Exposure assessment and risk characterisation of aflatoxins in randomly collected rice samples from local markets of Hyderabad, India.

Aflatoxin (AF) poisoning of staple foods, such as rice, is caused by fungal contamination by Aspergillus species. These AFs are genotoxic, carcinogenic and suppress the immune system. Hence, the present study was conducted to elucidate the prevalence of AF contamination in rice samples collected from local markets of Hyderabad, Telangana, India. The rice samples collected were analysed for AF by using HPLC-fluorescence detection (HPLC-FLD). Based on AF contamination levels and dietary intake of rice, the health risk was assessed by the margin of exposure (MOE) and liver cancer risk in adults, adolescence and children. The percentage detected contamination with AFB1 and AFB2 of rice samples was 54% and 34%, with the concentration ranging between 0-20.35 µg/kg and 0-1.54 µg/kg, respectively. Three rice samples exceeded the Food Safety and Standards Authority of India (FSSAI) total AF acceptable limit of 15 µg/kg. The average MOE values were 53.73, 50.58 and 35.69 (all <10,000) for adults, adolescence and children, respectively. The average liver cancer risk associated with rice consumption in the population of Hyderabad was found to be 0.27, 0.28 and 0.40 hepatocellular carcinoma (HCC) cases/year/100,000 individuals in adults, adolescence and children, respectively. This study revealed an adverse health risk to population of Hyderabad due to consumption of AF contaminated rice.

Chronic exposures to cholinesterase-inhibiting pesticides adversely affects the health of agricultural workers in India.

Biomonitoring of pesticide exposure has become a public concern because of its potential health effects. The present study investigated the acetylcholinesterase (AChE) inhibitory levels and their associated health effects in agricultural areas in Telangana, India. This cross-sectional included 341 exposed participants and 152 control participants from agricultural areas. A structured questionnaire was completed and blood and urine samples were collected to measure pesticides, dialkyle phosphate (DAP) metabolites, and AChE activity using liquid chromatography-tandem mass spectrometry and reversed-phase high-performance liquid chromatography. twenty-eight pesticides were detected in blood samples at concentrations ranging 0.42-45.77 ng/mL. Six DAP metabolites were also measured in urine, and all DAP metabolites were significantly higher in the exposed group. AChE activity is significantly reduced in individuals exposed for >10 years, raising concerns regarding possible neurological disorders. These results emphasise the urgent need to investigate the health effects of pesticides exposure, especially in agriculture.

Assessing farmer's exposure to pesticides and the risk for non-communicable diseases: A biomonitoring study.

This cross-sectional study aimed to assess the adverse effects of pesticide use in humans, such as non-communicable diseases, using acetylcholinesterase (AChE) and pesticide concentrations in blood samples. A total of 353 samples (290 case and 63 control) were collected from participants with >20 years of experience in agricultural pesticide use. The pesticide and AChE concentrations were determined using Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) and Reverse Phase High Performance Liquid Chromatography (RP-HPLC). Various health risks from pesticide exposure were assessed, including dizziness or headache, tension, anxiety, confusion, loss of appetite, loss of balance, concentration difficulties, irritability, anger, and depression. These risks may be influenced by the duration and intensity of exposure, the type of pesticide, and environmental factors in the affected areas. A total of 26 pesticides were detected in the blood samples from the exposed population, including 16 insecticides, three fungicides, and seven herbicides. Pesticide concentrations ranged from 0.20 to 12.12 ng/mL, and were statistically significant between the case and control groups (p < 0.05, p < 0.01, and p < 0.001). A correlation analysis was performed to determine statistically significance between pesticide concentration and symptoms of non-communicable diseases, such as Alzheimer's, Parkinson's, obesity, and diabetes. The estimated AChE levels in case and control blood samples were 21.58 ± 2.31 and 24.13 ± 1.08 U/mL, respectively (mean ± SD). The AChE levels were significantly lower in case samples than in controls (p < 0.001), which is considered to be an effect of long-term pesticide exposure, and is the cause of Alzheimer's disease (p < 0.001), Parkinson's disease (p < 0.001), and obesity (p < 0.01). Chronic exposure to pesticides and low levels of AChE are somewhat related to non-communicable diseases.

