is likely to put the populations at risk of development of obesity and insulin resistance.

Diurnal variation in salivary progesterone in fertile Indian women.

Research question: Is there a diurnal variation in salivary progesterone levels during menstrual cycle among Indian women? Design: A longitudinal study was carried out to measure progesterone in saliva among small cross-sectional sample (n = 31) of fertile Indian women of reproductive age comprising young adults (18-25 years, n = 11), adults (26-38 years, n = 9) and middle aged (39-45 years, n = 11). Saliva samples were collected twice daily (morning and evening) across the entire menstrual cycle of 31 women. Results: Mean ages at enrolment and menarche were 30.6 years and 13.6 years respectively. Fifty-five percent of the women were married. The menstrual cycle range was 20-40 days. After controlling for age and menstrual cycle length, statistically significant diurnal variation in progesterone levels was observed across menstrual cycles with high levels in the morning. Conclusions: This is the first report on salivary progesterone in subjects with Indian ethnicity and could have clinical implications for designing point of care kits for menstrual cycle management, fertility and reproduction.

Determination of Manganese in Serum Using GFAAS: Serum Reference Values for Manganese in the Adolescent Girls of the DERVAN Cohort.

Current proposed method allows for the determination of manganese in serum sample using aqueous standard calibration technique on Graphite Furnace atomic absorption spectrophotometer (GFAAS) using deuterium background correction. This method involves determination of manganese from digested serum samples without the use of matrix modifier. Pyrolysis and atomization temperatures are 1200 °C and 2200 °C respectively. The limit of detection (LoD) and limit of quantitation (LoQ) of the method are 0.0097 ng/ml (0.18 nmol/l) and 0.032 ng/ml (0.58 nmol/l) respectively. Validation of the method was carried out using seronorm trace element level-1 serum standard with excellent agreement between measured value and certified value. Accuracy was demonstrated by the spike and recovery study with analytical recovery between 98.8 and 100.6% in serum. The serum reference value for manganese in adolescent girls of rural Konkan region of India range from 4.7 to 215 nmol/l.

aDolescent and prEconception health peRspectiVe of Adult Non-communicable diseases (DERVAN): protocol for rural prospective adolescent girls cohort study in Ratnagiri district of Konkan region of India (DERVAN-1).

INTRODUCTION: Precise impact of nutritional insufficiencies in adolescence as a risk factor for non-communicable diseases (NCD) in later life as adults remains largely unknown.We are conducting research into the effects of nutrition on adolescent girls of Ratnagiri district by a prospective cohort study (aDolescent and prEconception health peRspectiVe of Adult Non-communicable diseases cohort). Our study focuses on the physical health, nutritional parameters and cognitive profiles of adolescent girls, during the prenatal and postnatal period and we aim to follow this cohort and their offspring for 20 years. METHODS AND ANALYSIS: Cohort recruitment began in June 2019. Our aim is to recruit more than 1500 adolescent girls, age 16-18 years, over a period of 3 years. The recruit's cognition, diet and physical activity will be recorded. The following investigations will be performed: body composition by anthropometry and bioimpedence, and blood pressure, fasting blood sample to measure glucose, insulin, lipids, micronutrients and hormones, abdominal ultrasonography to measure liver, pancreas and kidneys.A biorepository has been created for long-term storage of blood, urine and saliva samples for future analysis. By this longitudinal study, we aim to identify the effects of malnutrition on the behavioural and biological measures in adolescent subjects and evaluate if these are associated with the onset of NCDs in adulthood. ETHICS AND DISSEMINATION: Institutional Ethic Committee (IEC) of BKL Walawalkar Rural Medical College and Hospital has granted the permission to carry out the study. IEC is registered with Government of India. Its registration code is EC/755/INST/MH/2015/RR-18. It is not a clinical trial but as required we have also registered the study on Clinical Trial Registry of India (CTRI). The registration code is CTRI/2019/04/018453.Appropriate written informed consent and assent are obtained from the parents and the adolescent girls, respectively. We plan to publish our results in peer-reviewed journals.

Expression, purification, and inhibition profile of dihydrofolate reductase from the filarial nematode Wuchereria bancrofti.

