Heart rate variabilty changes during first week of acclimatization to 3500 m altitude in Indian military personnel.

Acute exposure to hypobaric hypoxia induces the changes in autonomic control of heart rate. Due to emergencies or war like conditions, rapid deployment of Indian military personnel into high altitude frequently occurs. Rapid deployment to high altitude soldiers are at risk of developing high altitude sickness. The present study was conducted to evaluate the acute exposure to high altitude hypobaric hypoxia (3500 m altitude) on the autonomic nervous control of heart rate in Indian military personnel during first week of acclimatization Indices of heart rate variability (viz; R-R interval, total power, low frequency, high frequency, ratio of low to high frequency) and pulse arterial oxygen saturation were measured at sea level and 3500m altitude. Power spectrum of heart rate variability was quantified by low frequency (LF: 0.04-0.15 Hz) and high frequency (HF: 0.15-0.5 Hz) widths. The ratio of LF to HF was also assessed as an index of the sympathovagal balance. Mean R-R interval decreased significantly on day 2 on induction to altitude which tended to increase on day 5. Total power (TP) decreased high altitude and tended to recover within a week. Both HF and LF power showed decrement at 3500m in comparison to sea level. The ratio of LF to HF (LF/HF) at 3500m was significantly higher at 3500m. SpO2 values decreased significantly (P < 0.05) at high altitude on day-2 which increased on day-5. We conclude that autonomic control of the heart rate measured by heart rate variability was altered on acute induction to 3500m which showed a significant decrease in parasympathetic tone and increase in sympathetic tone, then acclimatization seems to be characterized by progressive shift toward a higher parasympathetic tone.

Influence of hyperthermia on gamma-ray-induced mutation in V79 cells.

Asynchronously growing V79 cells were assayed for mutation induction following exposure to hyperthermia either immediately before or after being irradiated with 60Co gamma rays. Hyperthermia exposures consisted of either 43.5 degrees C for 30 min or 45 degrees C for 10 min. Each of these heat treatments resulted in a survival level of 42%. For all sequences of combined treatment with hyperthermia and radiation, cell killing by gamma rays was enhanced. Mutation induction by gamma rays was enhanced when heat preceded gamma irradiation, but no increase was observed when heat was given after gamma exposures. Treatment at 45 degrees C for 10 min gave a higher yield in mutants at all gamma doses studied compared to treatment at 43.5 degrees C for 30 min. When heat-treated cells were incubated for different periods before being exposed to gamma rays, thermal enhancement of radiation killing was lost after 24 h. In contrast, only 5-6 h incubation was needed for loss of mutation induction enhancement.

Supernatant medium from UV-irradiated cells influences the cytotoxicity and mutagenicity of V79 cells.

It is known that UV light induces the secretion of some proteins into the extracellular medium. We have carried out experiments to study how the supernatant medium from UV-irradiated cells affects the cytotoxicity and mutagenicity of V79 cells exposed to different damaging agents. So we exposed exponentially growing cells to 20 J/m2 of UV light and then harvested the supernatant medium after 22 h. This supernatant medium was then used to treat a fresh batch of cells for 2 h. After the treatment with this supernatant medium the cells were subsequently exposed to UV light, gamma-rays, hydrogen peroxide or MNNG. We found that exposure to this medium had a protective effect on the survival levels for UV light, gamma-rays and hydrogen peroxide while MNNG-induced killing remained unaffected. With UV light and gamma-rays we found that mutation induction at all doses was increased. Cycloheximide could inhibit this protection and the increase in mutation frequencies was also suppressed. The results indicated a protective role for the UV-induced factor(s). They were probably involved directly or they triggered repair process(es) that were related to oxidative stress.

Hyperthermia-induced modulation of killing and mutation by UV and N-methyl-N'-nitro-N-nitrosoguanidine in V79 cells.

Hyperthermic exposures of V79 cells did not affect the killing by UV light, whereas it enhanced MNNG-induced killing. Such hyperthermic exposure increased the mutation induction (resistance to 6-thioguanine) by both UV and MNNG. The timing of heat exposure, before or after the treatments, had no effect on the result in cases of cytotoxicity and mutagenesis.

Further studies on tetracycline-induced mutation in V79 Chinese hamster cells.

The interaction between ultraviolet light and tetracycline in producing cell killing and mutation has been studied in V79 Chinese hamster cells. It has been established that these agents act independently of each other. Cycloheximide altered the response to tetracycline in the fractionation experiment: when cycloheximide was not present, fractionation of TC treatment resulted in a higher mutation yield but no change in survival level; in the presence of cycloheximide, however, mutation was greatly reduced but survival increased. The results were taken to indicate that for tetracycline action to take place, de novo protein synthesis during tetracycline treatment was necessary. Caffeine had no influence on tetracycline-induced lethality or mutagenicity. This observation was considered to suggest that tetracycline did not affect cellular repair processes.

Evaluation of the genotoxicity of lac dye.

