Sequential effects of ultraviolet radiation on the histomorphology, cell density and antioxidative status of the lens epithelium--an in vivo study.

In vivo progressive effects of UV irradiation on the lens epithelium were studied using various histomorphological and biochemical parameters. Fifteen day old rat pups were exposed to 600 mW/m2 of radiation, including UV-A and UV-B, 12 h daily for 90, 120, 150 and 180 days. Biochemical parameters such as protein-bound and non-protein-bound sulfhydryl groups in both soluble and insoluble fractions and enzymes, which play an important role in combating the oxidative stress, were studied. Decreased cell density of lens epithelial cells (LEC) was observed in all three zones along with the decrease in the levels of soluble sulfhydryls (S-SH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). On the other hand, an increase in insoluble sulfhydryls was observed. Because of the decrease in S-SH and GR activities, the LEC became vulnerable to oxidative stress. Decreased activities of SOD, GPx and CAT suggest elevated oxidative stress. This effect of UV radiation may lead to cell death that may be responsible for the observed decrease in the cell density in all three zones of the lens epithelium.

Hepatoprotective and antioxidant property of Andrographis paniculata (Nees) in BHC induced liver damage in mice.

Andrographis paniculata (AP) treatment prevents BHC induced increase in the activities of enzymes y-Glutamyl transpeptidase, glutathione-S-transferase and lipid peroxidation. The activities of antioxidant enzymes like superoxide dismutase, catalase, glutathione peroxidase and the levels of glutathione were decreased following BHC effect. Administration of AP showed protective effects in the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase as well the level of glutathione. The activity of lipid peroxidase was also decreased. The result indicate antioxidant and hepatoprotective action of A. paniculata.

Chromosomal aberrations in young cancer patients.

Spontaneous level of chromosomal aberrations (CA) is considered to be indicative of inherent cancer predisposition, which plays a major role in total cancer incidence. We have studied spontaneous CA levels in in vitro cultured peripheral blood lymphocytes of pediatric cancer patients (n = 77). Results were compared with those of control subjects (n = 72), including: age-matched controls; elder controls (minimum age 60 years); and healthy first-degree relatives (FDR) of pediatric cancer patients. Pediatric cancer patients showed the highest mean CA/cell value, which was statistically significant as compared to their age-matched counterparts, elder controls, and the FDRs. As compared to 7% of all the three control groups collectively, 32.4% of pediatric cancer patients showed > 0.1 mean CA/cell value. One of the FDRs with a very high frequency of CA developed cancer within three years. The results suggest that spontaneous levels of chromosomal aberrations may be used as one of the biomarkers for cancer predisposition study.

Influence of UV-irradiation on enzymes in mouse ocular lens: in vitro studies.

PURPOSE: In vitro study of the enzymes involved in aerobic, anaerobic and hexose monophosphate shunt in ultraviolet radiation exposed mice lenses. METHOD: Of the selected enzymes, lactic dehydrogenase (LDH) was representative of anaerobic glucose oxidation, succinic dehydrogenase (SDH) of the aerobic oxidation, and Glucose-6-phosphate dehydrogenase (G-6-PDH) of the Hexose Monophosphate (HMP) shunt. Other enzymes studied were ATPase and glutathione reductase (GR). RESULTS: Experiments with mice lenses in vitro showed that transparent lens became opaque following UV-irradiation at 360 nm. Opacification of the lens was accompanied by a change in enzyme activities for energy metabolism. CONCLUSION: These changes were progressive in a manner analogous to sequential morphological changes, which would be crucial in maintaining lens transparency.

Mitomycin C induced chromosomal aberrations in young cancer patients.

Mitomycin-C (MMC) induced Chromosomal aberration (CA) frequencies were studied in 48 h peripheral blood lymphocyte (PBL) cultures of untreated cancer patients of young age (maximum age 12 years, n=77). Control population (n=71) consisted of age-matched group (maximum age 12 years, n=21); elder controls (minimum age 60 years, n=19) and healthy first degree relatives, i.e., parents or siblings of the pediatric cancer patients (mean age 24.3 years, n=31) as they share their genome and environment. Induced CA levels were found to be significantly higher among pediatric cancer patients as compared to control groups. The age-matched and elder control groups showed comparable CA levels. The first degree relatives controls showed higher induced CA levels as compared to pediatric and elder control groups. The present results indicate that there are different degrees of mutagen sensitivity prevailing in normal population. This may be responsible for differential cancer proneness. High degree of mutagen sensitivity in cancer patients may also be playing a major role in cancer onset at an early age.

Human lens epithelial layer in cortical cataract.

Normal and cataractous human eye lenses were studied by morphology and protein analysis. A marked decrease in protein sulfhydryl (PSH) and nonprotein sulfhydryl (NSPH) was observed in nuclear and cortical cataractous epithelia. Moreover, decrease in PSH contents and an increase in insoluble proteins were found to be correlated only in cortical cataractous epithelium which is also accompanied by various morphological abnormalities. In nuclear cataractous epithelium, however, there was very little insolubilisation of proteins. The epithelial morphology in nuclear cataracts was almost similar to normal lens epithelium. Hence, it is assumed that the protein insolubilisation and various morphological abnormalities are characteristics of cortical cataractous epithelium. This leads us to believe that opacification in cortical cataract might initiate in the epithelial layer.

