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Biomedical subjects

R C Chaubey

Publications and source records attributed to R C Chaubey.

12 recordsLinked to original sources
PubMed2006

Protection against radiation oxidative damage in mice by Triphala.

Protection against whole body gamma-irradiation (WBI) of Swiss mice orally fed with Triphala (TPL), an Ayurvedic formulation, in terms of mortality of irradiated animals as well as DNA damage at cellular level has been investigated. It was found that radiation induced mortality was reduced by 60% in mice fed with TPL (1g/kg body weight/day) orally for 7 days prior to WBI at 7.5 Gy followed by post-irradiation feeding for 7 days. An increase in xanthine oxidoreductase activity and decrease in superoxide dismutase activity was observed in the intestine of mice exposed to WBI, which, however, reverted back to those levels of sham-irradiated controls, when animals were fed with TPL for 7 days prior to irradiation. These data have suggested the prevention of oxidative damage caused by whole body radiation exposure after feeding of animals with TPL. To further understand the mechanisms involved, the magnitude of DNA damage was studied by single cell gel electrophoresis (SCGE) in blood leukocytes and splenocytes obtained from either control animals or those fed with TPL for 7 days followed by irradiation. Compared to irradiated animals without administering TPL, the mean tail length was reduced about three-fold in blood leukocytes of animals fed with TPL prior to irradiation. Although, similar protection was observed in splenocytes of TPL fed animals, the magnitude of prevention of DNA damage was significantly higher than that observed in leukocytes. It has been concluded that TPL protected whole body irradiated mice and TPL induced protection was mediated through inhibition of oxidative damage in cells and organs. TPL seems to have potential to develop into a novel herbal radio-protector for practical applications.

Administration, Oral↗
PubMed2005

Computerized image analysis software for the comet assay.

Single-cell gel electrophoresis (SCGE) or the comet assay is a powerful tool for the detection of DNA single- and double-strand breaks and base damage and for investigating the kinetics of DNA strand break rejoining in human and animal model systems. It is a versatile technique that can be applied in various areas of biomedical research. This chapter highlights the importance of computerized analysis and data processing for the comet assay and describes the criteria used for manual evaluation of comets and their limitations compared with the computer-based analysis. It describes in detail SCGE-Pro, a semiautomatic software developed in our laboratory for comet evaluation and data processing. For comparison, some of the commercially available software for analysis of data from the comet assay is also described.

Animals↗
PubMed2001

Gamma ray induced DNA damage in human and mouse leucocytes measured by SCGE-Pro: a software developed for automated image analysis and data processing for Comet assay.

The studies reported in this communication had two major objectives: first to validate the in-house developed SCGE-Pro: a software developed for automated image analysis and data processing for Comet assay using human peripheral blood leucocytes exposed to radiation doses, viz. 2, 4 and 8 Gy, which are known to produce DNA/chromosome damage using alkaline Comet assay. The second objective was to investigate the effect of gamma radiation on DNA damage in mouse peripheral blood leucocytes using identical doses and experimental conditions, e.g. lyses, electrophoretic conditions and duration of electrophoresis which are known to affect tail moment (TM) and tail length (TL) of comets. Human and mouse whole blood samples were irradiated with different doses of gamma rays, e.g. 2, 4 and 8 Gy at a dose rate of 0.668Gy/min between 0 and 4 degrees C in air. After lyses, cells were electrophorased under alkaline conditions at pH 13, washed and stained with propidium iodide. Images of the cells were acquired and analyzed using in-house developed imaging software, SCGE-Pro, for Comet assay. For each comet, total fluorescence, tail fluorescence and tail length were measured. Increase in TM and TL was considered as the criteria of DNA damage. Analysis of data revealed heterogeneity in the response of leucocytes to gamma ray induced DNA damage both in human as well as in mouse. A wide variation in TM and TL was observed in control and irradiated groups of all the three donors. Data were analyzed for statistical significance using one-way ANOVA. Though a small variation in basal level of TM and TL was observed amongst human and mouse controls, the differences were not statistically significant. A dose-dependent increase in TM (P<0.001) and TL (P<0.001) was obtained at all the radiation doses (2-8 Gy) both in human and mouse leucocytes. However, there was a difference in the nature of dose response curves for human and mouse leucocytes. In human leucocytes, a linear increase in TM and TL was observed up to the highest radiation dose of 8 Gy. However, in case of mouse leucocytes, a sharp increase in TM and TL was observed only up to 4 Gy, and there after saturation ensued. In human samples, the dose response of both TM and TL showed best fits with linear model (r(TM)=0.999 and r(TL)=0.999), where as in mouse, the best fit was obtained with Sigmoid (Boltzman) model. From the present data on leucocytes with increase in TM and TL as the criteria of DNA damage, it appears that mouse is relatively more sensitive to radiation damage than humans.

