Yogendra Verma

Association between sperm quality, oxidative stress, and seminal antioxidant activity.

OBJECTIVES To determine seminal antioxidant capacity, oxidative stress markers, and their association with semen quality as oxidative stress is considered to be a major etiological factor in male infertility. SUBJECTS AND METHODS Semen samples were obtained from 138 men and categorized on the basis of sperm count, motility, and morphology. Seminal oxidative and antioxidant markers are as follows: lipid peroxidation (LPO), protein carbonyls (PC), superoxide dismutase (SOD), catalase (CAT), thiols, and ascorbic acid were determined. RESULTS Sperm count significantly correlated positively with progressive sperm motility and normal morphology. Sperm count and normal morphology showed significant negative correlation with LPO and PC. Sperm count and progressive motility showed significant positive relationship with SOD. The SOD, CAT, and thiols positively whereas LPO and PC negatively associated with elevated sperm count. CONCLUSION Insufficient antioxidant enzymes and increased oxidative stress may attribute to the risk of declining semen quality and hence protective role for antioxidant enzymes against the oxidative damage cannot be ruled out.

Effect of cadmium on blood of Tilapia,Oreochromis mossambicus (peters), during prolonged exposure

Cadmium is recognized as one of the most hazardous environmental pollutants and is toxic to many living organisms. Experimental and environmental exposure to cadmium has been reported to cause disease in humans and other mammals. Recent reviews on cadmium have reported on acute and subacute effects on fish, mechanisms of toxicity, the role of toxicity modifying factors and various sublethal effects, i.e., hematological and biochemical disorders. However, very little information is available on the subacute effects of cadmium on fish blood exposed in hard water. The present investigation, therefore, was carried out to observe the effects of cadmium on the blood of tilapia (Oreochromis mossambicus) in hard and alkaline water to determine whether cadmium causes changes in blood of fish kept in hard water in the same way it causes changes in blood of fish kept in soft water.

Cytogenetic alterations in buccal mucosa cells of chewers of areca nut and tobacco

OBJECTIVE the rationale of the study was to evaluate the cytological alterations especially micronucleus (MN) and other nuclear anomalies in buccal mucosa cells of chewers to understand the genotoxic and clastogenic potential of chewing mixture (containing areca nut and tobacco as main ingredients). METHODS the buccal cytome assay involves the examination of epithelial smear to determine micronucleated cell and other nuclear anomalies after the Feulgen plus light green staining. The assay was applied to exfoliated buccal mucosa cells of 262 subjects [non-chewers - 161 and chewers - 101 (includes 20 subjects with OSMF)] and 1000 cells per individual were examined microscopically. Nuclear anomalies were compared among chewers, non-chewers and OSMF subjects and correlated with consumption of quids per day and duration of chewing in years. RESULTS MN cells were found significantly (p<0.0001) higher among chewers and OSMF subjects as compared to non-chewers. Further analysis indicated that MN was significantly higher in OSMF subjects with respect to even chewers. Nuclear buds were significantly higher (p<0.0001) in OSMF subjects as compared to chewers as well as non-chewers. Nuclear anomalies viz. binucleated, karyorrhexis and karyolysis were also considerably higher in OSMF subjects as compared to non-chewers. CONCLUSION the MN and other nuclear anomalies reflected genetic damage and cytotoxicity, associated with tobacco and areca nut consumption. Further, these data reveal a risk for development of OSMF among chewers of mixture containing areca nut and/or tobacco, as all the OSMF subjects were chewers.

Toxicity assessment of dye containing industrial effluents by acute toxicity test using Daphnia magna.

