Production and recovery of polyhydroxyalkanoates (PHA) from waste streams - A review.

Polyhydroxyalkanoates (PHA) are the more attractive sustainable green plastic, and it has the potential to replace petroleum-based plastics (PBP) in the global market. Recently, most of the developed and developing countries have banned the use of traditional PBP. This increases the demand for green plastic production and positively impacts the global market. Producing green plastic from various waste streams such as whey, animal, and crude glycerol will be eco-friendly and cost-effective. However, the factors influencing the environmental sustainability of PHA production from different waste streams are still unclear. This review could be reinforced concrete to researchers to gather deep knowledge on techno-economic analysis, life-cycle assessment, environmental and ecological risks caused during PHA production from different waste streams.

Evaluation of global research trends in photocatalytic degradation of dye effluents using scientometrics analysis.

Photocatalysis technology is observed to be an effective approach for its outstanding performance to eliminate wide range of organic pollutants including dyes in textile effluent. Despite growing number of studies, there is no scientometric perspective addressing the research topic "photocatalytic degradation of dye effluents". In this regard, a total of 954 documents were extracted from the Web of Science (WoS) database using keywords search to cover all the published documents during the period 1996-2020. Publications in this area started to increase exponentially from year 2007. The most dominant subject categories were Engineering, Chemistry and Environmental Science & Ecology. Applied Catalysis B-Environmental and Desalination & Water Treatment were identified as the most-impactful and productive journals respectively. Authors based in India accounted for 29.6% of total publications followed by China (14.2%); but in terms of citations Spain and Italy were more influential. Based on keyword analysis, azo dyes, TiO2, nanoparticles, adsorption, methylene blue, visible light, ZnO and kinetics are the most studied, and visible light mediated photocatalysis, hybrid treatment systems, nano based photocatalysis and more recently, metal based photocatalysis, have received most attention. Studies on cost and energy analysis, recovery of value-added products, development of more efficient photocatalytic materials and new photocatalyst regeneration approaches should be considered for future research. This study therefore, provides a comprehensive understanding about the trends and patterns of the specified research field worldwide.

Toxicoproteomic analysis of human lung epithelial cells exposed to steel industry ambient particulate matter (PM) reveals possible mechanism of PM related carcinogenesis.

Toxicoproteomic analysis of steel industry ambient particulate matter (PM) that contain high concentrations of PAHs and metals was done by treating human lung cancer cell-line, A549 and the cell lysates were analysed using quantitative label-free nano LC-MS/MS. A total of 18,562 peptides representing 1576 proteins were identified and quantified, with 196 proteins had significantly altered expression in the treated cells. Enrichment analyses revealed that proteins associated to redox homeostsis, metabolism, and cellular energy generation were inhibited while, proteins related to DNA damage and repair and other stresses were over expressed. Altered activities of several tumor associated proteins were observed. Protein-protein interaction network and biological pathway analysis of these differentially expressed proteins were carried out to obtain a systems level view of proteome changes. Together it could be inferred that PM exposure induced oxidative stress which could have lead into DNA damage and tumor related changes. However, lowering of cellular metabolism, and energy production could reduce its ability to overcome these stress. This kind of disequilibrium between the DNA damage and ability of the cells to repair the DNA damage may lead into genomic instability that is capable of acting as the driving force during PM induced carcinogenesis.

Fe0 catalyzed photo-Fenton process to detoxify the biodegraded products of azo dye Mordant Yellow 10.

Inspired by the efficiency of the photo-Fenton process on oxidation of organic pollutants, we herein present the feasibility of visible light driven photo-Fenton process as a post treatment of biological method for the effective degradation and detoxification of monoazo dye Mordant Yellow 10 (MY10). Anaerobic degradation of MY10 by Pseudomonas aeroginosa formed aromatic amines which were further degraded in the subsequent Fe catalyzed photo-Fenton process carried out at pH 3.0, with iron shavings and H2O2 under blue LED light illumination. LC-MS and stoichiometric analysis confirmed that reductive azo bond cleavage was the major reaction in anaerobic bacterial degradation of MY10 producing 4-amino benzene sulfonic acid (4-ABS) and 5-amino salicylic acid (5-ASA) which were further degraded into hydroxyl amines, nitroso and di/tri carboxylic acids by the photo-Fenton process. Toxicity studies with human small cell lung cancer A549 cells provide evidence that incorporation of Fe0 catalyzed photo-Fenton step after anaerobic bacterial treatment improved the mineralization and detoxification of MY10 dye.

