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The toxicity of carmoisine and of its metabolites after interaction with media components, was studied using the single cell system of Tetrahymena pyriformis. Carmoisine was not toxic to T. pyriformis. However, when autoclaved with media components such as bactopeptone or yeast extract, it gave rise to product(s) that completely inhibited the growth of T. pyriformis. Addition of glucose to the bactopeptone-carmoisine mixture before autoclaving significantly enhanced the formation of the toxic product. A similar effect was observed with other reducing sugars and with compounds containing sulfhydryl groups. The results indicate that carmoisine may undergo initial reduction, followed by a series of reactions leading to the formation of the toxic component(s).

The mutagenic potential of riboflavin and its photodegradation product lumiflavin was evaluated using the umu test, SOS chromotest and Ames Salmonella assay. Both riboflavin and lumiflavin by themselves were found to be non-mutagenic. On treatment with rat liver microsomal enzymes (S9) or caecal cell-free extract (CCE), lumiflavin acquired mutagenicity, while the status of riboflavin remained unaffected. Activation of lumiflavin by metabolic enzymes was found to result in an alteration of its spectral characteristics.

On exposure to visible light, riboflavin and lumiflavin produced reactive oxygen species such as singlet oxygen and superoxide radicals. The reaction was found to be time- and concentration-dependent. Both riboflavin and lumiflavin, upon illumination, showed mutagenic response in the umu test as well as in the Ames/Salmonella assay with Salmonella typhimurium TA102. The mutagenic response was partially abolished by superoxide dismutase while sodium azide did not have any effect. No mutagenicity was observed if the compounds were not illuminated. The results suggested the involvement of superoxide radicals in light-induced mutagenicity of riboflavin as well as lumiflavin.

Whole-body exposure of rats to 1000 R X-rays resulted in decreased rates in protein and RNA syntheses in the spleen from the fourth post-irradiation hour. These changes correspond well with impaired ability of nuclei to polymerise RNA, reduction in template efficiency of chromatin, decrease in the activity of RNA polymerase and inhibition in histone phosphorylation. Protection of the spleen by lead shielding during whole-body exposure to X-rays largely eliminated the observed alterations in protein and nucleic acid synthetic machineries. This suggests that the radiation-induced inhibition in protein and RNA syntheses is mainly due to the direct action of radiation on the spleen itself.

Light-grown cells which contain fully developed chloroplasts were found to be more resistant to gamma-irradiation than dark-grown cells which are devoid of chloroplasts. The radio-resistance of dark-grown cells progressively increased during light-induced development of chloroplasts and, conversely, radio-resistance of light-grown cells decreased progressively with chloroplast de-development during growth in the dark. The presence of chloroplasts seemed to play a major role in the capacity of cells to recover from radiation damage, the efficiency of cellular recovery being correlatable with the degree of chloroplast development.

Exposure of rats to a single whole-body dose of X-rays led to an enhanced rate of RNA synthesis in the liver during 4 to 18 hours post-irradiation. This increase in RNA synthesis could not be obliterated by protecting the liver during irradiation. On the other hand, if adrenals were surgically removed prior to whole-body radiation exposure, no elevation either in liver RNA synthesis in vivo or liver chromatin template activity in vitro was apparent. Protection of only the head during irradiation could also prevent the stimulation in liver RNA synthesis in vivo. These results are suggestive of the involvement of neuro-endocrine reactions along the hypothalamus-pituitary-adrenal axes to be the underlying factors in whole-body radiation-evoked activation of the transcription machinery in the liver.

Erythrosine increased the yield of sporulation minus mutants of Bacillus subtilis excision repair-proficient strain 168 by approximately equal to 400% at a concentration of 1 mg/ml under ambient light conditions. This mutagenic response was dose-dependent (0-1 mg/ml). Significantly, the food dye did not mutate the corresponding repair-deficient B. subtilis, hcr-9 (exc-). A decrease in the mutagenicity of erythrosine to B. subtilis strain 168 was apparent on metabolism by rat liver S9 mixture or by rat caecal cell-free extracts. Erythrosine did not exhibit differential toxicity to B. subtilis excision repair-proficient (168) and -deficient (hcr-9) strains, although it was highly toxic to both strains. This indicated non-involvement of excision repair in the dye-mediated toxic reactions.

