Manas Ranjan Ray

Systemic inflammatory changes and increased oxidative stress in rural Indian women cooking with biomass fuels.

The study was undertaken to investigate whether regular cooking with biomass aggravates systemic inflammation and oxidative stress that might result in increase in the risk of developing cardiovascular disease (CVD) in rural Indian women compared to cooking with a cleaner fuel like liquefied petroleum gas (LPG). A total of 635 women (median age 36 years) who cooked with biomass and 452 age-matched control women who cooked with LPG were enrolled. Serum interleukin-6 (IL-6), C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8) were measured by ELISA. Generation of reactive oxygen species (ROS) by leukocytes was measured by flow cytometry, and erythrocytic superoxide dismutase (SOD) was measured by spectrophotometry. Hypertension was diagnosed following the Seventh Report of the Joint Committee. Tachycardia was determined as pulse rate >100 beats per minute. Particulate matter of diameter less than 10 and 2.5 μm (PM₁₀ and PM₂.₅, respectively) in cooking areas was measured using real-time aerosol monitor. Compared with control, biomass users had more particulate pollution in indoor air, their serum contained significantly elevated levels of IL-6, IL-8, TNF-α and CRP, and ROS generation was increased by 37% while SOD was depleted by 41.5%, greater prevalence of hypertension and tachycardia compared to their LPG-using neighbors. PM₁₀ and PM₂.₅ levels were positively associated with markers of inflammation, oxidative stress and hypertension. Inflammatory markers correlated with raised blood pressure. Cooking with biomass exacerbates systemic inflammation, oxidative stress, hypertension and tachycardia in poor women cooking with biomass fuel and hence, predisposes them to increased risk of CVD development compared to the controls. Systemic inflammation and oxidative stress may be the mechanistic factors involved in the development of CVD.

Hypertension with elevated levels of oxidized low-density lipoprotein and anticardiolipin antibody in the circulation of premenopausal Indian women chronically exposed to biomass smoke during cooking.

UNLABELLED This study aims to investigate whether indoor air pollution (IAP) from biomass fuel use was associated with hypertension, platelet hyperactivity, and elevated levels of oxidized low-density lipoprotein (oxLDL) and anticardiolipin antibody (aCL). We enrolled 244 biomass fuel-using (median age 34 year) and 236 age-matched control women who cooked with liquefied petroleum gas (LPG). Enzyme-linked immunosorbent assay was used to measure oxLDL in plasma and aCL in serum, flow cytometry for P-selectin expression on platelet and reactive oxygen species (ROS) generation by leukocytes, aggregometry for platelet aggregation, spectrophotometry for superoxide dismutase (SOD) in erythrocytes, and laser photometer for particulate matter <10 and 2.5 μm in diameter (PM(10) and PM(2.5), respectively) in cooking areas. Biomass users had three times more particulate pollution in kitchen, had higher prevalence of hypertension (29.5 vs. 11.0% in control, P < 0.05), elevated oxLDL (170.6 vs. 45.9 U/l; P < 0.001), platelet P-selectin expression (9.1% vs. 2.4%), platelet aggregation (23.2 vs. 15.9 Ohm), raised aCL IgG (28.7% vs. 2.1%), IgM (8.6% of vs. 0.4%), and ROS (44%) but depleted (13%) SOD. After controlling potential confounders, the changes were positively associated with PM(10) and PM(2.5) in indoor air, suggesting a positive association between IAP and increased cardiovascular risk. PRACTICAL IMPLICATIONS The study showing high risk of developing cardiovascular diseases (CVD) among poor, underprivileged women in their reproductive ages in rural India is important from public health perspectives. It may motivate the government and the regulatory agencies of the country to take a serious note of the indoor air pollution (IAP) from biomass fuel use as it threatens the health of millions of women, children, and the elderly who mostly stay indoor. We hope the findings will strengthen the demand for setting up a standard for indoor air quality in the country in the line of national ambient air quality standard. The findings may also inspire the authorities to take measures for the reduction in IAP by improving housing, kitchen ventilation, and cook stoves. Moreover, the parameters used in this study can be utilized for large, population-based studies to identify women at a higher risk of developing CVD so that medical intervention can be taken at the formative stage of a disease.