Evaluation of chemopreventive effects of betel leaf on the genotoxicity of pan masala.

The antigenotoxic effect of the aqueous extract of betel leaf (BL-ext.) against the pan masala was tested with the help of cytogenetic endpoints like chromosome aberration (CA) and sister chromatid exchange (SCE) utilizing Chinese hamster ovary (CHO) cells. Compared to the cultures treated with aqueous extract of pan masala alone, a reduction in CA and SCE frequencies in CHO cells was observed following a combined treatment with pan masala (with or without tobacco) extract and BL-ext. The protective effect of BL-ext. against the genomic damage caused by pan masala was statistically significant only after treating the cells for a longer period.

Acid phosphatase and lipid peroxidation in human cataractous lens epithelium.

The anterior lens epithelial cells undergo a variety of degenerative and proliferative changes during cataract formation. Acid phosphatase is primarily responsible for tissue regeneration and tissue repair. The lipid hydroperoxides that are obtained by lipid peroxidation of polysaturated or unsaturated fatty acids bring about deterioration of biological membranes at cellular and tissue levels. Acid phosphatase and lipid peroxidation activities were studied on the lens epithelial cells of nuclear cataract, posterior subcapsular cataract, mature cataract, and mixed cataract. Of these, mature cataractous lens epithelium showed maximum activity for acid phosphatase (516.83 moles of p-nitrophenol released/g lens epithelium) and maximum levels of lipid peroxidation (86.29 O.D./min/g lens epithelium). In contrast, mixed cataractous lens epithelium showed minimum activity of acid phosphatase (222.61 moles of p-nitrophenol released/g lens epithelium) and minimum levels of lipid peroxidation (54.23 O.D./min/g lens epithelium). From our study, we correlated the maximum activity of acid phosphatase in mature cataractous lens epithelium with the increased areas of superimposed cells associated with the formation of mature cataract. Likewise, the maximum levels of lipid peroxidation in mature cataractous lens epithelium was correlated with increased permeability of the plasma membrane. Conversely, the minimum levels of lipid peroxidation in mixed cataractous lens epithelium makes us presume that factors other than lipid peroxidation may also account for the formation of mixed type of cataract.

Cholesterol and lipid peroxidation in 3 beta-(2-diethylaminoethoxy)- androst-5-en-17-oneHCl (U18666A) induced cataractogenesis in rats.

Decrease in cholesterol was observed in precataractous, cataractous, advance nuclear cataractous and non-cataractous lenses when 3 beta-(2-diethylaminoethoxy)-androst-5-en-17- oneHCl (U18666A) was injected, sc, to rats. Significant increase in lipid peroxidation was observed before the onset of any apparent lenticular opacity in U18666A treated rats. The results suggest that decrease in cholesterol is capable of altering the structural integrity of lens fibers. However, 12.5% decrease in cholesterol and 5% increase in lipid peroxidation observed in non-cataractous lenses indicated that these changes are not sufficient for any apparent opacification.

Lens epithelial cell density and histomorphological study in cataractous lenses.

Human epithelial cell density was determined from flat preparation of 195 cataractous lenses from 108 males and 87 females between 30 and 80 years of age. The mature cataracts had significantly lower cell counts than the other cataracts. Cell density was significantly higher in the females than in the males. Morphohistological study of the epithelia was focused on the following cataract types: (1) nuclear, (2) posterior subcapsular, (3) mature, (4) mixed, (5) hypermature, and (6) black. The major cataractous changes in all types involved vacuolization of the cytoplasm. The mature types of cataractous epithelia showed 56% superimposed cells; the epithelia in nuclear, posterior subcapsular, and black cataracts showed between 6% and 16%. In the hypermature cataracts, four of five tissues analyzed showed superimposed cells. The superimposed areas are probably the source of increased and altered cell activity. We propose that the metaplastic processes leading to posterior capsular opacification originate from these areas. The majority of nuclear and black cataracts were almost similar to the normal human lens epithelium with more or less uniform distribution of cells. Nucleus shrinkage (5 microns) was more evident in nuclear cataracts; in subcapsular cataracts most of the nuclei were large (average 9 microns diameter). Variation in morphological changes like vacuolization of cytoplasm and nuclei, pyknotic nuclei, and superimposed cells was more evident in the mixed type of cataracts.

Glutathione and glutathione-related enzymes in busulfan treated rat lens.

Glutathione (GSH) and GSH-related enzymes, glutathione reductase (GR), gamma-glutamyl cysteine synthetase (gamma-GCS), gamma-glutamyl transpeptidase (gamma-GTP), glutathione S-transferase (GST) and adenosine triphosphatase (ATPase) enzymes were analysed to study the effect of busulfan on the defence mechanisms of the lens. All these enzymes were found to increase significantly except GSH which showed only 7.9% increase as compared to controls in precataractous stage. These results affirm that busulfan is capable of evoking a response from the enzymes involved in the various pathways of GSH enabling the lens to prolong its clarity. The cataractous lenses showed significant decrease in all these parameters. Here, the impairment of the defense mechanism (GST, GR) and the total ATPase may be attributed to the cumulative action of the drug which can react with -SH groups of these enzymes, ultimately causing opacification.