The formation of single-strand breaks by near-ultraviolet light at 313 nm and by aerobic gamma-rays was compared for skin fibroblast monolayer cultures from 4 normal donors (NF) and 8 patients with Bloom's syndrome (BS) by the alkaline elution method. In 6 of 8 BS strains, the number of breaks induced by near-ultraviolet light, 2.25 kJ/sq m, at 0 degrees was comparable to NF, while elevated breakage was observed in BS strains HG 369 and HG 916. Breakage frequencies were increased substantially in 6 of 8 BS strains relative to NF when the near-ultraviolet light exposure was at 37 degrees. BS strain GM 2520 represents an exception since normal breakage frequencies were induced both at 0 degrees and 37 degrees. Aerobic gamma-rays (75 R) induced comparable numbers of single-strand breaks in BS and NF strains at 0 degrees. The breakage frequencies were reduced an average of 17% in NF when the same dose was given at 30 degrees followed by 6 min incubation. Under the same conditions, the breakage frequencies were on the average reduced by 42% relative to 0 degrees in the BS strains, indicating that they possess normal or possibly slightly increased capacities for the rejoining of gamma-ray-induced breaks.

Erythrosine (diNa, tetraiodofluorescein) was nonmutagenic to the Ames/Salmonella typhimurium strains TA97a, TA98, TA100, TA102, and TA104, to a concentration of 2 mg/plate. No mutative intermediates were detected on metabolism by rat caecal cell-free extracts or rat liver S9 mixture; or on incubation with the comutagens, harman and norharman (+/- S9). Instead, an unexpected dose-dependent suppression in spontaneous reversion frequencies was observed (maximum approximately equal to 35% decrease). Erythrosine was antimutagenic to benzo[a]pyrene, but it did not decrease the mutagenicity of the other adduct-forming mutagen, 4-nitroquinoline N-oxide. The food dye was strongly antimutagenic to the bifunctional alkylating agent, mitomycin C, though it did not exhibit a similar effect on the mutagenicity of the corresponding monofunctional agent, methyl methanesulphonate. It partially depressed the mutagenic potentials of sodium azide. The antimutagenic effect of erythrosine on an intercalating agent, ethidium bromide, was discernible only at the highest dose (2 mg/plate). These results have been interpreted in terms of a genointeractive role of erythrosine. Erythrosine produced differential toxic effects in repair-deficient (TA97a, TA98, TA100) and repair-proficient (TA102, TA104) Salmonella tester strains; survival of the repair-deficient strains was found to be decreased. Photoinduced potentiation of erythrosine toxicity was observed, although light irradiation in the presence of erythrosine did not modify the reversion frequencies of the tester strains. The evidence strongly suggests that erythrosine, which exhibits nonmutagenicity in the Ames/Salmonella test, can interact with DNA repair enzymes and/or with DNA.

Administration of hydrocortisone to adrenalectomised rats elevated levels of both free and membrane-bound polysomes in liver and led to marked shifts in their sedimentation profiles in favour of heavier polysomes. The rate of nascent polypeptide formation was also found to be enhanced in response to the hormone but this effect was mainly confined to the site of free polysomes. These results suggest that besides its well-known potentiating action at the transcription level, the hormone elicits selective stimulatory response at the translation level.

Exposure of light-grown and dark-grown Euglena cells to gamma radiation causes single strand breaks in nuclear DNA as assessed by sedimentation analysis in alkaline sucrose density gradients. The number of radiation-induced single strand breaks in nuclear DNA of light-grown cells is found to be less than that in dark-grown cells. Post-irradiation incubation of both types of cells in 0 . 1 M phosphate buffer, pH 7 . 0 at 25 degrees C for 1 hour results in restitution of the strand breaks in DNA. Light-grown cells (cells with chloroplasts) are able to rejoin all the single strand breaks in DNA produced by gamma irradiation at D50 and D5 doses. On the other hand, dark-grown cells (cells devoid of chloroplasts) are unable to rejoin all the strand breaks caused by irradiation at either of the doses. The rate of DNA repair in dark-grown cells is also much slower than that in light-grown cells. Radiation-induced single strand breaks in DNA and their repair in nuclei from both types of cells is found to be similar to that observed in the spheroplasts. It is suggested that some factor(s) elaborated by chloroplasts may contribute towards the efficiency of nuclear DNA repair in Euglena cells.