Adult↗
PubMed2001

Radioadaptive response in human lymphocytes in vitro.

Exposure to low doses of radiation and/or chemicals can prime an organism to withstand the stress of a subsequent exposure to higher doses of the same agent. In the case of radiation, this phenomenon has been called radioadaptive response. Cytogenetic studies have been undertaken in human lymphocytes to investigate adaptive response (AR) to ionizing radiation, in particular to seek the role of variables such as priming dose, cell cycle stage, and age and gender of the donor. We demonstrated that pre-exposure of lymphocytes in whole blood cultures to very low doses in the range of about 1 cGy (priming or adaptive dose [AD]) reduced the frequency of micronuclei in binucleated cells induced by 100 cGy--that is, produced an AR in these cells in vitro. However, pre-exposure of cells to 10.0 cGy did not reduce the chromosomal damage (micronuclei) induced by the challenging dose (CD) of 100 cGy under the same protocol, thus exhibiting an inverse dose-response relationship. There was marked variability in the AR among the individuals investigated in the study. The extent of AR also depended on the stage of cell cycle exposed to the CD of radiation. Maximum AR was observed when CD of 100 cGy was given 4 hours after AD, 30 hours following the mitogenic stimulation of lymphocytes. The least AR was observed when CD was given 48 hours after stimulation. Interestingly, AR was also found to be dependent on the age of the donor, a decrease in AR being observed with an increasing age. No significant difference in AR was observed between male and female donors. To understand the molecular events underlying AR, protein synthesis patterns were studied in human lymphocytes subjected to mitogen, heat, or radiation stress. Heat shock (45 degrees C, for 15 min) induced 7 proteins with molecular weights ranging from 40 to 80 kDa, while treatment with phytohemagglutinin (40 microg/mL) showed induction of 2 proteins of molecular weights 38 and 48 kDa, respectively. However, exposure of human lymphocyte cultures to gamma radiation did not significantly induce synthesis of any protein, up to 800 cGy dose. Lack of induction of proteins by gamma radiation in human lymphocytes contrasts with the previous reports showing X-ray radiation-enhanced gene expression in melanoma cells and/or human tumor fibroblasts.

Adaptation, Physiological↗
PubMed1999

Inhibition of radiation-induced DNA damage in plasmid pBR322 by chlorophyllin and possible mechanism(s) of action.

Naturally occurring compounds capable of protecting DNA against ionizing radiation and chemical mutagens have considerable potential for prevention of mutation-based health impairment including cancer and other degenerative diseases. Chlorophyllin (CHL), a water-soluble derivative of chlorophyll, has been examined for its ability to protect DNA against radiation induced strand breaks using an in vitro plasmid DNA system. Gamma-radiation, up to a dose of 6 Gy (dose rate 1.25 Gy/min), induced a dose-dependent increase in single-strand breaks (ssbs) in plasmid pBR322 DNA. CHL per se did not induce, but inhibited radiation-induced ssbs in a concentration-dependent manner; 500 microM giving about 90% protection. The protection afforded by CHL was comparatively less than that of trolox, a water-soluble analogue of alpha-tocopherol. To elucidate the underlying mechanism(s), reaction of CHL with the radiation-derived hydroxyl radical (.OH) and deoxyribose peroxyl radical (ROO.) was studied by pulse radiolysis. CHL exhibited a rate constant of 6.1+/-0.4x109 M-1 s-1 with.OH and 5.0+/-1.3x107 M-1 s-1 with ROO. To our knowledge, this is the first report providing direct evidence of free radical-scavenging properties of CHL. The results showed that CHL, effectively protects plasmid DNA against ionizing radiation, in an in vitro system independent of DNA repair or other cellular defense mechanisms. The ability of CHL to scavenge. OH and ROO., may contribute to its protective effects against radiation induced DNA damage in the pBR322 system.

