Genetic Polymorphism in Xenobiotic Metabolising Genes and Increased Oxidative Stress among Pesticides Exposed Agricultural Workers Diagnosed with Cancers.

BACKGROUND: Oxidative stress combined with nullity of xenobiotic metabolizing GSTT1/GSTM1/CYP2E1 genes may increase the susceptibility of agricultural workers to adverse health effects including cancer. The present study was conducted to determine; the prevalence of polymorphisms in GSTM1, GSTT1 and CYP2E1 genes, serum 8-hydroxy-2'-deoxygunosine levels, and the role of these markers in risk of cancer among agricultural workers occupationally exposed to pesticides. METHODS: A total of 360 participants, of which 180 belonging to farming group diagnosed with leukemia (n=60), lymphoma (n=60) and breast cancers (n=60), 90 in non-farming group diagnosed with similar cancers and the other 90 as healthy controls with neither history of occupational exposure nor diagnosed with any type of cancers were recruited. Following the questionnaire survey, serum 8-OHdG and genetic polymorphisms in the three genes were determined using ELISA and PCR methods respectively. RESULTS: The results of the study revealed that farm workers carrying GSTT1 null genotype had increased risk for lymphoma (OR = 5.34; 95% CI = 1.80-15.82) and breast cancer (OR=4.04; 95% CI = 1.24-13.07). For farm workers carrying GSTM1 null genotype, the risk was six-fold for breast cancer (OR = 6.88; 95% CI =1.88-25.99). Further, there found a significant difference between 8-OHdG and nullity of CYP2E1 among the farm workers diagnosed with leukemia. CONCLUSION: The findings of the present study suggest that the polymorphisms in detoxifying genes among farm workers occupationally exposed to pesticides and the oxidative stress may likely be responsible for triggering the mechanism of malignancy.

Mitigation of pesticide residue levels in the exposed dermal regions of occupationally exposed farmworkers by use of personal protective equipment.

Unsafe pesticide handling practices with the limited use of personal protective equipment (PPE) by the Indian farming groups lead to an increased risk of exposure to pesticides. Therefore, a community-based follow-up study based on dosimeters, wipes, and hand-wash technique was carried out to evaluate the dermal exposure to pesticides and to analyze the impact of the usage of PPE on minimizing the exposure among the farmworkers of Rangareddy district, Telangana, India. Risk in terms of hazard quotient (HQ), hazard index (HI), and safety analysis as margins of safety was assessed. Farmworkers averaged 18 years of farming experience and showed resistance to adopting good agricultural practices. Ten pesticide residues were detected in concentrations ranging from 0.000 to 246 mg ml-1 in hand-wash, 0.000 to 198.33 ng cm-2 in patch dosimeter, and 0.000 to 1,740 ng cm-2 in wipe samples collected from farmworkers not using PPE. The second phase includes the intervention study results that revealed a significant reduction both in the concentrations and the number of pesticide residues detected in the hand-wash, patch, and wipe samples of the farmworkers who have used the PPE provided to them (p < 0.01). Furthermore, the probabilistic health risk assessment in terms of the HQ values ranged from 0.02 to 1029.82, and HI was >1, suggesting the non-carcinogenic risks associated with dermal exposure to pesticides among them. Additionally, the safety risk assessment in terms of the margin of safety suggests that they follow risky handling practices. The study confirms that farmworkers are exposed to pesticides and emphasizes the significance of using PPE in reducing the risk.

Residual Dialkyl Phosphate Metabolite Concentrations of Organophosphate Pesticides Among Indian Farmworkers: Implication of Exposure and Hazard Assessment.

