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Biomedical subjects

B S Rao

Publications and source records attributed to B S Rao.

At least 19 recordsLinked to original sources

Conventional slow freezing, vitrification and open pulled straw (OPS) vitrification of rabbit embryos.

Three different methods of cryopreservation viz., conventional slow freezing, vitrification and open pulled straw vitrification were compared for their ability to support post thaw in vitro and in vivo development of rabbit embryos. Morula stage rabbit embryos were collected from super-ovulated donor does. They were randomly allocated to different freezing methods and stored up to 3 months in liquid nitrogen. After thawing and removal of cryoprotectants, embryos exhibiting intact zona pellucida and uniform blastomeres were considered suitable for in vitro culture and/or transfer. Three to five cryopreserved embryos placed in approximately 1 ml of culture medium (TCM 199 supplemented with foetal calf serum and antibiotics) were incubated for up to 72 h under humidified atmosphere of 5% CO2 in air at 39 degrees C. Development to hatched blastocyst stage was considered the initial indicator of success of cryopreservation of embryos. Of the embryos cryopreserved by programmed freezing, open pulled straw vitrification, vitrification-55 h pc and vitrification-72 h pc 55, 71, 17 and 48%, respectively, developed into hatched blastocysts. Similarly 19, 29, and 4% of embryos cryopreserved by programmed freezing, open pulled straw vitrification and vitrification -72 h pc developed into live offspring on transfer to recipient does. This is the first report on open pulled straw vitrification of rabbit embryos. Present results, suggest that (a) open pulled straw vitrification supports better in vitro survival of frozen thawed rabbit morulae; (b) both programmed freezing and OPS are similar but superior to vitirification in supporting in vivo survival of frozen thawed rabbit embryos.

Animals↗

Dose rate effect on micronuclei induction in cytokinesis blocked human peripheral blood lymphocytes.

The effect of dose rate on the induction of micronuclei (MN) in human peripheral blood lymphocytes was investigated over a range of dose rates from 0.125 Gy h(-1) to 178.2 Gy h(-1). The response of MN induction was fitted with a linear quadratic model and the alpha and beta coefficients were estimated. It was found that beta values decrease with decreasing dose rate as in the case of chromosomal aberration. At the dose rate of 0.125 Gy h(-1), pure linear response of MN induction was observed. An attempt was made to simulate the calibration curve for the purpose of biological dosimetry at different dose rates and exposure times. The yields when simulated with the exposure time or the dose rate are in agreement with experimental results.

Cell Division↗

The COG database: new developments in phylogenetic classification of proteins from complete genomes.

The database of Clusters of Orthologous Groups of proteins (COGs), which represents an attempt on a phylogenetic classification of the proteins encoded in complete genomes, currently consists of 2791 COGs including 45 350 proteins from 30 genomes of bacteria, archaea and the yeast Saccharomyces cerevisiae (http://www.ncbi.nlm.nih. gov/COG). In addition, a supplement to the COGs is available, in which proteins encoded in the genomes of two multicellular eukaryotes, the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster, and shared with bacteria and/or archaea were included. The new features added to the COG database include information pages with structural and functional details on each COG and literature references, improvements of the COGNITOR program that is used to fit new proteins into the COGs, and classification of genomes and COGs constructed by using principal component analysis.

Animals↗

Synthesis of some halogen-containing 1,2,4-triazolo-1,3,4-thiadiazines and their antibacterial and anticancer screening studies--part I.

A series of 7-arylidene-6-(2,4-dichloro-5-fluorophenyl)-3-substituted-1,2,4-triazolo[3,4-b]-1,3,4-thiadiazines (3) were prepared by the condensation of 4-amino-5-mercapto-3-substituted-1,2,4-triazoles (1) and 3-aryl-1-(2,4-dichloro-5-fluorophenyl)-2-bromo-2-propen-1-one (2). An alternative route for the synthesis of the title compound 3 has been described. The newly synthesised compounds were characterised on the basis of N-analyses, IR, 1H NMR and mass spectral data. Some of the newly synthesised compounds were tested for their antibacterial activities against Gram + ve and Gram - ve bacteria. Among the tested compounds 3n showed the highest degree of antibacterial activity against S. aureus and evaluation of the LD50 value of this compound was carried out. Some of the newly synthesised compounds were also screened for their anticancer activities. Among these, compounds 3b, 3g, 3n and 3p are found to be active against NCI-H460 (lung), MCF7 (breast), SF 268 (CNS) in the preliminary anticancer screening studies. Further, 60-cell-line anticancer studies of these compounds were carried out. The results of such studies are discussed in this paper.