Sediment-water partitioning and risk assessment of organochlorine pesticides along the urban, peri-urban and rural transects of Krishna River Basin, Peninsular India.

Organochlorine pesticides (OCPs) were widely used in the past for pest control in agricultural lands and vector control programs in India. Due to their persistence and toxic impacts, we have quantified twenty OCPs in surface water, groundwater, and surface sediment samples along the Krishna River Basin (KRB), flowing through Peninsular India. Samples were collected along the urban, peri-urban, and rural transects of the KRB to understand the relation between the occurrence of pesticidal organochlorine pollutants based on the land use and land cover (LULC) and asses potential risk. Diagnostic ratios revealed ongoing Lindane usage in rural and peri-urban transects. On the contrary, the urban transect of the Musi River (MR) showed fresh inputs of technical HCH. The ratios of (p,p'-DDE+ p,p'-DDD)/ΣDDT >0.5 and α/ß-Endosulfan < 2.33 for most of the sites across the three transects for surface water, groundwater, and sediment indicate past DDT and Endosulfan usage across KRB. Excluding p,p'-DDE, and heptachlor in most of the sites, the logKOC' was higher than logKOC for other OCPs in the urban transect. However, for all the OCPs, the logKOC' was lower than logKOC in the peri-urban and rural transects of KRB thereby indicating that riverine sediment is acting as a sink for OCPs. The Krishna River annually transport about 0.24 tons HCH, 0.11 tons of DDT and 0.1 tons of Endosulfan. Despite having low water discharge, the compound-specific fluxes of the Wyra river are higher than the other two tributaries. Ecotoxicological risk assessment based on the Hazard Quotient suggested DDT pose higher risks to scud (zooplankton) and dinoflagellate and diatom (phytoplankton) whereas Endosulfan poses a threat to Bluegill (fish).

Ultra high-Performance Liquid Chromatography (UHPLC) method development and validation for the identification of oxidized product of Epigallocatechin-3-Gallate (EGCG).

OBJECTIVE: In the present study we reported oxidation of epigallocatechin-3-gallate (EGCG) and validation of oxidized product by a validated ultra high-performance liquid chromatography (UHPLC) method. METHODS: Two hundred milligrams of EGCG was oxidized in 5 mL of hydrogen peroxide (H2O2) and was identified by a validated UHPLC method with precision and robustness. Confirmation of parameters like C-H stretching and mass was carried out using infrared spectroscopy and mass spectroscopy, respectively. Identification of oxidized EGCG (O-EGCG) was done by UHPLC. RESULTS: The infrared spectroscopy chromatograms observed less intensity C-H stretching as compared to O-EGCG. The mass of EGCG and O-EGCG were 459.09 and 915.16, respectively. Structure elucidation revealed a loss of one proton in O-EGCG as compared to EGCG. Validation of the developed method was specific, with linear correlation coefficient 0.9981 and 0.9917, respectively for EGCG and O-EGCG, the accuracy rate of 95.2%-99.6% for EGCG, and 99.18%-101.5% for O-EGCG. CONCLUSION: Together, the results of this study demonstrate the formation of a dimer also the UHPLC method developed for identification of both EGCG and O-EGCG is validated as per the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines.

Placental mesenchymal stem cells restore glucose and energy homeostasis in obesogenic adipocytes.