Effect of size and shape on toxicity of zinc oxide (ZnO) nanomaterials in human peripheral blood lymphocytes.

The multi-industrial applications of zinc oxide nanomaterials (ZnO NMs) lead to increasing exposure to humans. Though the ZnO nanoparticles (NPs) toxicity had been evaluated previously, toxicity of other forms of ZnO nanomaterials has not been evaluated. In this study, cytotoxicity and genotoxicity of four different types of ZnO NMs were evaluated using human peripheral blood lymphocytes (HPBL). In addition, the effect of anti-oxidants on ZnO NMs induced toxicity was also evaluated. Our results suggest that, size and shape of the nanomaterials have profound effects on their toxicity. The NPs and nanorods (NRs) possessed higher level of oxidative potential and ROS generation capacity than microparticles (MPs) and microrods (MRs). In contrast, MPs and MRs possessed higher level of lipid peroxidation capacity. The smaller NPs are more genotoxic while larger MPs and MRs were more cytotoxic in nature. Treatment with vitamin C or Quercetin significantly reduces the genotoxicity associated with ZnO NMs. The influence of size and shape in mediating NMs toxicity should be taken into account and the possible supplementation of anti-oxidants might mitigate the toxicity.

Solar catalysed activity against methyl orange dye, cytotoxicity activity of MCF-7 cell lines and identification of marker compound by HPTLC of Lagerstroemia speciosa.

The investigation was aimed to quantify the Gallic acid present in Lagerstroemia speciosa leaves (Lythraceae). The High-Performance Thin Layer Chromatography (HPTLC) quantification was performed for acetone (AE), methanolic (ME) and chloroform (CE) extract of leaves of L. speciosa. The pre-coated silica gel 60 F254 was used for complete separation of compounds using the mobile phase pet. Ether: ethyl acetate: formic acid (5:5:1v/v).The validation of the extracts was carried out using ICH guidelines for precision, repeatability and accuracy showing the Rf 0.49 against standard Gallic acid. Linearity range for Gallic acid was done from 200 to 1000ng/spot (AE) and200 ng to 600ng/spot (ME), with Correlation, coefficient r=0.99 (AE) and 0.54 (ME) in the said concentrations. The composition in crude leaf extract was determined to be of 49.712mg (AE) and 20.125mg (ME), while it was not found in chloroform extract against standard Gallic acid. Hence the proposed method was very simple, precise, accurate and easy for the screening of the bioactive compounds present in the acetone and methanolic extracts of the leaves of L. speciosa. It was observed that the acetone extract subjected to cytotoxicity showed promising activity at higher concentrations (100 and 200µg/ml) showed 92.9% and 87.13% inhibition against MCF-7 cell lines respectively. The photocatalytic activity of the acetone and methanolic extracts of methyl orange was found to be 90.25% (190min) and 89.03% (180min) respectively. Therefore this can be used as an indicator of purity of herbal drugs and formulation containing L. speciosa.

MicroRNA Regulation of Acute Lung Injury and Acute Respiratory Distress Syndrome.

The acute respiratory distress syndrome (ARDS), a severe form of acute lung injury (ALI), is a very common condition associated with critically ill patients, which causes substantial morbidity and mortality worldwide. Despite decades of research, effective therapeutic strategies for clinical ALI/ARDS are not available. In recent years, microRNAs (miRNAs), small non-coding molecules have emerged as a major area of biomedical research as they post-transcriptionally regulate gene expression in diverse biological and pathological processes, including ALI/ARDS. In this context, this present review summarizes a large body of evidence implicating miRNAs and their target molecules in ALI/ARDS originating largely from studies using animal and cell culture model systems of ALI/ARDS. We have also focused on the involvement of miRNAs in macrophage polarization, which play a critical role in regulating the pathogenesis of ALI/ARDS. Finally, the possible future directions that might lead to novel therapeutic strategies for the treatment of ALI/ARDS are also reviewed. J. Cell. Physiol. 231: 2097-2106, 2016. © 2016 Wiley Periodicals, Inc.