Platelet activation, upregulation of CD11b/ CD18 expression on leukocytes and increase in circulating leukocyte-platelet aggregates in Indian women chronically exposed to biomass smoke

The majority of households in rural India still rely on unprocessed solid biomass for domestic energy. The aim of this study was to investigate whether chronic exposure to biomass smoke causes activation of leukocytes and the formation of leukocyte-platelet aggregates. We conducted flow cytometric analysis of β2 Mac-1 integrin (CD11b/CD18) expression on polymorphonuclear leukocytes (PMN) and monocytes, and P-selectin (CD62P) expression on the platelets of 165 women from eastern India, who cook solely with wood, dung and agricultural wastes, and 155 age- and socio-economic condition-matched control subjects, who used relatively cleaner fuel, liquefied petroleum gas (LPG). Leukocyte-platelet aggregates were defined as CD11b-positive PMN and monocytes co-expressing platelet-specific markers CD41 or CD62P. A significant increase in leukocyte-platelet aggregates was found in women who used biomass as cooking fuel. In addition, they showed increased surface expression of CD11b/CD18 in circulating PMN and monocytes and CD62P expression on platelets. The mean fluorescence intensity (MFI) of CD11b on the surface of circulating monocytes and PMN of biomass users increased by 50 and 68%, respectively. Similarly, a 62 and 48% increase in MFI was observed in CD18 expression on the surface of these cells in biomass users. The results show that chronic biomass smoke exposure activates circulating platelets, PMN and monocytes, and increases the number of leukocyte-platelet aggregates, which are considered a risk factor for thrombosis.

Chronic Exposures to Cholinesterase-inhibiting Pesticides Adversely Affect Respiratory Health of Agricultural Workers in India

Chronic Exposures to Cholinesterase‐inhibiting Pesticides Adversely Affect Respiratory Health of Agricultural Workers in India: Sreeparna Chakraborty, et al. Department of Experimental Hematology, Chittaranjan National Cancer Institute, India

Micronucleus formation, DNA damage and repair in premenopausal women chronically exposed to high level of indoor air pollution from biomass fuel use in rural India

Genotoxicity of indoor air pollution from biomass fuel use has been examined in 132 biomass users (median age 34 years) and 85 age-matched control women from eastern India who used the cleaner fuel liquefied petroleum gas (LPG) to cook. Micronucleus (MN) frequency was evaluated in buccal (BEC) and airway epithelial cells (AEC); DNA damage was examined by comet assay in peripheral blood lymphocytes (PBL); and expressions of gamma-H2AX, Mre11 and Ku70 proteins were localized in AEC and PBL by immunocytochemistry. Reactive oxygen species (ROS) generation in leukocytes was measured by flow cytometry, and the levels of superoxide dismutase (SOD) and total antioxidant status (TAS) in blood were measured by spectrophotometry. Real-time aerosol monitor was used to measure particulate pollutants in indoor air. Compared with controls, biomass users had increased frequencies of micronucleated cells in BEC (3.5 vs. 1.7, p<0.001) and AEC (4.54 vs. 1.86, p<0.001), and greater comet tail % DNA (18.6 vs. 11.7%, p<0.01), tail length (45.5 vs. 31.4mum, p<0.01) and olive tail moment (4.0 vs. 1.4, p<0.01) in PBL. Moreover, biomass users had more gamma-H2AX-positive nuclei in PBL (49.5 vs. 8.5%, p<0.01) and AEC (11.3 vs. 2.9%, p<0.01) along with higher expression of DNA repair proteins Mre11 and Ku70 in these cells, suggesting stimulation of DNA repair mechanism. Biomass users showed rise in ROS generation and depletion of SOD and TAS. Biomass-using households had 2-4 times more particulate matter with diameter less than 10 and 2.5mum in indoor air, and MN frequency and comet tail % DNA were positively associated with these pollutants after controlling potential confounders. Thus, chronic exposure to biomass smoke causes chromosomal and DNA damage and upregulation of DNA repair mechanism.