Obesity (Ob) depicts a state of energy imbalance(s) being characterized by the accumulation of excessive fat and which predisposes to several metabolic diseases. Mesenchymal stem cells (MSCs) represent a promising option for addressing obesity and its associated metabolic co-morbidities. The present study aims at assessing the beneficial effects of human placental MSCs (P-MSCs) in mitigating Ob-associated insulin resistance (IR) and mitochondrial dysfunction both in vivo and in vitro. Under obesogenic milieu, adipocytes showed a significant reduction in glucose uptake, and impaired insulin signaling with decreased expression of UCP1 and PGC1α, suggestive of dysregulated non-shivering thermogenesis vis-a-vis mitochondrial biogenesis respectively. Furthermore, obesogenic adipocytes demonstrated impaired mitochondrial respiration and energy homeostasis evidenced by reduced oxygen consumption rate (OCR) and blunted ATP/NAD+/NADP+ production respectively. Interestingly, co-culturing adipocytes with P-MSCs activated PI3K-Akt signaling, improved glucose uptake, diminished ROS production, enhanced mitochondrial OCR, improved ATP/NAD+/NADP+ production, and promoted beiging of adipocytes evidenced by upregulated expression of PRDM16, UCP1, and PGC1α expression. In vivo, P-MSCs administration increased the peripheral blood glucose uptake and clearance, and improved insulin sensitivity and lipid profile with a coordinated increase in the ratio of ATP/ADP and NAD+ and NADP+ in the white adipose tissue (WAT), exemplified in WNIN/GR-Ob obese mutant rats. In line with in vitro findings, there was a significant reduction in adipocyte hypertrophy, increased mitochondrial staining, and thermogenesis. Our findings advocate for a therapeutic application of P-MSCs for improving glucose and energy homeostasis, i.e., probably restoring non-shivering thermogenesis towards obesity management.

Assessment of Antioxidant, Immunomodulatory Activity of Oxidised Epigallocatechin-3-Gallate (Green Tea Polyphenol) and Its Action on the Main Protease of SARS-CoV-2-An In Vitro and In Silico Approach.

Owing to the instability of Epigallocatechin Gallate (EGCG), it may undergo auto-oxidation and form oxidised products or dimers. In the present study, we aimed to evaluate the therapeutic effects, including antioxidation and immunomodulatory action, of the Oxidised Epigallocatechin Gallate (O-EGCG) as compared to native EGCG and the action of these compounds on main protease (Mpro) docking against SARS-CoV-2. HCT-116 (Human Colon Cancer) cell lines were used to estimate the total antioxidant capacity and lipid peroxidation levels and pro-inflammatory markers (human IL-6, IL-1ß, TNF-α). Further, molecular docking analysis was performed by AutoDock and visualised in Discovery studio. Improved antioxidant capacity of O-EGCG was observed, and there was a significant decrease in the inflammatory markers (IL-1ß, IL-6, and TNF-α) when O-EGCG was applied as compared to EGCG. The O-EGCG was shown to be strongly associated with the highest docking score and active site residues of IL-1, IL-6, and TNF- α, as well as the Mpro of SARS-CoV-2, according to in silico approach. The in vitro and in silico analyses indicate an improved therapeutic action of the oxidised form of EGCG. The effective inhibitory action of O-EGCG against SARS-CoV-2 suggests further exploration of the compound against COVID-19 and its efficacy. However, in vivo studies and understanding of the mechanism of action of O-EGCG may yield a better opinion on the use of O-EGCG and future human clinical trials.

Human placental mesenchymal stromal cell therapy restores the cytokine efflux and insulin signaling in the skeletal muscle of obesity-induced type 2 diabetes rat model.

Obesity poses a significant risk factor for the onset of metabolic syndrome with allied complications, wherein mesenchymal stem cell therapy is seen as a promising treatment for obesity-induced metabolic syndrome. In the present study, we aim to explore the beneficial effects of the human placental mesenchymal stromal cells (P-MSCs) on obesity-associated insulin resistance (IR) including inflammation. To understand this, we have analyzed the peripheral blood glucose, serum insulin levels by ELISA, and the glucose uptake capacity of skeletal muscle by a 2-NBDG assay using flow cytometry in WNIN/GR-Ob rats treated with and without P-MSCs. Also, we have studied insulin signaling and cytokine profile in the skeletal muscle by western blotting, dot blotting, and Multiplex-ELISA techniques. The skeletal muscle of WNIN/GR-Ob rats demonstrates dysregulation of cytokines, altered glucose uptake vis-a-vis insulin signaling. However, P-MSCs' treatment was effective in WNIN/GR-Ob rats as compared to its control, to restore HOMA-IR, re-establishes dysregulated cytokines and PI3K-Akt pathway in addition to enhanced Glut4 expression and glucose uptake studied in skeletal muscle. Overall, our data advocate the beneficial effects of P-MSCs to ameliorate inflammatory milieu, improve insulin sensitivity, and normalize glucose homeostasis underlining the Ob-T2D conditions, and we attribute for immunomodulatory, paracrine, autocrine, and multipotent functions of P-MSCs.