Filariasis is a tropical disease caused by the parasitic nematodes Wuchereria bancrofti and Brugia malayi. Known inhibitors of dihydrofolate reductase (DHFR) have been previously shown to kill Brugia malayi nematodes and to inhibit Brugia malayi DHFR (BmDHFR) at nanomolar concentrations. These data suggest that BmDHFR is a potential target for the treatment of filariasis. Here, protocols for cloning, expression and purification of Wuchereria bancrofti DHFR (WbDHFR) were developed. The Uniprot entry J9F199-1 predicts a 172 amino acid protein for WbDHFR but alignment of this sequence to the previously described BmDHFR shows that this WbDHFR sequence lacks a crucial, conserved 13 amino acid loop. The presence of the loop in WbDHFR is supported by a noncanonical splicing event and the loop sequence was therefore included in the gene design. Subsequently, the KM for dihydrofolate (3.7 ± 2 µM), kcat (7.4 ± 0.6 s-1), and pH dependence of activity were determined. IC50 values of methotrexate, trimethoprim, pyrimethamine, raltitrexed, aminopterin, (-)-epicatechin gallate, (-)-epicatechin, and vitexin were measured for WbDHFR and BmDHFR. Methotrexate and structurally related aminopterin were found to be effective inhibitors of WbDHFR, with an KI of 1.2 ± 0.2 nM and 2.1 ± 0.5 nM, respectively, suggesting that repurposing of known antifolate compound may be an effective strategy to treating filariasis. Most compounds showed similar inhibition profiles toward both enzymes, suggesting that the two enzymes have important similarities in their active site environments and can be targeted with the same compound, once a successful inhibitor is identified.

Discovery of a Stress-Activated Protein Kinase Inhibitor for Lymphatic Filariasis.

Lymphatic filariasis infects over 120 million people worldwide and can lead to significant disfigurement and disease. Resistance is emerging with current treatments, and these therapies have dose limiting adverse events; consequently new targets are needed. One approach to achieve this goal is inhibition of parasitic protein kinases involved in circumventing host defense mechanisms. This report describes structure-activity relationships leading to the identification of a potent, orally bioavailable stress activated protein kinase inhibitor that may be used to investigate this hypothesis.

Cognitive skills assessment during robot-assisted surgery: separating the wheat from the chaff.

OBJECTIVE: To investigate the utility of cognitive assessment during robot-assisted surgery (RAS) to define skills in terms of cognitive engagement, mental workload, and mental state; while objectively differentiating between novice and expert surgeons. SUBJECTS AND METHODS: In all, 10 surgeons with varying operative experience were assigned to beginner (BG), combined competent and proficient (CPG), and expert (EG) groups based on the Dreyfus model. The participants performed tasks for basic, intermediate and advanced skills on the da Vinci Surgical System. Participant performance was assessed using both tool-based and cognitive metrics. RESULTS: Tool-based metrics showed significant differences between the BG vs CPG and the BG vs EG, in basic skills. While performing intermediate skills, there were significant differences only on the instrument-to-instrument collisions between the BG vs CPG (2.0 vs 0.2, P = 0.028), and the BG vs EG (2.0 vs 0.1, P = 0.018). There were no significant differences between the CPG and EG for both basic and intermediate skills. However, using cognitive metrics, there were significant differences between all groups for the basic and intermediate skills. In advanced skills, there were no significant differences between the CPG and the EG except time (1116 vs 599.6 s), using tool-based metrics. However, cognitive metrics revealed significant differences between both groups. CONCLUSION: Cognitive assessment of surgeons may aid in defining levels of expertise performing complex surgical tasks once competence is achieved. Cognitive assessment may be used as an adjunct to the traditional methods for skill assessment during RAS.

Structure-activity studies of (-)-epigallocatechin gallate derivatives as HCV entry inhibitors.

Preventing viral entry into cells is a recognized approach for HIV therapy and has attracted attention for use against the hepatitis C virus (HCV). Recent reports described the activity of (-)-epigallocatechin gallate (EGCG) as an inhibitor of HCV entry with modest potency. EGCG is a polyphenolic natural product with a wide range of biological activity and unfavorable pharmaceutical properties. In an attempt to identify more drug-like EGCG derivatives with improved efficacy as HCV entry inhibitors, we initiated structure-activity investigations using semi-synthetic and synthetic EGCG analogs. The data show that there are multiple regions in the EGCG structure that contribute to activity. The gallate ester portion of the molecule appears to be of particular importance as a 3,4-difluoro analog of EGCG enhanced potency. This derivative and other active compounds were shown not to be cytotoxic in Huh-7 cell culture. These data suggest that more potent, non-cytotoxic EGCG analogs can be prepared in an attempt to identify more drug-like candidates to treat HCV infection by this mechanism.

Progress in the discovery and development of heat shock protein 90 (Hsp90) inhibitors.