Assessment of DNA damage by comet assay and fast halo assay in buccal epithelial cells of Indian women chronically exposed to biomass smoke

Genotoxicity of indoor air pollution from biomass burning was evaluated in buccal epithelial cells (BECs) of 85 pre-menopausal Indian women who were engaged in cooking with biomass (wood, dung, crop residues) and 76 age-matched control women who were cooking with cleaner fuel liquefied petroleum gas (LPG). DNA damage was evaluated by comet assay and fast halo assay (FHA). The concentrations of particulate matter with aerodynamic diameters of less than 10 and 2.5 μm (PM(10) and PM(2.5), respectively) in indoor air were measured by real-time aerosol monitor. Generation of reactive oxygen species (ROS) was measured by flow cytometry and the level of superoxide dismutase (SOD) by spectrophotometry. Compared with control, BEC of biomass users illustrated 2.6-times higher comet tail % DNA (32.2 vs. 12.4, p < 0.001), 2.7-times greater comet tail length (37.8 μm vs. 14.2 μm, p < 0.001) and 2.2-times more olive tail moment (7.1 vs. 3.2, p < 0.001), suggesting marked increase in DNA damage. FHA also showed 5-times more mean nuclear diffusion factor (9.2 vs. 1.8, p < 0.0001) in BEC of biomass users, confirming sharp rise in DNA single strand breaks. Airway cells of biomass-using women showed 51% rise in ROS generation but 28% reduction in SOD, suggesting oxidative stress in the airways. Indoor air of biomass-using households had 3-times more PM(10) and PM(2.5) than LPG-using families, and DNA damage showed positive association with PM(10) and PM(2.5) levels controlling education, kitchen location and family income as potential confounders. In summary, chronic inhalation of biomass smoke elicits oxidative stress and extensive DNA damage in BEC.

Immune cells and cardiovascular health in premenopausal women of rural India chronically exposed to biomass smoke during daily household cooking

Changes in cells of the immune system are important indicators of systemic response of the body to air pollution. The aim of this study was to investigate the immunological changes in rural women who have been cooking exclusively with biomass for the past 5 years or more and compare the findings with women cooking exclusively with liquefied petroleum gas (LPG). We conducted a cross-sectional analysis of the associations between indices of indoor air pollution (IAP) and a set of immune assays. Biomass users illustrated marked suppression in the total number of T-helper (CD4+) cells and B (CD19+) cells while appreciable rise was documented in the number of CD8+ T-cytotoxic cells and CD16+CD56+ natural killer (NK) cells. A consistent finding among biomass users was rise in regulatory T (Treg) cells. Among biomass users, peripheral lymphocyte subpopulations, Treg cells, and the number of typical monocytes (CD16-CD64+ cells), antigen presenting types (CD16+CD64- cells) and plasmacytoid cells (CD16-CD64- cells) were found to be significantly altered in those who daily cooked with dung in comparison to wood and crop residue users (p<0.05). Biomass users who cooked in kitchens adjacent to their living areas had significant changes in peripheral lymphocyte subpopulations, typical monocytes (CD16-CD64+) with high phagocytic activity and antigen presenting monocytes (CD16+CD64-) against women who cooked in separate kitchens (p<0.01). This study has shown that women who cooked exclusively with biomass fuel had alterations in immune defense compared with their neighbors who cooked with LPG.

Changes in sputum cytology, airway inflammation and oxidative stress due to chronic inhalation of biomass smoke during cooking in premenopausal rural Indian women