OBJECTIVE: Biomonitoring of urinary dialkyl phosphate (DAP) metabolites, a sensitive biomarker to assess pesticides exposure and also to study the impact of the use of personal protective equipment (PPE). METHODS: A preintervention-postintervention study to biomonitor dimethylphosphate, diethylphosphate, diethylthiophosphate, and diethyl-dithiophosphate using liquid chromatography with tandem mass spectrometry among Indian farmworkers (n = 120). RESULTS: Dimethylphosphate was detected in all samples at a mean concentration of 74.91 µg · L -1 (17.616 µg · g -1 creatinine), whereas diethylthiophosphate and diethyl-dithiophosphate were detected in 88% and 82% of samples, respectively, among farmworkers who adopted unsafe pesticide-handling practices. Intervention studies showed a significant reduction in the urinary DAP metabolites detected among the farmworkers using PPE provided to them ( P < 0.01). CONCLUSIONS: Study confirms the exposure to pesticides among farmworkers and highlights the importance of the use of PPE to minimize exposure.

Knowledge, attitude and practices related to pesticide usage among farmers: Findings from a hospital based cross-sectional study.

BACKGROUND: In India, where agriculture is the prime occupation among the rural population, farmers are extensively using pesticides in crop production without taking adequate precautions. OBJECTIVE: The current study aimed to examine pesticide usage patterns, usage of personal protective equipment (PPE), knowledge on pesticide toxicity, and attitude towards occupational safety among farmers diagnosed with leukaemia (n = 60), lymphoma (n = 33), and breast cancer (n = 30) visiting a tertiary cancer care hospital in Hyderabad, India. METHODS: Face-to-face interviews were conducted using a pre-tested questionnaire and the data collected were analysed with statistical software SPSS 23. RESULTS: The survey revealed that the participants had an average of 19.6 years of farming experience and women were predominantly involved in the mixing of pesticides and other agricultural work apart from spraying. All of them were not using any PPE and the majority of them did not have any knowledge on the route of exposure, toxicity symbols, and safe handling of pesticides. Statistical analysis showed no significant association between educational status and knowledge, attitude and practices (p > 0.05). CONCLUSION: Even though the majority of the farmers were educated, their lack of adequate knowledge on pesticides had enabled them to adopt risky behaviours in handling and storage of pesticides. There is a need for continuous training programs for farmers to promote awareness and minimize the hazards of occupational exposure.

The impact of the use of personal-protective-equipment on the minimization of effects of exposure to pesticides among farm-workers in India.

Introduction: Although excessive use of pesticides and unsafe agricultural practices may contribute to numerous intoxications, the role of PPE (personal-protective-equipment) in the minimization of toxicological effects due to pesticide exposure has not been addressed so far. The present study aimed to assess the impact of the use of PPE on the minimization of effects of exposure to pesticides among farm-workers. Methods: A community-based follow-up study with questionnaire-based survey and field observations was undertaken among farm-workers (n = 180) of Rangareddy district, Telangana, India. Biomarkers of exposure such as cholinesterase activity, inflammatory markers (TNF-α, IL-1ß, IL-6, cortisol, and hs-C reactive protein), nutrients (vitamins A, E), liver function (total protein and A/G ratio, AST and ALT levels) were investigated in the laboratory by following the standard protocols. Results: Farm-workers who had a mean farming exposure of 18 years of and who neither followed safe pesticide handling practices nor used PPE and also showed reluctance to obey good agricultural practices (GAPs). Inhibition of AChE (acetylcholine esterase) with increased inflammation was found among farm-workers as compared to their respective normal values when they have not used PPE. Linear regression statistical analysis revealed a profound effect on inhibition in the AChE activity and various inflammatory markers with the increase in the duration of pesticide exposure. Further, there was no effect of the duration of pesticide exposure on the levels of vitamins A, E, ALT, AST, total protein, and A/G ratio. Further, intervention studies carried out on the use of PPE provided (commercially available and cost-effective) for 90 days showed a significant reduction in the biomarker levels (p < 0.01). Conclusion: This study demonstrated the importance of the use of PPE during pesticide applications and other agricultural tasks to minimize pesticide-associated adverse health effects.

Assessment of dermal exposure to pesticides among farmers using dosimeter and hand washing methods.