The discovery and clinical development of heat shock protein 90 (Hsp90) inhibitors continue to progress. A number of Hsp90 inhibitors are in clinical trials, and preclinical discoveries of new chemotypes that bind to distinct regions in the protein as well as isoform selective compounds are active areas of research. This review will highlight progress in the field since 2010.

Discovery and optimization of 5-fluoro-4,6-dialkoxypyrimidine GPR119 agonists.

A series of 5-fluoro-4,6-dialkoxypyrimidine GPR119 modulators were discovered and optimized for in vitro agonist activity. A lead molecule was identified that has improved agonist efficacy relative to our clinical compound (APD597) and possesses reduced CYP2C9 inhibitory potential. This optimized lead was found to be efficacious in rodent models of glucose control both alone and in combination with a Dipeptidyl peptidase-4 (DPP-4) inhibitor.

Towards the discovery of drug-like epigallocatechin gallate analogs as Hsp90 inhibitors.

(-)-Epigallocatechin gallate (EGCG) is the major flavonoid of green tea and has been widely explored for a range of biological activities including anti-infective, anti-inflammatory, anti-cancer, and neuroprotection. Existing structure-activity data for EGCG has been largely limited to exploration of simple ethers and hydroxyl deletion. EGCG has poor drug-like properties because of multiple phenolic hydroxyl moieties and a metabolically labile ester. This work reports a substantial expansion of structure-activity understanding by exploring a range of semi-synthetic and synthetic derivatives with ester replacements and variously substituted aromatic and alicyclic groups containing more drug-like substituents. Structure-activity relationships for these molecules were obtained for Hsp90 inhibition. The results indicate that amide and sulfonamide linkers are suitable ester replacements. Hydroxylated aromatic rings and the cis-stereochemistry in EGCG are not essential for Hsp90 inhibition. Selected analogs in this series are more potent than EGCG in a luciferase refolding assay for Hsp90 activity.

Structure activity relationship study of benzo[d]thiazol-2(3H)one based σ receptor ligands.

Herein we report the SAR study which involved structural modifications to the linker length, aryl substitution and alkylamine ring size of the benzo[d]thiazol-2(3H)one based sigma receptor (σ) ligands. Many compounds in this series displayed low nanomolar affinity for the σ receptor subtypes. In particular, 8a showed high affinity (σ-1 Ki = 4.5 nM) for σ-1 receptors and moderately high selectivity (483-fold) over σ-2 receptors.

A 96-well filtration method for radioligand binding analysis of σ receptor ligands.

σ receptors represent a potential drug target for numerous therapeutic indications including cancer, depression, psychostimulant abuse, and stroke. Most published radioligand binding studies for σ receptors utilize a low throughput method employing a "cell harvester." Higher throughput methods are required to facilitate efficient screening of large numbers of novel compounds. In this study, a series of reference compounds was analyzed with a new medium-throughput 96-well filtration method and the results were compared to those obtained using the conventional cell harvester-based method. The 96-well assay utilized rat liver membranes for the determination of both known σ receptor subtypes (σ(1) and σ(2)) because this tissue contains high densities of both subtypes and fulfills criteria required for reliable use with the 96-well format. The new method gave comparable K(i) values for reference ligands analyzed in parallel with samples prepared in rat brain membranes and processed on the traditional cell harvester. For σ(1) receptors, equivalent affinity values were observed for both methods/tissues. For σ(2) receptors, approximately 2-fold higher affinities were observed for most compounds in liver, as compared to brain membranes, but excellent correlation with brain-derived values was maintained. To further demonstrate the utility of the new method it was used to screen a novel series of 2(3H)-benzothiazolone compounds, resulting in the identification of several analogues with nanomolar affinity and greater than 50-fold specificity for σ(1) versus σ(2) receptors.

Early development of sigma-receptor ligands.

Sigma receptors (σ-1 and σ-2) are non-opioid proteins implicated in the pathophysiology of various neurological disorders and cancer. The σ-1 subtype is a chaperon protein widely distributed in the CNS and peripheral tissues. These receptors are involved in the modulation of K(+)- and Ca(2+)-dependent signaling cascades at the endoplasmic reticulum and modulation of neurotransmitter release. σ-1 receptors are emerging targets for the treatment of neurophychiatric diseases (schizophrenia and depression) and cocaine addiction. σ-2 receptors are lipid raft proteins. They are highly expressed on many tumor cells and hence considered potential targets for anticancer drugs. σ receptors bind to a diverse class of pharmacological compounds like cocaine, methamphetamine, benzomorphans like (±)-pentazocine, (±)-SKF-10,047 and endogenous neurosteroids and sphingolipids. In this review we focus on the early development of σ receptor-specific ligands and radiolabeling agents.