Understanding the Alterations in Lipid Metabolism in NAFLD Progression: Current Trends and Future Directions.

Nonalcoholic fatty liver disease (NAFLD) is a pathological condition, wherein fat deposition exceeds the allowable limits of the healthy person. If the condition persists for a long time, the patient will eventually develop NASH (nonalcoholic steatohepatitis), which will probably lead to HCC (hepatocellular carcinoma). The incidence of NAFLD is rising at an alarming rate, and still there are no drugs approved by the U.S. Food and Drug Administration for this devastating health condition. To combat and treat NAFLD successfully, it is essential to understand how routine lipid metabolism in the liver is altered under these conditions. In this review, we discuss specifically during the NAFLD progression how the signaling pathways leading to excess fat accumulation in the liver are changed. We also address variations in the mechanisms underlying hepatic lipid uptake and changes in fatty acid oxidation mechanisms. We will also highlight the role of transcription factors and other lipolytic enzymes that stringently regulate the hepatic de novo lipolysis (DNL) and emphasize how they are altered during NAFLD progression. Finally, we will also touch upon how the lipid disposal from the liver goes wrong during the NAFLD progression. A comprehensive understanding of the changes in lipid metabolism is essential for developing successful therapies for NAFLD.

Toxicokinetic analysis of commonly used pesticides using data on acute poisoning cases from Hyderabad, South India.

Organophosphorus pesticides (OPs) interfere with the activity of acetylcholinesterase (AChE), a vital enzyme that regulates the functioning of the nervous system, resulting in acetylcholine (Ach) accumulation at the synapses and myoneural junctions. It remains unknown whether the commonly used OPs in South India also interfere with the AChE activity and their toxicokinetics in humans remains poorly understood. We collected peripheral blood samples from OP-associated suicide cases (hospitalised) and analysed the pesticide concentration and AChE activity, and the toxicokinetics of six commonly used pesticides. LC-MS/MS was used for the estimation of pesticide concentration. Based on a comparison of six pesticide kinetic profiles and toxicokinetic parameters, we concluded that chlorpyrifos ingestion resulted in the highest concentration of chlopyrifos among the identified pesticides, followed by acephate, triazophos, propanil, while dimethoate exhibited the lowest concentration. Based on a time-course analysis, we observed a faster elimination phase for monocrotophos and dimethoate. We observed that there was a significant decrease in the mean concentration of monocrotophos (64 ng/mL) (P = 0.015), while the mean value of AChE (1.08 unit/mL) increased over time. While monocrotophos and dimethoate elimination phases were remarkable in human subjects, the other pesticides did notdemonstrate similar elimination phases owing to their low rate of metabolism and high stability.

Accumulation of polychlorinated biphenyls in fish and assessment of dietary exposure: a study in Hyderabad City, India.

The objective of this study is to determine the concentrations of polychlorinated biphenyls (PCBs) in different fish species collected from fish outlets in the south Indian city of Hyderabad. The samples of fish extracted by using Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) and concentrations of polychlorinated biphenyls (PCBs, 45 congeners) were determined by gas chromatography-microelectron capture detection (GC-µECD) method. The intake of individual PCB congeners through fish, toxicity equivalence factors (TEFs), hazardous quotient, quantitative assessment, and risk evaluations was estimated in the Indian population. Daily dietary intakes of PCBs at the 95th-percentile-measured concentrations were twice the values of the 50th-percentile-measured concentrations in all socio-economic groups. The dietary intakes of PCBs through fish consumption in middle-income group, low-income group, and industrial laborers (0.023 µg kg(-1) day(-1)), the high-income group (0.031 µg kg(-1) day(-1)), and slum dwellers (0.039 µg kg(-1) day(-1)) exceeded the reference dose. The observed estimated daily intakes (EDIs) of our study for all socio-economic sections (0.0087 µg/kg/day) crossed the cancer benchmark concentration of 0.0003 µg/kg/day. In slum dwellers, the ingestion of fish from freshwater and marine water results in the highest lifetime cancer risks of 4.7 in 100,000 and 7.8 in 100,000, respectively. Ultimately, the concentrations of PCBs were determined high in all of the fish species collected. Risk assessment showed that the fish were highly contaminated with PCBs and may pose health threats to consumers in the city of Hyderabad as well as a lifetime cancer risk.