Inappropriate use of pesticides followed by unsafe handling practices to control the insect infestation among the farming groups in developing countries has resulted in a high exposure risk. The use of personal protective equipment is also negligible among Indian farmers due to their affordability to access the same. Very little research has been conducted to establish an exposure assessment procedure through dermal penetration of pesticide residues. Therefore, to quantify the contamination of pesticide residues through dermal exposure along with detailed field observations and pesticide management practices, a field study was conducted in Rangareddy district, Telangana, Southern India, to assess the dermal exposure based on dosimeter and hand washing methods. The analytical method was modified and validated in-house for performance parameters such as limit of detection, quantification, linear range, recovery, and precision. The potential dermal exposure values ranged from 0.15 to 13.45 µg, while a reduction was found in exposure levels as actual dermal exposure values ranged from 0 to 0.629 µg. Contamination through hand washing was the major contributor to overall dermal exposure. Statistical analysis revealed a significant difference in the exposed dermal regions of the leg and torso after the use of PPE. Penetration factor for each anatomical region and risk evaluation in terms of the Margin of Safety implies unsafe handling of pesticides. The findings of the present study confirm the increased exposure to organophosphate pesticides among operators and highlight the importance of the use of protective measures, especially among those that focus on dermal exposure mitigation.

Impact of Micronutrient Supplementation on Pesticide Residual, Acetylcholinesterase Activity, and Oxidative Stress Among Farm Children Exposed to Pesticides.

The present interventional study aimed to assess the impact of micronutrient supplementation on pesticide-residues concentrations, vitamins, minerals, acetylcholinesterase activity and oxidative stress among 129 farm children (9-12 years, n = 66 and 13-15 years, n = 63) involved in farming activities in Ranga Reddy district, Telangana, India. Our data showed the presence of five organophosphorus pesticide residues (chlorpyrifos, diazinon, malathion, monocrotophos, and phosalone) among children before-supplementation (both age-groups); while post-supplementation, only two pesticide residues (chlorpyrifos and diazinon) were detected indicating improved metabolic rate. Vitamin E, copper, magnesium and zinc levels were also improved in both the age-groups and manganese levels were significantly increased only among children of 13-15 years age group. Further, post-supplementation also showed an improvement in acetylcholinesterase activity and a decrease in lipid peroxidation among both the age groups of children. However, further research for ascertaining the ameliorating effect of micronutrients in preventing adverse effects of organophosphorus pesticides must be conducted.

Alterations in reproductive hormone levels among farm women and their children occupationally exposed to organophosphate pesticides.

Exposure to organophosphorus pesticides may lead to reproductive hormone dysfunction. Even among children of pubertal age, the exposure may disrupt growth, development, and maturation. The present study was conducted to assess the alterations in the reproductive hormone levels, among farm women (24-45 years, n = 129) and their children (9-12 years, n = 66 and 13-15 years, n = 63) and compare them with age and gender-matched control group [women (n = 134) and their children (9-12 years, n = 69 and 13-15 years, n = 65)] belonging to villages of Ranga Reddy District, Telangana, India. Blood pesticide residues and reproductive hormone (follicle-stimulating hormone-FSH, luteinizing hormone-LH, estradiol, and testosterone) levels were analyzed. The detected pesticide residues (ng/mL) were chlorpyrifos, diazinon, malathion, and monocrotophos among the farm women, while the farm children of 9-12 years age groups were detected with residues of chlorpyrifos, diazinon, malathion, monocrotophos, and phosalone. The farm children of 13-15 years age group were detected with residues of chlorpyrifos, diazinon, malathion, monocrotophos, and phosalone. However, no residues were detected among the samples of women and children of control groups. Significantly lower levels of FSH (in follicular phase) were observed among the farm women than the control group. Significant alterations in FSH and LH levels of farm women were observed with a significant correlation between the chlorpyrifos residue levels and estradiol hormone. While no such significant change in hormone levels was observed among the farm children of both age groups of both genders. Though the present study showed pesticide-induced alterations in hormone levels among the farm women, research is needed to elucidate the critical windows during which exposure may adversely affect the reproductive system in children at the pubertal stage and women at reproductive age and subsequently their progeny's health at a later stage of life.