Animals↗

Chromosome aberration analysis in radiotherapy patients and simulated partial body exposures: biological dosimetry for non-uniform exposures.

Chromosome aberration analysis was carried out in peripheral blood lymphocytes of cancer patients following radiotherapy of lungs, cervix and spine. Radiotherapy in the pelvic region involving large doses (6 Gy) showed an overdispersed distribution of dicentrics. However, when the doses were fractionated (three fractions of 2 Gy) distribution was found to be near Poisson. Spine irradiation covering almost all the lymphocytes pools, indicated a Poisson distribution. The data show that depending on the sites of exposure, the distribution of dicentrics in cells varies and hence there is a non-uniform distribution of lymphocytes in the body. The average dose to the lymphocytes was found to be one sixth of the partial body dose. Based on the non-Poisson distribution of aberrations, the fraction of lymphocytes irradiated, mean dose to the fraction and part of the body exposed was calculated in a case of acute 6 Gy pelvic irradiation. The fraction of cells irradiated was calculated to be 4.11% and the portion of the body exposed was approximately 16.8%. The dose to the irradiated fraction was found to be 5.4 Gy, which is in agreement with the given dose of 6 Gy. In simulated exposures the u values increased systematically with the decrease in fraction of irradiated cells and the calculated dose to the fraction was also in good agreement with the true dose.

Chromosome Aberrations↗

Retrospective biological dosimetry of absorbed radiation.

Frequencies of chromosomal translocations in human peripheral blood lymphocytes irradiated in vitro by 200 kV X rays have been estimated by the fluorescence in situ hybridisation (FISH) technique. Probes specific for whole chromosomes 1, 3 and 4 were labelled with, biotin 16 dUTP + digoxigenin 11 dUTP; biotin 16 dUTP and digoxigenin 11 dUTP respectively, to achieve three colour FISH to distinguish exchanges involving individual chromosomes. The translocation frequencies for individual chromosomes suggested a relationship between the response and the genomic content of the chromosomes studied. Dose-response curves derived from translocations involving all the three painted chromosomes (22.3% of the genome) showed a linear-quadratic response with alpha and beta coefficients of 0.027 +/- 0.009 Gy-1 and 0.033 +/- 0.004 Gy-2, corresponding coefficients for the response curves extrapolated to the entire genome are 0.072 +/- 0.026 Gy-1 and 0.075 +/- 0.011 Gy-2 respectively. Dose-response curves for the induction of dicentrics involving painted chromosomes also showed a similar response, but the dicentric frequencies were consistently lower than the translocation frequencies for all the doses studied. Dose fractionation resulted in a small but significant reduction in the yield of exchanges, thus suggesting repair of some radiation damage during the period between the two fractions. Retrospective biological dosimetry was attempted with FISH assay of five radiation workers with chronically accumulated doses of approximately 500 mSv received over a period of 2-3 decades. Among the 2421 metaphases scored in these subjects 28 translocations involving the painted chromosomes were detected. The dose estimates based on FISH assay ranged from 0.215 +/- 0.116 Gy to 0.635 +/- 0.292 mGy, thus suggesting the usefulness of this assay in retrospective biological dosimetry.

Adult↗

Effect of caffeine on the genotoxic effects of gamma radiation and 4-NQO in diploid yeast.

Caffeine is an environmental agent to which people are commonly exposed through medicines, drinks, food items, etc. It has been shown to be mutagenic in a number of test systems. In addition, it has also been shown to modify the mutagenic response of ionizing radiation, UV, and several chemical mutagens in a number of test systems. We have studied the effect of caffeine on gamma radiation and 4-Nitroquinoline 1-oxide (4-NQO)-induced gene conversion in the yeast Saccharomyces cerevisiae D7. Stationary phase cells were either exposed to 100-600 Gy of 60Co gamma radiation or treated with 0.15-0.3 microM 4-NQO (30 degrees C, 1 hour), after which they were plated on synthetic complete or minimal media with or without caffeine. Caffeine concentrations ranged from 5 to 15 mM. The results indicated that caffeine at 5 and 10 mM decreased gamma radiation-induced gene conversion frequencies significantly at 400 and 600 Gy. At 600 Gy, the decrease was about 30% and 50% with caffeine concentrations of 5 and 10 mM, respectively. In contrast, caffeine was found to increase the induced gene conversion frequency when cells treated with 0.15, 0.225, and 0.3 microM 4-NQO were plated on media containing caffeine. The increase with 5, 10, and 15 mM caffeine was approximately 1.5, 2, and 2.5, respectively, times the value of 4-NQO alone. The results indicate that the posttreatment repair processes following gamma irradiation or 4-NQO treatment are modified via different pathways.