New mathematical derivations for calculation of ATP yield due to the complete oxidation of different types of fatty acids.

During the complete oxidation of fatty acids, the electrons removed from fatty acids in different forms (FADH2 and NADH2) pass through the respiratory chain, driving the ATP synthesis. Generally, the ATP yield due to the complete oxidation of fatty acids is calculated by sum total the ATPs obtained due to the oxidation of FADH2 and NADH2 due to lack of any particular method. This calculation is simple for saturated even numbered fatty acids, but in the case of saturated and unsaturated odd numbered fatty acids the calculation of ATP yield is difficult and needs mathematical calculations due to some changes in their beta-oxidation pathway when compared to the pathway of saturated even numbered fatty acids. These calculations are made simple by our derivations and following formulae where we require only number of carbon atoms and double bonds present in a fatty acid. Our method is superior and easier in comparison to long mathematical calculations that are in the practice.

Environmental monitoring of benzene and toluene produced in indoor air due to combustion of solid biomass fuels.

Exposure to benzene and toluene from the combustion of solid biomass fuels is one of the important causes of morbidity and mortality in developing countries. In this study, we assessed the exposure of cooks to benzene and toluene from biomass fuel combustion in 55 rural homes. The GC-MS was used for quantification while a personnel sampler was used for environmental monitoring. The benzene exposure differed significantly (p < 0.0001) across different types of indoor kitchen fuel combinations. The geometrical mean (GM) of benzene exposure for cooks during cooking hours in an indoor kitchen using mixed fuel was 75.3 microg/m3 (with partition) and 63.206 microg/m3 (without partition), while the exposure was 11.7 microg/m3 for open type. The benzene exposure was significantly higher (p < 0.05) in an indoor kitchen with respect to open type using mixed fuels. Concentration of benzene (114.1 microg/m3) for cooks in an indoor kitchen with partition using dung fuel was significantly higher in comparison to non-cooks (5.1 microg/m3) for open type. Benzene exposure was not significantly different for kitchen with ventilation (31.2 microg/m3) and without ventilation (45.0 microg/m3) using wood fuel. However, this value was significantly (p < 0.05) lower than in indoor kitchens with or without partition. An almost similar trend was observed for toluene but the difference was statistically non-significant. This study may be helpful in developing a regional exposure database and in the facilitation of health risk assessment due to volatile organic pollutants in our day-to-day environment.

Gas chromatographic-mass spectroscopic determination of benzene in indoor air during the use of biomass fuels in cooking time.

A gas chromatography-mass spectroscopic method in electron ionization (EI) mode with MS/MS ion preparation using helium at flow rate 1 ml min(-1) as carrier gas on DB-5 capillary column (30 m x 0.25 mm i.d. film thickness 0.25 microm) has been developed for the determination of benzene in indoor air. The detection limit for benzene was 0.002 microg ml(-1) with S/N: 4 (S: 66, N: 14). The benzene concentration for cooks during cooking time in indoor kitchen using dung fuel was 114.1 microg m(-3) while it was 6.6 microg m(-3) for open type kitchen. The benzene concentration was significantly higher (p < 0.01) in indoor kitchen with respect to open type kitchen using dung fuels. The wood fuel produces 36.5 microg m(-3) of benzene in indoor kitchen. The concentration of benzene in indoor kitchen using wood fuel was significantly (p < 0.01) lower in comparison to dung fuel. This method may be helpful for environmental analytical chemist dealing with GC-MS in confirmation and quantification of benzene in environmental samples with health risk exposure assessment.