Is hepatocellular carcinoma complicated with portal vein tumor thrombosis potentially curable by radiotherapy in the form of stereotactic body radiation therapy?

PURPOSE: The prognosis of hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) is dismal. Despite best treatment and care, the patients with this malignancy only showed 2.7-4 months of overall survival. It is debatable whether liver transplantation helps PVTT sufferers. The effectiveness of radiation therapy in treating HCC patients with PVTT should not be undervalued. By limiting the high dosage region to a small planning target volume, stereotactic radiation delivery has shifted toward hypofractionation, limiting the radiation exposure to healthy organs and tissues. Stereotactic body radiotherapy (SBRT) has a local control rate of 75-100%, depending on the treatment. The major limitation in SBRT for hepatocellular carcinoma with PVTT is the paucity of prospective evidence for longer periods beyond the first two years after treatment. More prospective studies/randomized clinical trials with a longer follow-up, larger sample size, and adequate statistical power are the dire need of the present situation to ascertain the curative effect of SBRT as primary therapy for advanced HCC with PVTT. CONCLUSION: SBRT can improve survival, particularly for patients receiving multidisciplinary treatment. This review sums up our most current understanding of how radiation therapy, notably SBRT, can be used to treat hepatocellular carcinoma when combined with PVTT. Recent research has led us to believe that irradiation in the form of SBRT may cure hepatocellular carcinoma complicated by PVTT.

Computational study of novel inhibitory molecule, 1-(4-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)piperazin-1-yl)-3-phenylurea, with high potential to competitively block ATP binding to the RNA dependent RNA polymerase of SARS-CoV-2 virus.

For coronaviruses, RNA-dependent RNA polymerase (RdRp) is an essential enzyme that catalyses the replication from RNA template and therefore remains an attractive therapeutic target for anti-COVID drug discovery. In the present study, we performed a comprehensive in silico screening for 16,776 potential molecules from recently established drug libraries based on two important pharmacophores (3-amino-4-phenylbutan-2-ol and piperazine). Based on initial assessment, 4042 molecules were obtained suitable as drug candidates, which were following Lipinski's rule. Molecular docking implemented for the analysis of molecular interactions narrowed this number of compounds down to 19. Subsequent to screening filtering criteria and considering the critical parameters viz. docking score and MM-GBSA binding free energy, 1-(4-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)piperazin-1-yl)-3-phenylurea (compound 1) was accomplished to score highest in comparison to the remaining 18 shortlisted drug candidates. Notably, compound 1 displayed higher docking score (-8.069 kcal/mol) and MM-GBSA binding free energy (-49.56 kcal/mol) than the control drug, remdesivir triphosphate, the active form of remdesivir as well as adenosine triphosphate. Furthermore, a molecular dynamics simulation was carried out (100 ns), which substantiated the candidacy of compound 1 as better inhibitor. Overall, our systematic in silico study predicts the potential of compound 1 to exhibit a more favourable specific activity than remdesivir triphosphate. Hence, we suggest compound 1 as a novel potential drug candidate, which should be considered for further exploration and validation of its potential against SARS-CoV-2 in wet lab experimental studies.Communicated by Ramasawamy H. Sarma.

Heat Shock Proteins and Their Role in Pregnancy: Redefining the Function of "Old Rum in a New Bottle".