MicroRNAs as potential targets for progressive pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and devastating disorder. It is characterized by alveolar epithelial cell injury and activation, infiltration of inflammatory cells, initiation of epithelial mesenchymal transition (EMT), aberrant proliferation and activation of fibroblasts, exaggerated deposition of extracellular matrix (ECM) proteins, and finally leading to the destruction of lung parenchyma. MicroRNAs (miRNAs) are endogenous small non-coding RNA molecules that post-transcriptionally regulate gene expression in diverse biological and pathological processes, including cell proliferation, differentiation, apoptosis and metastasis. As a result, miRNAs have emerged as a major area of biomedical research with relevance to pulmonary fibrosis. In this context, the present review discusses specific patterns of dysregulated miRNAs in patients with IPF. Further, we discuss the current understanding of miRNAs involvement in regulating lung inflammation, TGF-ß1-mediated EMT and fibroblast differentiation processes, ECM genes expression, and in the progression of lung fibrosis. The possible future directions that might lead to novel therapeutic strategies for the treatment of pulmonary fibrosis are also reviewed.

Characterization and genotoxicity evaluation of particulate matter collected from industrial atmosphere in Tamil Nadu state, India.

Ambient particulate matter (PM) collected in the vicinity of five industries (Cement, Chemical, Thermal power plant, Sponge-iron and Steel) in Tamil Nadu state, India was characterized for size distribution, metals and polycyclic aromatic hydrocarbons (PAHs) content. Genotoxicity of PM and organic matter (OM) extracted from PM was measured in human lung cancer cell-line, A549 and in human liver carcinoma cell-line, HepG2, respectively, using the comet assay. PM values varied from 57.0 µg/m(3) of air at Cement industry upstream to 561.0 µg/m(3) of air at Sponge iron industry downstream samples. Their metal content varied from 5.758 µg/m(3) of air at Chemical industry to 46.144 µg/m(3) of air at Sponge iron industry and PAH concentration varied from 0.5 ng/m(3) air in upstream Thermal power plant to 3302.4 ng/m(3) air in downstream Sponge iron industry samples. While all PM samples induced DNA strand breaks at higher dose levels, downstream samples of Steel and Sponge iron industries which contained relatively higher concentrations of PAHs and metals and exhibited higher levels of pro-oxidant activity as measured by DTT activity induced significantly higher levels of DNA damage in HepG2 and A549 cells. Pretreatment of A549 cells with vitamin C or quercetin significantly reduced PM induced DNA strand breaks.

Protective effect of gallic acid on alloxan-induced oxidative stress and osmotic fragility in rats.

In the present study, we investigated the antioxidant effect of gallic acid (GA) on membrane lipid peroxidation and osmotic fragility in alloxan-induced diabetic Wistar rats. GA was administered orally at doses of 5, 10, and 20 mg/kg body weight for 45 days, after which liver and kidney tissues were analyzed for the degree of lipid peroxidation, reduced glutathione, and the activities of antioxidants such as catalase, superoxide dismutase, and glutathione peroxidase. Administration of GA to alloxan-induced diabetic rats reduced the blood glucose level with an increase in the level of insulin. Liver and kidney tissues from diabetic animals exhibited disturbances in antioxidant defense compared with normal rats. GA at a dose of 20 mg/kg b.w. showed a significant effect than that of the other doses. In addition, the results revealed that GA protected the integrity of erythrocyte membrane in diabetic rats as demonstrated by lower percentage of hemolysis and resistance to hydrogen peroxide-induced peroxidation. The anti-hyperglycemic activity of GA in alloxan-induced diabetic rats was also comparable with glibenclamide, a reference drug. These results suggest that GA could provide a beneficial effect on diabetes by decreasing oxidative stress-related diabetic complications.

In vitro cytotoxicity of Gymnema montanum in human leukaemia HL-60 cells; induction of apoptosis by mitochondrial membrane potential collapse.

OBJECTIVES: Gymnema montanum Hook, an Indian Ayurvedic medicinal plant, is used traditionally to treat a variety of ailments. Here, we report anti-cancer effects and molecular mechanisms of ethanolic extract of G. montanum (GLEt) on human leukaemia HL-60 cells, compared to peripheral blood mononuclear cells. MATERIALS AND METHODS: HL-60 cells were treated with different concentrations of GLEt (10-50 µg/ml) and cytotoxicity was assessed by MTT assay. Levels of lipid peroxidation, antioxidants, mitochondrial membrane potential and caspase-3 were measured. Further, apoptosis was studied using annexin-V staining and the cell cycle was analyzed by flow cytometry. RESULTS: GLEt had a potent cytotoxic effect on HL-60 cells (IC50 -20 µg/ml), yet was not toxic to normal peripheral blood mononuclear cells. Exposure of HL-60 cells to GLEt led to elevated levels of malonaldehyde formation, but to reduced glutathione, superoxide dismutase, catalase and glutathione peroxidase activities (P < 0.05). Induction of apoptosis was confirmed by observing annexin-V positive cells, associated with loss of mitochondrial membrane potential. Cell cycle arrest at G0/G1 was observed in GLEt-treated HL-60 cells, indicating its potential at inducing their apoptosis. CONCLUSIONS: Findings of the present study suggest that G. montanum induced apoptosis in the human leukaemic cancer cells, mediated by collapse of mitochondrial membrane potential, generation of reactive oxygen species and depletion of intracellular antioxidant potential.