Antimutagenic Agents↗
PubMed1996

Radio-adaptive response in human lymphocytes in vitro.

An attempt has been made to investigate the adaptive response to ionizing radiation in the human lymphocytes in vitro using cytochalasin-B blocked micronucleated binucleate cells (mn-BNCs) as a cytogenetic end point. Whole blood samples drawn from healthy donors, of either sex were irradiated in vitro at a dose of 1 cGy (adaptive or conditioning dose) Cobalt-60 gamma radiation (dose rate 1.12 cGy/min) at about 26 hr after mitogenic stimulation. After 31 hr of their initiation, groups of cultures were subsequently exposed to a challenging dose of 100 cGy gamma radiation (dose rate 82 cGy/min.). Eight males in the age group ranging from 25 to 55 years and eight females (age group 25 to 29 years), have been analysed during this study. Analysis of data revealed 40.6% reduction in the frequency of mn-BNCs among the males with a range from 25.7% to 54.7%. In case of females, also the per cent reduction varied from 26.3% to 49.0%, with a mean value of 33.7%. Pooling the data from males and females gave an overall reduction of 37.1% in the frequency of radiation induced mn-BNCs due to pre-exposure to 1 cGy radiation.

Adult↗
PubMed1993

Studies on the migration of micronucleated erythrocytes from bone marrow to the peripheral blood in irradiated Swiss mice.

Micronucleated polychromatic (mn-PCE) and normochromatic erythrocytes (nm-NCE) were enumerated in the bone marrow and peripheral blood of Swiss male mice at different time intervals following whole-body (1.0 Gy) gamma-irradiation. Polychromatic cells migrated to the peripheral blood soon after their formation in the bone marrow and mn-PCE achieved a frequency close to that of the bone marrow with a delay of about 12 h. The optimal time for peripheral sampling was found to be about 36 h after radiation exposure. The frequency of mn-NCE in bone marrow and peripheral blood showed only a moderate and gradual increase till 60 h, and was much lower in the latter. In another experiment, mice irradiated with 0.42 Gy gamma-rays (0.21 Gy/h) once a day for 5, 10 or 15 days (5 days per week) showed a cumulative dose-dependent increase in the levels of mn-NCE in the peripheral blood, sampled at 7 or 21 days after the last exposure. These observations demonstrate persistence and accumulation of mn-PCE in the peripheral blood of mice during repeated exposure to ionizing radiation, and the sampling could be delayed up to several days after the last exposure. Thus, peripheral mn-PCE, scored between 24-48 h following irradiation, can be conveniently used to measure acute chromosomal damage induced by ionizing radiation in the bone marrow erythroblasts of mice, while peripheral mn-NCE are suited to monitor accumulated damage during chronic/repeated exposure.

Animals↗
PubMed1978

The effect of hycanthone and maleic hydrazide on the frequency of micronuclei in the bone-marrow erythrocytes of mice.

Male Swiss mice were assigned to 6 groups of either 3 or 4 animals each. 3 groups were given hycanthone methanesulfonate intraperitoneally, at 40, 80 or 120 mg/kg, respectively; the dose was repeated after an interval of about 24 h. At the same time 2 groups received maleic hydrazide at 100 or 200 mg/kg, and the remaining group was given dimethyl sulfoxide which was used as a solvent for both drugs. 6 h after the second injection, the mice were killed and bonemarrow preparations were made. Hycanthone induced a significant increase in the frequency of micronuclei in the polychromatic erythrocytes and suppressed the P/N ratio significantly. However, there was no dose-response relationship. Maleic hydrazide, on the other hand, failed to influence the incidence of micronuclei or the ratio of poly- to normo-chromatic erythrocytes.

Animals↗