Pregnancy in humans is a multi-step complex physiological process comprising three discrete events, decidualization, implantation and placentation. Its overall success depends on the incremental advantage that each of the preceding stages passes on to the next. The success of these synchronized sequels of events is an outcome of timely coordination between them. The pregnancy events are coordinated and governed primarily by the ovarian steroid hormones, estrogen and progesterone, which are essentially ligand-activated transcription factors. It's well known that intercellular signaling of steroid hormones engages a plethora of adapter proteins that participate in executing the biological functions. This involves binding of the hormone receptor complex to the DNA response elements in a sequence specific manner. Working with Drosophila melanogaster, the heat shock proteins (HSPs) were originally described by Ferruccio Ritossa back in the early 1960s. Over the years, there has been considerable advancement of our understanding of these conserved families of proteins, particularly in pregnancy. Accumulating evidence suggests that endometrial and uterine cells have an abundance of HSP27, HSP60, HSP70 and HSP90, implying their possible involvement during the pregnancy process. HSPs have been found to be associated with decidualization, implantation and placentation, with their dysregulation associated with implantation failure, pregnancy loss and other feto-maternal complications. Furthermore, HSP is also associated with stress response, specifically in modulating the ER stress, a critical determinant for reproductive success. Recent advances suggest a therapeutic role of HSPs proteins in improving the pregnancy outcome. In this review, we summarized our latest understanding of the role of different members of the HSP families during pregnancy and associated complications based on experimental and clinical evidences, thereby redefining and exploring their novel function with new perspective, beyond their prototype role as molecular chaperones.

Immunology of HPV-mediated cervical cancer: current understanding.

Human papilloma virus (HPV) has emerged as a primary cause of cervical cancer worldwide. HPV is a relatively small (55 nm in diameter) and non-enveloped virus containing approximately 8 kb long double stranded circular DNA genome. To date, 228 genotypes of HPV have been identified. Although all HPV infections do not lead to the development of malignancy of cervix, only persistent infection of high-risk types of HPV (mainly with HPV16 and HPV18) results in the disease. In addition, the immunity of the patients also acts as a key determinant in the carcinogenesis. Since, no HPV type specific medication is available for the patient suffering with cervical cancer, hence, a deep understanding of the disease etiology may be vital for developing an effective strategy for its prevention and management. From the immunological perspectives, the entire mechanisms of disease progression still remain unclear despite continuous efforts. In the present review, the recent developments in immunology of HPV-mediated cervix carcinoma were discussed. At the end, the prevention of disease using HPV type specific recombinant vaccines was also highlighted.

Predominant role of antioxidants in ameliorating the oxidative stress induced by pesticides.

Oxidative stress has been reported as one of the adverse effects caused due to pesticides, which is the main mechanism of the toxicity in humans and animals and is a useful parameter in monitoring studies. It involves an imbalance in the equilibrium state of ROS and antioxidant defenses leading to alterations in various antioxidant enzyme levels and lipid peroxidation. The objective of the current paper is to present a review of the potential role and protective mechanism action of the antioxidant micronutrient supplementation to ameliorate the oxidative stress induced by pesticides. Studies in animal models and human were retrieved through the relevant search of the literature and categorized. Various animal studies were categorized according to the type of supplementation. Animal studies provide evidence to conclude the potential protective role of antioxidants in ameliorating the adverse effects of pesticides. Similar studies in humans are meager suggesting for further comprehensive research.

Understanding the early host immune response against Mycobacterium tuberculosis.

Generation of immune response is a crucial activity of host defense against any microbial attack. When facultative organism Mycobacterium tuberculosis (MTB) invades its host, various pathways are activated in the host to mount immune responses against invading pathogen for nullifying its actions. During this host-pathogen interaction, interplay of complex network of cytokines and chemokines, initiation of phagocytosis, and formation of granuloma play an important role in containing MTB infections at host side. Simultaneously, MTB also evolves a plethora of specialized mechanisms to evade the host's killing cascades on other side, and during this bilateral cross-talk, many mycobacterial products play crucial role in survival of MTB inside the host. Hence, a better understanding of these phenomena is necessary not only for getting clear picture of pathogenesis of MTB, but also for developing effective, preventive, and therapeutic modalities against the pathogen. With some suggestions on future work, an insight into diversity of immune response of host against MTB was provided in the present review.

How homeopathic medicine works in cancer treatment: deep insight from clinical to experimental studies.