4-Nitroquinoline-1-oxide↗

Retrospective dosimetry based on cytogenetic study of individuals chronically exposed to ionizing radiation.

Analysis of chromosomal aberrations (CAA) was carried out in 27 radiation workers with cumulative exposures of approximately 500 mSv received over a period of 2-3 decades. A similar study was carried out in 20 age- and gender-matched healthy individuals who served as controls. The average age of the exposed group was 50 years, and that of the controls was 51 years. The absorbed radiation dose was calculated by using the linear component of the in vitro dose response curve established for 60Co gamma-rays. In the controls we found only 3 dicentrics in 5,500 metaphases analyzed. In the exposed group, we detected 49 dicentrics and 1 centric ring in 13,900 metaphases analyzed. Because of the small number of dicentrics scored in each individual, the dose estimates of our method of study suffers from a large statistical uncertainty. The collective dose calculated from our data was 3.4 Sv. The difference in the physical dose based on personal monitoring and that from biological dosimetry may be attributed to the disappearance of lymphocytes carrying the aberrations from the living system. These results are consistent with the present knowledge of the kinetics of turnover of T-lymphocytes. Most residual damage appears to be from the long-lived component of T-lymphocytes with a mean life of 10 years.

Air Pollution, Radioactive↗

Synthesis of some new biologically active bis-(thiadiazolotriazines) and bis-(thiadiazolotriazinyl) alkanes.

4-Amino-3-mercapto-1,2,4-triazin-5(4H)-ones (1) were condensed with dicarboxylic acids 2 to yield bis-(4-oxo-4H-1,3,4-thiadiazolo[2,3-c]-1,2,4-triazin-7-yl)alkanes (3b-d,f-h,j-l,n-p) and bis-thiadiazolotriazines (3a,e,i,m). All the newly synthesised compounds were characterised by analytical, IR, NMR and mass spectral studies. Some of the newly synthesised compounds were screened for their antibacterial and antifungal properties. Among the tested compounds, compound 7,7'-(1,4-butanediyl)-his-(3-t-butyl-4-oxo-4H-1,3,4-thia-diazolo[2,3-c]-1,2,4-triazine (3p) exhibited highest degree of antifungal activity.

Alkanes↗

Chromium removal and reduction in COD of tannery effluents by actinomycetes.

Tannery effluents are highly polluting and contain chroomium and high Cod and Bod. Alkalotolerant/alkalophilic actinomycetes NCIM 5080 and NCIM 5142 have been shown earlier to tolerate and accumulate chromium during growth also produce alkaline protease in presence of chromium ions, these properties of the isolates are suitable for treatment of tannery effluents which are alkaline and contain chromium and proteinaceous matter, both the actinomycetes are able to grow in undiluted tannery effluents and remove chromium almost completely and reduce the COD by 70%-80% during growth as well as by pregrown biomass.

Actinobacteria↗

Studies on arylfuran derivatives. Part XI. Synthesis, characterisation and biological studies on some Mannich bases carrying 2,4-dichlorophenylfurfural moiety.

A series of 3-substituted-4-[5-(2,4-dichlorophenyl)-2-furfurylidine]amino-5-me rcapto-1, 2,4-triazoles (3) are synthesised. Aminomethylation of 3 with formaldehyde and a primary/secondary amine furnished Mannich bases 4 and 5. Both Schiff bases 3 and Mannich bases 4 and 5 are characterised on the basis of IR, 1H NMR, mass spectral data and elemental analysis. All the newly synthesised compounds are tested for their antibacterial activities. Some of the selected compounds are also tested for their fungicidal and herbicidal properties.

Anti-Infective Agents↗

Restraint stress-induced alterations in the levels of biogenic amines, amino acids, and AChE activity in the hippocampus.