C-band heterochromatin variants in individuals with neoplastic disorders: carcinoma of breast and ovary.

C-band heteromorphism was studied in 106 females, including 71 patients with breast cancer or ovarian cancer. Size heteromorphism and localization variants (inversions) were scored separately. Compared to 22.86% of controls, 70.42% of the cancer patients were heteromorphic for the size of C-band. The localization variants were observed in only 8.57% controls, whereas 42.25% of cancer patients carried such variants. Taking both the criteria together, compared to 31.43% controls, 80.28% cancer patients were C-band heteromorphic. The present study supports the reported association between presence of C-band heteromorphism and occurrence of a malignant disease.

Variability of euchromatic and heterochromatic regions of Y chromosome in two types of cancer patients.

Heteromorphism of Y chromosome was studied in head and neck cancer patients and leukemia patients. The results were compared with similar data obtained for healthy men. It was observed that, compared to the controls, mean lengths of Y chromosome were nonsignificantly higher for leukemia patients and lower for head and neck cancer patients. The euchromatic region of Y chromosome (Y-eu) remained comparable in the controls and the leukemia patients, whereas it was smaller in patients with head and neck malignancies. The heterochromatic region (Y-het) was more or less analogous in controls and head and neck cancer patients, however, it was significantly larger in patients with leukemia (P less than 0.02).

Y chromosome in leukemia patients. A study on the heteromorphic nature of its heterochromatic segment.

Possible heteromorphism of Y chromosome regions was studied in 50 leukemia patients and 50 healthy controls. The mean length of the Y chromosome was insignificantly higher in the patients as compared to the controls. No difference was observed in the size of the euchromatic region; however, the size of the heterochromatic region was significantly larger in leukemia patients as compared to the controls.

Elevated sister chromatid exchange frequencies in the lymphocytes of esophageal cancer patients.

Thirty patients with esophageal cancer and 35 healthy controls were studied for spontaneous and mutagen-induced sister chromatid exchange (SCE) frequencies and cellular kinetics in peripheral blood lymphocytes. Some of the patients and controls were tobacco consumers (TC). A mean spontaneous SCE per cell value of 8.36 obtained for the patients was higher significantly (P less than 0.001) compared with a SCE per cell value of 6.74 for the controls. It was noticed that the elevation was significant even when persons in both groups were separated and compared on the basis of tobacco consumption. Among the controls, TC had higher SCE compared with non-tobacco consumers (NTC) (P less than 0.01). Mitomycin C (MMC) was used as a mutagen. No significant difference was observed in MMC-induced rates of SCE between the patients and controls. The cellular kinetics, expressed as average generation time (AGT), also were comparable in both groups. It was concluded that either metabolic stress imposed by the tumor or some clastogenic secretion of malignant cells was responsible for elevated SCE rates in the lymphocytes of the esophageal cancer patients. The disease had no effect on mutagen-induced SCE rates or cellular kinetics in lymphocytes.

Increased frequency of sister chromatid exchanges in lymphocytes of breast cancer patients.

Spontaneous and mitomycin-C (MMC)-induced rates of sister chromatid exchanges (SCEs) were studied in peripheral blood lymphocytes (PBLs) of 40 patients with cancer of the breast and 40 healthy female volunteers as controls. Spontaneous SCE per cell value in PBL cultures was 7.72 for the breast cancer patients, which was significantly higher (p less than 0.001) than the 6.28 SCEs per cell scored for the controls. Eight of the patients were studied a second time, 3 to 6 months following surgical removal of the tumour. A significant (p less than 0.05) reduction in SCE per cell value was observed following mastectomy. MMC-induced frequencies of SCE remained comparable in patients and controls. Cellular kinetics, calculated as average generation time (AGT) from the frequency of cells in M1, M2 and M3 cycles, were comparable in patients and controls.

Enhancement of lymphocytic SCE frequencies in patients with ovarian cancer.

Sister chromatid exchange (SCE) frequencies were studied in normal and mutagen-treated lymphocyte cultures from patients with ovarian carcinoma and controls. The mean SCE value per cell in normal cultures from the patients was 7.84 which was significantly higher than 6.41 SCEs per cell observed in the controls (p less than 0.01). Similar SCE values obtained for mutagen (mitomycin C, MMC) treated cultures were somewhat lower for patients as compared to the controls, however, the difference was not significant. Cellular kinetics, expressed as average generation time (AGT), was suppressed in lymphocyte cultures from ovarian carcinoma patients as compared to the controls (p much less than 0.05). It was concluded that although baseline SCE rates in lymphocytes of patients with ovarian carcinoma were higher as compared to the controls, they have very limited application as a marker malignancy.