Toxicity of dye containing effluent of tannery, textile, dyes and pulp-paper industries was evaluated in an acute toxicity test using Daphnia magna. The 48-hour EC50 values were 4.33% and 19.5% for tannery effluents (Tn1 and Tn2). Textile effluents (Tx1—Tx7) had 48-hour EC50 values; >100%, >100%, 62.9%, 63.0%, 40.3%, >100% and >100%, respectively. Dye industries (D1—D7) had 48-hour EC50 values; 14.1%, 15.5%, 24.5%, 29.7%, 23.2%, >100% and >100%, respectively. Similarly pulp-paper effluents (P1—P5) showed acute toxicity as 100%, 77.87%, 46.44%, 69.55% and 82.84%, respectively. These results showed linear relationship with high degree of confidence (r2 ≥ 0.84—0.99) between immobility and test concentrations. Toxicity classification criteria showed that out of five effluents from pulp-paper mill, four were minor acutely toxic having 48-hour EC50 value in between >46%—100%. Out of seven textile effluents, four were not acutely toxic (48-hour EC50 value >100%) and three were minor acutely toxic (48-hour EC 50 value in the range of 40.3%—63.0%). Similarly, out of seven dye industrial effluents, two were not acutely toxic and five minor acutely toxic. One of the two tanneries was moderately acutely toxic and another one was minor acutely toxic. Classification based on toxic unit revealed that four out of five pulp-paper effluent, three out of seven textile effluents, five out of seven dye effluents and both the tannery effluents were toxic. Overall, 66.67% effluents were found toxic and 33.33% as non-toxic. In general, tannery and dyes effluents showed more toxicity than textile and paper mill effluents.

Assessment of Genetic Damage Among Chewers of Mixture Containing Mainly Areca Nut and Tobacco

Chewing mixture containing areca nut and tobacco is believed to be associated with oral cancer. Habit of chewing such mixture is prevalent among South Asian countries. This study aimed to evaluate the genotoxic effect of areca nut and tobacco on human lymphocytes. Peripheral blood from 107 subjects (nonchewers, 48; chewers, 59, including 20 subjects with oral submucous fibrosis [OSMF]) analyzed by cytokinesis-block micronucleus (CBMN) and alkaline comet assay. Nuclear anomalies, namely, binucleated cells with micronuclei (BN MN), total MN, nucleoplasmic bridge, and nuclear buds were higher in chewers whereas elevation in BN MN and total MN were significant among subjects with OSMF than nonchewers. DNA damage assessed by comet assay showed increased percentage of Tail DNA, Tail moment, and Olive tail moment among chewers as well as OSMF subjects. Significant positive correlation was observed between induction of CBMN and consumption of quids per day (r = .280, P = .033). Results suggested cytotoxic and genotoxic potential of mixture containing areca nut and tobacco.

Evaluation of genotoxicity of pan masala employing chromosomal aberration and micronucleus assay in bone marrow cells of the mice.

Pan masala is commonly consumed in south-east Asian and other oriental countries as an alternate of tobacco chewing and smoking. Genotoxic potential of pan masala (pan masala plain and pan masala with tobacco known as gutkha) was evaluated employing chromosome aberration (CA) and micronucleus (MN) assay in vivo. Animals were exposed to three different doses (0.5%, 1.5% and 3%) of pan masala plain (PMP) and gutkha (PMT) through feed for a period of 6 months and micronucleus and chromosomal aberrations were studied in the bone marrow cells. Induction of mean micronuclei in polychromatic erythrocytes (MNPCE) and normochromatic erythrocyte (MNNCE) was higher in both types of pan masala treated groups with respect to control group. Both pan masala plain and gutkha treatment significantly induced the frequency of MNPCE and MNNCE in the bone marrow cells, indicating the genotoxic potential. Furthermore, slight decline in the ratio of polychromatic erythrocytes to normochromatic erythrocytes was also noticed, suggesting the cytotoxic potential even though the ratio was statistically non significant. A dose-dependent, significant increase in chromosome aberration was observed in both types of pan masala treated mice with respect to control. However, no significant difference in micronucleus and chromosomal aberration induction was noticed between two types of pan masala exposed (PMP and PMT) groups. Results suggest that both types of pan masala, i.e. plain and gutkha, have genotoxic potential.