The impact of oxidative stress on islet transplantation and monitoring the graft survival by non-invasive imaging.

Islet transplantation is an attractive strategy to treat severe diabetic conditions in patients suffering from autoimmune derived diabetes, and it has currently been considered a forefront research arena in diabetes. Major aim of islet transplantation is to achieve successful insulin independent disease free survival. The key challenges in transplanted islets are the generation of reactive oxygen species (ROS) and associated oxidative stress, pro-inflammatory cytokine - (TNFα) mediated apoptotic induction, attack by immune cells, and achieving revascularization with minimal hypoxic microenvironment. Free radicals and their derivatives are constantly produced in living systems, but at relatively low level, and in a balanced state. Oxidative stress, which occurs as a result of an imbalance between the intracellular free radicals production and the cellular antioxidant defense mechanisms in the transplanted islets, can lead to cell death. The balance between oxidants and antioxidants in a cell can be easily disturbed by increase in ROS production or reduction in the level of cellular antioxidant defensive substances, which can cause many metabolic complications, including pancreatic ß-cell damage. Antioxidants function as blockers of radical processes by eliminating harmful ROS produced during normal cellular metabolism. A complex antioxidant defense mechanism has been developed by nature in cells to protect the cellular homeostasis. This system mainly includes antioxidant enzymes, vitamins and minerals. As transplanted islet survival is crucial for achieving successful therapy, most of these antioxidants can be used as a supplement to scavenge the local ROS thereby improving the survival of transplanted islets. Currently, very few techniques have been routinely used to qualitatively and quantitatively assess the survival and function of islet grafts, especially to confirm the success of treatment, which includes metabolic parameters such as blood glucose, insulin and C-peptide levels. These biochemical measurements provide markers at only the late stages of islet rejection. Use of molecular imaging techniques has the potential for real-time non-invasive monitoring of the functional status and viability of transplanted islet grafts in living animals. This review mainly focuses on the current status of islet transplantations, potential preventive strategies used to reduce oxidative stress-mediated toxicity in islet grafts, and use of molecular imaging as a tool to quantitatively evaluate the functional status of the transplanted islets in living animals.

Antigenotoxic potential of Gymnema montanum leaves on DNA damage in human peripheral blood lymphocytes and HL-60 cell line.

In this study we have evaluated the genoprotective effect of the ethanol extract of Gymnema montanum (GLEt) leaves in human peripheral blood lymphocytes and HL-60 cell line in vitro using the comet assay. DNA damage was induced by treating the cells with H(2)O(2) and methyl methane sulphonate (MMS). GLEt treatment effectively protected the lymphocytes and HL-60 cell line from H(2)O(2)-induced oxidative DNA damage in a dose-dependent manner whereas it was not effective against alkylative DNA damage caused by MMS. The global percent repair efficiency also showed that both pre- and post- GLEt treatment provided effective protection against H(2)O(2) induced DNA damage but not as effective against MMS. At 200 microg ml(-1) level, its repair capacity against H(2)O(2) induced DNA damage was comparable to that of vitamin-C (100 microM). Furthermore, exposure to GLEt reduced the formation of apoptotic cells caused by H(2)O(2), which was demonstrated by the decreased sub-G1-DNA content in cell cycle analysis and apoptotic frequencies of lymphocytes in an annexin-V binding assay. In addition, GLEt was found to have effective peroxide scavenging ability in dose-dependent manner. The protective efficiency of the extract was found to be directly proportional to its total phenolic content. The present study indicates that G. montanum leaves are a significant source of phytochemicals with antigenotoxic and antioxidant activity, and thus has potential therapeutic use.