To perform sputum analysis for verification of pulmonary changes in premenopausal rural Indian women chronically exposed to biomass smoke during cooking.Three consecutive morning sputum samples were collected from 196 women (median age 34 years) cooking with biomass and 149 age-matched control women cooking with cleaner fuel liquefied petroleum gas. Smears made on slides were stained with Papanicolaou and Perls Prussian blue. Airway oxidative stress was estimated as reactive oxygen species (ROS) generation (by flow cytometry) and superoxide dismutase (SOD) level (by spectrophotometry) in sputum cells. Airway inflammation was measured as sputum levels of interleukin (IL)-6, -8 and tumor necrosis factor- alpha (TNF-α). Particulate matter of diameter less than 10 (PM10) was measured using laser photometer while benzene exposure was monitored by measuring trans, trans-muconic acid (t,t-MA) in urine by HPLC-UV. Compared with control, sputum of biomass users contained more neutrophils, lymphocytes, eosinophils, alveolar macrophages, and showed presence of ciliocytophthoria, Charcot-Leyden crystals, Curschmanns spiral. ROS generation was increased by 2-fold while SOD was depleted by 31% in biomass users. They also had higher sputum levels of IL-6, -8 and TNF-α. Levels of PM10 and t,t-MA were 2.9- and 5.8-times higher in biomass-using women. PM10 and t,t-MA levels were positively associated with cellular changes in the sputum, markers of airway inflammation, and oxidative stress. Cooking with biomass alters sputum cytology, and increases airway inflammation and oxidative stress that might result in further amplification of the tissue damaging cascade in women chronically exposed to biomass smoke.

Chronic inhalation of biomass smoke is associated with DNA damage in airway cells: involvement of particulate pollutants and benzene.

This study examined whether indoor air pollution from biomass fuel burning induces DNA damage in airway cells. For this, sputum cells were collected from 56 premenopausal rural women who cooked with biomass (wood, dung, crop residues) and 49 age‐matched controls who cooked with cleaner liquefied petroleum gas. The levels of particulate matters with diameters of less than 10 and 2.5 µm (PM10 and PM2.5) in indoor air were measured using a real‐time aerosol monitor. Benzene exposure was monitored by measuring trans,trans‐muconic acid (t,t‐MA) in urine by HPLC‐UV. DNA damage was examined by alkaline comet assay in sputum cells. Generation of reactive oxygen species (ROS) and level of superoxide dismutase (SOD) in sputum cells were measured by flow cytometry and spectrophotometry, respectively. Compared with controls, biomass users had 4 times higher tail percentage DNA, 37% more comet tail length and 5 times more Olive tail moment (p < 0.001) in inflammatory and epithelial cells in sputum, suggesting extensive DNA damage. In addition, women who cooked with biomass had 6 times higher levels of urinary t,t‐MA and 2‐fold higher levels of ROS generation concomitant with 28% depletion of SOD. Indoor air of biomass‐using households had 2–4 times more PM10 and PM2.5 than that of controls. After controlling potential confounders, positive association was found between DNA damage parameters, particulate pollution, urinary t,t‐MA and ROS. Thus, long‐term exposure to biomass smoke induces DNA damage in airway cells and the effect was probably mediated, at least in part, by oxidative stress generated by inhaled particulate matter and benzene. Copyright

Prevalence of hypertension and pre-hypertension in rural women: a report from the villages of West Bengal, a state in the eastern part of India.

OBJECTIVE To find out the prevalence of hypertension, pre-hypertension and tachycardia among the women in rural areas of West Bengal, identify co-factors associated with the prevalence and contribute to the body of evidence for future health programs to identify at-risk groups. DESIGN A population-based cross-sectional study was conducted. SETTING The study was conducted in remote villages. PARTICIPANTS 1186 women participants, aged 18 years or more were included. MAIN OUTCOME MEASURES They were interviewed using standard structured questionnaire. Blood pressure and tachycardia was monitored using digital sphygmomanometer. For each participant, we made two blood pressure measurements with an interval of 48 hours. Data was analysed statistically using SPSS software. RESULTS Overall prevalence of hypertension in the study subjects was 24.7% and that of pre-hypertension and tachycardia was 40.8% and 6.4%, respectively. Both hypertension and pre-hypertension were seen to increase with age. Other identified significant factors were use of biomass fuel for cooking, absence of separate kitchen, higher body mass index (BMI), education and average family income. CONCLUSION This study suggests quite high prevalence of hypertension as well as pre-hypertension among the women of rural areas. The findings are significant from the women health perspectives. Early detection of pre-hypertensive and hypertensive subjects will help to formulate intervention strategies to allay the spread of cardiovascular diseases.