OBJECTIVE: In the current scenario of medical sciences, homeopathy, the most popular system of therapy, is recognized as one of the components of complementary and alternative medicine (CAM) across the world. Despite, a long debate is continuing whether homeopathy is just a placebo or more than it, homeopathy has been considered to be safe and cost-effectiveness therapeutic modality. A number of human ailments ranging from common to serious have been treated with homeopathy. However, selection of appropriate medicines against a disease is cumbersome task as total spectrum of symptoms of a patient guides this process. Available data suggest that homeopathy has potency not only to treat various types of cancers but also to reduce the side effects caused by standard therapeutic modalities like chemotherapy, radiotherapy or surgery. Although homeopathy has been widely used for management of cancers, its efficacy is still under question. In the present review, the anti-cancer effect of various homeopathic drugs against different kinds of cancers has been discussed and future course of action has also been suggested.

IL-10 down-regulates the expression of survival associated gene hspX of Mycobacterium tuberculosis in murine macrophage

Abstract Mycobacterium tuberculosis (MTB) adopts a special survival strategy to overcome the killing mechanism(s) of host immune system. Amongst the many known factors, small heat shock protein 16.3 (sHSP16.3) of MTB encoded by gene hspX has been reported to be critical for the survival of MTB. In the present study, the effect of recombinant murine interferon-gamma (rmIFN-γ) and recombinant murine interleukin-10 (rmIL-10) on the expression of gene hspX of MTB in murine macrophage RAW264.7 has been investigated. By real-time RT-PCR, it was observed that three increasing concentrations (5, 25 and 50 ng/ml) of rmIFN-γ significantly up-regulated the expression of hspX whereas similar concentrations of rmIL-10 (5, 25 and 50 ng/ml) significantly down-regulated the hspX expression. This effect was not only dependent on the concentration of the stimulus but this was time-dependent as well. A contrasting pattern of hspX expression was observed against combinations of two different concentrations of rmIFN-γ and rmIL-10. The study results suggest that rIL-10 mediated down-regulation of hspX expression, in the presence of low concentration of rIFN-γ, could be used as an important strategy to decrease the dormancy of MTB in its host and thus making MTB susceptible to the standard anti-mycobacterial therapy used for treating tuberculosis. However, as these are only preliminary results in the murine cell line model, this hypothesis needs to be first validated in human cell lines and subsequently in animal models mimicking the latent infection using clinical isolates of MTB before considering the development of modified regimens for humans.

IL-10 down-regulates the expression of survival associated gene hspX of Mycobacterium tuberculosis in murine macrophage.

Mycobacterium tuberculosis (MTB) adopts a special survival strategy to overcome the killing mechanism(s) of host immune system. Amongst the many known factors, small heat shock protein 16.3 (sHSP16.3) of MTB encoded by gene hspX has been reported to be critical for the survival of MTB. In the present study, the effect of recombinant murine interferon-gamma (rmIFN-γ) and recombinant murine interleukin-10 (rmIL-10) on the expression of gene hspX of MTB in murine macrophage RAW264.7 has been investigated. By real-time RT-PCR, it was observed that three increasing concentrations (5, 25 and 50ng/ml) of rmIFN-γ significantly up-regulated the expression of hspX whereas similar concentrations of rmIL-10 (5, 25 and 50ng/ml) significantly down-regulated the hspX expression. This effect was not only dependent on the concentration of the stimulus but this was time-dependent as well. A contrasting pattern of hspX expression was observed against combinations of two different concentrations of rmIFN-γ and rmIL-10. The study results suggest that rIL-10 mediated down-regulation of hspX expression, in the presence of low concentration of rIFN-γ, could be used as an important strategy to decrease the dormancy of MTB in its host and thus making MTB susceptible to the standard anti-mycobacterial therapy used for treating tuberculosis. However, as these are only preliminary results in the murine cell line model, this hypothesis needs to be first validated in human cell lines and subsequently in animal models mimicking the latent infection using clinical isolates of MTB before considering the development of modified regimens for humans.