Assessment of heavy metal contents in the ambient air of the Coimbatore city, Tamilnadu, India.

Industrialization and urbanization are the two major causes of deteriorating air quality. To evaluate the ambient air quality of the Coimbatore city, suspended particulate matter (SPM) was collected at ten stations and analysed for the heavy metals content. The concentrations of seven heavy metals (Zn, Fe, Cu, Pb, Ni, Cr and Cd) were estimated. The level of SPM was found to be either at permissible or non-permissible limit depending upon the category of the sampling station. At majority of sampling stations, concentrations of Zn were found to be maximum than other heavy metals. The order of average concentrations of heavy metals in Coimbatore atmospheric air was Zn>Fe>Cu>Pb>Cr>Ni>Cd. The usage of Zn for protective coating on iron, steel etc. by the industries in Coimbatore city could be the major reason for the higher concentration of this heavy metal in this region.

Effect of Gymnema montanum leaves on red blood cell resistance to oxidative stress in experimental diabetes.

The aim of the present study was to evaluate the protective effect of Gymnema montanum on red blood cell (RBC) membrane in diabetic rats during lipid peroxidation. Ethanol extract of G. montanum leaves (GLEt) was administered orally to alloxan-induced diabetic rats for 3 weeks, and the effects on blood glucose, insulin, lipid peroxidation markers, thiobarbituric acid reactive substances, hydroperoxides in plasma and antioxidant enzymes including superoxide dismutase, catalase and glutathione peroxidase activities in erythrocytes were studied. Administration of GLEt to diabetic animals at doses of 50, 100, and 200 mg/kg body weight lowered elevated blood glucose levels by 24, 35, and 66%, respectively, relative to untreated diabetic rats. In comparison, treatment with the known antidiabetic drug, glibenclamide (600 microg/kg body weight) decreased blood glucose concentrations by 51%. Plasma insulin concentrations were increased in the diabetic rat by 73% with GLEt (200 mg/kg body weight) and 45% with glibenclamide (600 microg/kg body weight). Although a significant decrease in the lipid peroxidation markers was observed in plasma on treatment with GLEt and glibenclamide, the RBC antioxidant levels were increased significantly in diabetic rats. Furthermore, erythrocytes from the GLEt-treated animals were found to be more resistant to H2O2-induced peroxidation than that of untreated diabetic animals. The chemical characterization of the polyphenolics of the extract showed the presence of gallic acid (5.29% w/w), resveratrol (2.2% w/w), and quercetin (16.6% w/w). The results of this study suggest that G. montanum may be useful for the control, management, and prevention of oxidative stress associated with diabetes.

Genotoxicity of a polluted river system measured using the alkaline comet assay on fish and earthworm tissues.

Monitoring genotoxicity in the environment by using endemic organisms as sentinels requires sensitive assays. In this study the genotoxic properties of water and sediment collected from the Noyyal River, which is polluted with industrial effluent and sewage, was determined in fish (Cyprinus carpio) and earthworms (Eisenia foetida) using the alkaline comet assay. Upon electrophoresis, extensive DNA damage, measured as the DNA length:width ratio of the DNA mass, was observed in erythrocytes, liver, and kidney cells of fish exposed to polluted water samples and the amount of damage increased with the duration of exposure. Similarly, the mean DNA length:width ratio was significantly higher in the coelomocytes of earthworms placed in sediment samples. The highest levels of DNA damage were obtained with samples taken at and immediately downstream of urban centers. The results of this study indicate that the Noyyal River system is contaminated with substances that are genotoxic to fish and earthworms and that the comet assay has sufficient sensitivity to detect the genotoxicity.

Chromosomal aberration, micronucleus and Comet assays on peripheral blood lymphocytes of leprosy patients undergoing multidrug treatment. (Mutagenesis, 17, 309-312, 2002)

To evaluate the genetic damage in leprosy patients, we carried out the alkaline Comet assay and chromosomal aberration (CA) and micronucleus (MN) tests in peripheral blood lymphocytes of 50 leprosy patients receiving multidrug treatment (MDT) and 50 healthy individuals. The Comet assay showed statistically higher mean values for length to width ratios of DNA mass (P < 0.01) and for mean frequencies of tailed cells (P < 0.001) in cells of leprosy patients than in those of controls. Similarly, the mean frequencies of micronucleated cells (per 1000 cytochalasin B-induced binucleated cells) were significantly greater (P < 0.001) in leprosy patients (19.92 +/- 2.564) than in controls (1.6 +/- 0.231). A statistically significant 10-fold increase in the frequency of CAs (11.16 +/- 0.411) was observed in leprosy patients compared with controls (1.28 +/- 0.242). In multiple regression analyses, when patients and controls were considered together, disease factor alone significantly influenced the genotoxicity markers. In the control group, age and alcohol consumption significantly influenced MN and length to width ratios and CA frequency, respectively. However, in MDT-treated leprosy patients none of the other confounding factors (sex, age, smoking and alcohol drinking) significantly affected the extent of genetic damage.