Resdiual RNA synthesis in Escherichia coli cells and T(4)-infected E. coli cells exposed to ultraviolet radiation is insensitive to rifampicin. On the other hand, residual RNA synthesis in these cells after gamma irradiation is further inhibited by rifampicin. The specific effect exerted by ultraviolet irradiation on transcription seems to arise from alterations in the DNA structure rather than in bacterial RNA polymerase since RNA synthesis in ultraviolet-irradiated E. coli cells infected with unirradiated T(4) is sensitive to rifampicin.

Presence of insecticide-endosulfan, phosalone or permethrin in the growth medium caused concentration dependent inhibition in the vegetative growth of Chlamydomonas reinhardtii algal cells. The rate of inhibition produced by endosulfan was two-fold higher than that of phosalone or permethrin. Endosulfan affected the cell growth completely at 100 times less concentration as compared to that of phosalone or permethrin. Non-dividing cell populations encountered significant losses in cells during their exposure (2 h) to endosulfan and did not show further loss in the 72 h post-treatment period. The populations treated with phosalone exhibited losses of considerable magnitude in the post-treatment period. Permethrin treated non-diving cell populations did not lower the cell number, either after the treatment period (2 h) or the post-treatment period (72 h). However, these populations showed reduced levels of chlorophyll following the exposure of the insecticide and did not display recovery or further reduction in the levels in the post-treatment period. The chlorophyll levels of endosulfan or phosalone treated cell populations remained unaffected. The cells remaining intact after the treatment (2 h) of endosulfan or phosalone exhibited significant decreases in their post treatment vegetative growth abilities. The growth ability of such permethrin exposed cells was similar to that of untreated cells.

For 4--18 h following whole-body X-irradiation of rats, liver nuclei showed a progressive increase in the permeability to ribonucleoside triphosphates (as assessed in vitro using tritiated uridine triphosphate (UTP) and in the capacity to polymerise RNA in vitro (Mg++-containing and Mn++/(NH4)2SO4-containing assay systems).

Experiments on hybridization of [32P]HnRNA from hydrocortisone-treated rat liver with rat-liver DNA, in competition with unlabelled liver HnRNA from hormone-deficient and treated rats, revealed marked differences in their nucleotide sequence homologies. It was found that the hormone-induced new species of HnRNAs were transcribed both on middle-repetitive and non-repetitive sequences of DNA. However similar differences in the sequence homologies were not detectable in respect of mRNA derived from livers of hydrocortisone-administered and adrenalectomised rats. The results imply that the new species of HnRNA synthesized following hormone treatment may be confined to the nucleus.

Evidence has been presented to show that, like synthesis of RNA, syntheses of ribonucleotide precursors of RNA in the rat liver were stimulated for 6--18 h following whole-body X-irradiation (1000 R).

1. The heavy, light and fluffy mitochondrial fractions obtained by differential centrifugation were further characterized with respect to their protein synthesizing ability in vitro, their nucleic acid content, buoyant density of their DNA and ultrastructure. 2. The light mitochondrial fraction synthesized proteins in vitro at a rate 4-5 times as high as heavy and fluffy mitochondria. The incorporation ability of this fraction was also maximally affected by the thyroid status of the animal. The radioactivity in leucyl-tRNA of the light mitochondrial fraction was about 3-4 times as high as that of the other two fractions. 3. The heavy, light and fluffy mitochondrial fractions contained small but consistent amounts of RNA and DNA. Although the DNA content was the same in all mitochondria fractions, the light mitochondria contained relatively more RNA. The buoyant density of DNA from all the fractions was 1.701g/cm(3). 4. Electron microscopy revealed that the heavy mitochondria have a typical mitochondrial architecture, with densely packed cristae and a well developed double membrane. Light mitochondria were also surrounded by double membranes, but were smaller in size and contained less cristae. The fluffy fraction consisted of a mixture of well formed mitochondria and those in the process of degradation. 5. The significance of these findings in relation to mammalian mitochondrial genesis is discussed.