Lead-induced biochemical changes in freshwater fish Oreochromis mossambicus

Lead, a non-essential and non-beneficial element has considerably added the problem of health hazard to human and experimental mammals. It has also received much attention over the past few years as potentially important aquatic pollutant. Fishes are of great nutritional significance and their intoxication by lead causes retardation of growth and deterioration in the nutritional value. Very little attention has been paid to biochemical changes which develop more quickly in response to toxicants than any apparent morphological changes. Therefore, the present investigation was undertaken to evaluate the effect of lead on plasma chemistry of freshwater fish Oreochromis mossambicus. This fish was selected because of its wide availability, edibility in India and its suitability as a model fish for toxicity testing. The variables such as glucose, cholesterol and protein representing carbohydrate, lipid and protein metabolism were studied.

Assessment of estrogenic potential of di-n-butyl phthalate and butyl benzyl phthalate in vivo

Phthalate compounds are widely used industrial chemicals; when incorporated into polyvinyl chloride, they are not covalently bound and released into the surrounding media. Some of them have estrogenic potential in vitro but data on in vivo studies are scanty. For the 3-day uterotrophic assay, di-n-butyl phthalate (DBP;10 and 100 mg/kg), butyl benzyl phthalate (BBP; 20 and 200 mg/kg), and diethylstilbestrol (DES, 40 µg/kg, positive control) were administered orally to immature female rats for three consecutive days from postnatal day (PND) 21. For the 20-day pubertal onset assay, DBP (10 and 20 mg/kg), BBP (20 and 200 mg/kg), and DES (6 µg/kg) were administered orally from PND 21 daily for 20 days. In the uterotrophic assay, in groups treated with higher dose of DBP and BBP, the uterine wet weight significantly decreased in the higher dose, and there were minor variations in the ovary wet weight, while the wet weight of these organs increased significantly in DES-treated group. In the 20-day pubertal assay, the weight of uterus and ovary declined significantly and changes in vaginal weight were nonsignificant in DBP- and BBP-treated groups. However, in DES-treated group nonsignificant elevation in vagina weight was observed. All the DES-treated animals showed the vaginal opening (VO) on day 26.17 ± 0.16. However, VO was not observed in any of the animals in control, vehicle control, BBP-, and DBP-treated groups up to PND 42, except in one animal each in vehicle control and DBP (100 mg/kg)-treated groups. The data indicated that both DBP and BBP were unable to induce elevation in the uterine and ovarian weight. While DES treatment can accelerate the growth of uterus and ovary and alter the onset of puberty and estrous cyclicity in prepubertal rats. These suggest that these compounds may not have estrogenic potential in vivo.

Reproductive toxic potential of panmasala in male Swiss albino mice

Some ingredients of panmasala have the ability to penetrate the blood-testis barrier but the reproductive toxic potential of panmasala has not been studied. This study is aimed to assess the possible damage caused by panmasala to male reproductive system in mice. Swiss albino male mice were randomly divided into 7 groups receiving either standard control diet or panmasala-containing diet. Three doses (0.5%, 1.5% and 3%) of panmasala plain (PMP) as well as panmasala with tobacco (PMT)—gutkha were given for a period of 6 months. Assessment of organ weight, sperm count and morphology, spermatid count, sperm production, testicular 17β-hydroxysteroid dehydrogenase (17β-HSD) activity and histology were conducted. A nonsignificant decrease in absolute and relative weight of testis and epididymis was observed. Spermatid count, sperm count and production were significantly decreased and 17β-HSD activity was found considerably declined at 3% of both PMP- and PMT-treated groups as compared to control. The histological observations revealed panmasala induced testicular damage. Abnormal morphology of sperm head shape was significantly elevated in higher doses of both types of panmasala-treated groups than control. The results suggests that panmasala has reproductive toxic potential and more alteration is seen with gutkha as compared to panmasala plain, indicating that similar effects might also be possible in humans.