Chromosomal aberration, micronucleus and Comet assays on peripheral blood lymphocytes of leprosy patients undergoing multidrug treatment.

To evaluate the genetic damage in leprosy patients, we carried out the alkaline Comet assay and chromosomal aberration (CA) and micronucleus (MN) tests in peripheral blood lymphocytes of 50 leprosy patients receiving multidrug treatment (MDT) and 50 healthy individuals. The Comet assay showed statistically higher mean values for length to width ratios of DNA mass (P < 0.01) and for mean frequencies of tailed cells (P < 0.001) in cells of leprosy patients than in those of controls. Similarly, the mean frequencies of micronucleated cells (per 1000 cytochalasin B-induced binucleated cells) were significantly greater (P < 0.001) in leprosy patients (19.92 +/- 2.564) than in controls (1.6 +/- 0.231). A statistically significant 10-fold increase in the frequency of CAs (11.16 +/- 0.411) was observed in leprosy patients compared with controls (1.28 +/- 0.242). In multiple regression analyses, when patients and controls were considered together, disease factor alone significantly influenced the genotoxicity markers. In the control group, age and alcohol consumption significantly influenced MN and length to width ratios and CA frequency, respectively. However, in MDT-treated leprosy patients none of the other confounding factors (sex, age, smoking and alcohol drinking) significantly affected the extent of genetic damage.

Genotoxicity evaluation of polluted ground water in human peripheral blood lymphocytes using the comet assay.

This paper presents the genotoxicity experiments with the ground water collected from an area under the influence of textile dyeing and bleaching industries in Tirupur, Tamilnadu, India. The alkaline single cell gel electrophoresis (SCGE) assay was performed in vitro with human peripheral blood lymphocytes. The cells were exposed to two doses of non-volatile organic agents extracted from ground water samples. Ground water samples were collected from 12 locations distributed in and around Tirupur and extracts were taken at different pHs (without pH adjustment and acidic pH 2.0). The persistence of the DNA damage after exposure to the organic extracts was also studied. All the samples were found to contain substances capable of inducing DNA damage in human lymphocytes. Extracts from acidified waters (pH=2.0) were found to induce more DNA damage than extracts from without pH adjustment (natural pH). The DNA damage was not fully repaired after incubation for 2h at 37 degrees C. The chemical characterization of the sub-fractions revealed the existence of aromatic amines in the extracts, which may be responsible for the DNA damaging activity of the water samples. The results of this investigation demonstrate the application of the comet assay in environmental monitoring studies.

Chromosomal aberration, micronucleus and Comet assays on peripheral blood lymphocytes of leprosy patients undergoing multidrug treatment

To evaluate the genetic damage in leprosy patients, we carried out the alkaline Comet assay and chromosomal aberration (CA) and micronucleus (MN) tests in peripheral blood lymphocytes of 50 leprosy patients receiving multidrug treatment (MDT) and 50 healthy individuals. The Comet assay showed statistically higher mean values for length to width ratios of DNA mass (P < 0.01) and for mean frequencies of tailed cells (P < 0.001) in cells of leprosy patients than in those of controls. Similarly, the mean frequencies of micronucleated cells (per 1000 cytochalasin B-induced binucleated cells) were significantly greater (P < 0.001) in leprosy patients (19.92 +/- 2.564) than in controls (1.6 +/- 0.231). A statistically significant 10-fold increase in the frequency of CAs (11.16 +/- 0.411) was observed in leprosy patients compared with controls (1.28 +/- 0.242). In multiple regression analyses, when patients and controls were considered together, disease factor alone significantly influenced the genotoxicity markers. In the control group, age and alcohol consumption significantly influenced MN and length to width ratios and CA frequency, respectively. However, in MDT-treated leprosy patients none of the other confounding factors (sex, age, smoking and alcohol drinking) significantly affected the extent of genetic damage.