Inspite of large number of studies on the neurochemical changes in the stress, an equivocal case is yet to be made for the role of a specific neurotransmitter in this important neurobiological disorder. The difficulty arises from the fact that there is no single neurotransmitter system appears to be responsible for the stress induced damage to the hippocampal neurons. The present study evaluates the effect of restraint stress on the alterations in the levels of biogenic amines, aminoacids and acetylcholinesterase activity in the hippocampus. Male Wistar rats of 45 days old were subjected to 6 hours of daily restraint stress over a period of 21 days. Immediately after the last session of stress, rats were sacrificed and neurotransmitter levels were estimated in the hippocampus. A significant (p < 0.001) decrease in the levels of noradrenaline, dopamine, 5-hydroxytryptamine and acetylcholinesterase activity in the stressed rats was observed compared to controls. However, levels of glutamate was significantly (p < 0.001) increased in stressed rats. These results indicate that chronic restraint stress decreases aminergic and cholinergic neurotransmission, and increases the glutamatergic transmission in the hippocampus.

Acetylcholinesterase↗

Preparation and in vitro evaluation of chitosan matrices cross-linked by formaldehyde vapors.

Rifampicin-chitosan matrices were prepared by a chemical cross-linking method to develop a sustained-release form. The effects of cross-linking agent (formaldehyde) on the drug release rate and release kinetics were investigated in this study. Moreover, the kinetics of rifampicin released from chitosan matrices exposed to formaldehyde vapors for predetermined time intervals were analyzed using Ritger and Peppas exponential equation. The in vitro release kinetics exhibited a non-Fickian transport model. Increasing the exposure time to formaldehyde vapors decreased the release rate of rifampicin from chitosan matrices as a result of formation of greater structural strength and tighter texture.

Antibiotics, Antitubercular↗

A double blind controlled study of propranolol and cyproheptadine in migraine prophylaxis.

Role of propranolol and cyproheptadine in the prophylaxis of migraine was studied in a controlled double blind trial. Two hundred fifty-nine patients were divided into four groups. Each group was either given a placebo, cyproheptadine, propranolol or a combination of the latter two drugs. The patients were followed for a period of three months. Significant relief in frequency, duration and severity from migranous attacks was seen in all drug treated groups over placebo. Significant correlation in response was seen in frequency, duration and severity in all the groups which received drugs. Statistically more significant relief was seen in cyproheptadine and propranolol treated group as compared to individual drug treated groups. In cyproheptadine and propranolol treated groups, the dropout rate was lower and associated symptoms were better relieved than in other groups. The study shows efficacy of combination of cyproheptadine and propranolol in migraine prophylaxis.

Adolescent↗

Radiosensitization of a mouse melanoma by withaferin A: in vivo studies.

The radiosensitizing effect of a plant withanolide, withaferin A, on the B16F1 mouse melanoma was studied in vivo. Treatment of 100 mm3 tumours with 10 to 60 mg/kg withaferin A intraperitoneally produced a dose dependent increase in growth delay and volume doubling time. Injection of 30-50 mg/kg withaferin A, followed by 30 Gy local gamma irradiation, significantly enhanced the tumour response. No systemic or local adverse reactions were noted in these groups. The drug was most effective when injected intraperitoneally 1 h before irradiation. However, neither the individual agents nor their combination could produce any complete response (tumour cure). Melanoma is a relatively radioresistant tumour. The present results indicate that the radiation response of this tumour can be significantly enhanced by pretreatment with withaferin A.

Animals↗

Increased numerical density of synapses in CA3 region of hippocampus and molecular layer of motor cortex after self-stimulation rewarding experience.

Self-stimulation has been considered as an intensely rewarding behavioural experience, being perhaps even more influential than feeding or sexual behaviour. Our earlier studies have demonstrated a self-stimulation rewarding experience-induced increase in dendritic branching points, intersections and spine densities in CA3 hippocampal and layer V motor cortical pyramidal neurons. In the present study, we report self-stimulation-induced alterations in the numerical density of synapses in the hippocampus and motor cortex. A self-stimulation experience was provided 1 h daily for a period of 10 days through bipolar electrodes, implanted bilaterally in the lateral hypothalamus and substantia nigra-ventral tegmental area, stereotaxically. The results revealed a significant (P < 0.001) increase in the number of synapses in the CA3 region of hippocampus and the molecular layer of the motor cortex in self-stimulation-experienced rats. The increased synaptic number may be due to the activation of afferent pathways to the hippocampus and motor cortex following self-stimulation, which may lead to the induction of long-term potentiation. Long-term potentiation is known to cause structural changes by strengthening the existing synapses or resulting in the formation of new synapses. These changes may be related to the improved cognitive functions observed in self-stimulation-experienced rats.

Animals↗