Red lac dye, a by-product of the shellac industry, has the potential for use in foods, drugs and cosmetics as a colouring agent. As part of a series of tests of the suitability of lac dye for this purpose an evaluation of its genotoxicity was carried out. Lac dye was non-mutagenic in Ames tests using five strains of Salmonella typhimurium with or without metabolic activation. No cytotoxicity or mutagenicity was observed in Chinese hamster lung (V79) cells exposed to lac dye in vitro. A clastogenic effect was observed in the bone-marrow cells of mice that had been treated with lac dye ip or orally.

Immediate treatment of frostbite using rapid rewarming in tea decoction followed by combined therapy of pentoxifylline, aspirin & vitamin C.

BACKGROUND & OBJECTIVES: Frostbite, the severest form of cold injury is a serious medical problem for our Armed Forces operating in the snow bound areas at high altitude. Effects of treatment by rapid rewarming in tea decoction followed by combined therapy of pentoxifylline, aspirin and vitamin C were evaluated in amelioration of tissue damage due to experimentally induced frostbite in rats. METHODS: Experiments were conducted in 2 groups (25 each) of albino rats (control i.e., untreated and experimental i.e., treated). Frostbite was produced experimentally in all the animals by exposing one of the hind limbs at -12 +/- 1 degree C with wind flow 25-30 lit/min for 30 min in a freezing-machine, with simultaneous recordings of rectal and ambient temperatures. The degree of tissue damage was assessed after 10 days. Following cold exposure, neither external thawing nor any medication was given to the animals of the control group; while the exposed limb of the experimental animals was rewarmed in tea decoction maintained at 37-39 degrees C for 30 min immediately after cold exposure, with simultaneous oral ingestion of warm tea decoction. These animals were also given pentoxifylline (40 mg/kg), aspirin (5 mg/kg) and vitamin C (50 mg/kg) twice daily orally for the next 7 days. RESULTS: In the control group, 68 per cent animals suffered from severe (56%) to very severe (12%) frostbite, while the remaining 32 per cent had moderate frostbite. No animals of this group could escape injury or suffered anything less than moderate frostbite; whereas 52 per cent of experimental animals escaped injury (no frostbite) and 32 and 16 per cent suffered only with primary and moderate degree of injury, respectively. None from this group suffered from severe or very severe frostbite. INTERPRETATION & CONCLUSION: It is evident from the study that this combined therapy resulted in significant improvement in the degree of tissue preservation and proved to be highly beneficial as an immediate treatment of frostbite in rats. The combined pharmacological properties of these drugs might have altered the haemorrheologic status of blood and produced curative beneficial effect in improving tissue survival. Clinical studies are required for confirmation of these beneficial effects in humans, which has already been taken up.

Effects of mountaineering training at high altitude (4,350 m) on physical work performance of women.

BACKGROUND: Little is known about work performance of women in hypobaric hypoxia. Moreover, whether native women of moderate altitude (2,000-2,100 m) differ from their lowland counterparts in their ability to adjust to hypobaric hypoxia is also not known. Hence, physiological alterations on work performance due to mountaineering training with altitude adaptation was evaluated in two groups of women and compared to the differences in the responses of the native women of moderate altitudes (Highlanders-HL) with those of the plains (Lowlanders-LL). METHODS: Pre-training tests were conducted at 2,100 m, then during sojourn to 4,350 m and re-tested again after return to 2,100 m. Physical work performance was assessed following standard step-test-exercise on a 30 cm stool with 24 cycles x min(-1) for 5 min. Heart rate, BP, ventilation, oxygen consumption and oxygen saturation were monitored at rest and during exercise followed by 5 min recovery in all three situations. RESULTS: During initial assessment, HL showed higher cardiovascular efficiency with faster recovery of exercise heart rate. Both groups showed significant improvement in physical performance due to mountaineering training at high altitude (HA). The difference in performance between two groups narrowed down at 4,350 m and further reduced during re-test with maintenance of initial superiority of the HL. CONCLUSIONS: a) Native women of moderate altitude (HL) are more fit compared with their plains counterparts (LL); b) All women achieved marked improvement in cardiovascular and respiratory efficiency as well as the step-test score due to intense mountaineering training at HA, and the rate of improvement in physical performance was higher in LL; c) Further, induction by trekking under progressive hypoxia coupled with rigorous mountaineering activity at HA merits in understanding better acclimatization and improved physical performance.

Chronic low dose exposure to hydrogen peroxide changes sensitivity of V79 cells to different damaging agents.

Chinese hamster V79 cells were repeatedly exposed to a low dose of hydrogen peroxide (H2O2) over several weeks and then exposed to H2O2, cisplatin or ultraviolet (UV) light. Cell killing was examined by colony formation, following these treatments. It was seen that cells conditioned by multiple low doses of H2O2 showed resistance to killing in case of H2O2 and cisplatin but the sensitivity to UV light was same as the control cells. Apoptosis was also determined in these cells after the same treatments. UV light failed to induce apoptosis in both conditioned and in control cells, but in case of cells treated with H2O2 and with cisplatin, there was less apoptosis in the conditioned cells compared to the control cells. From our observation we can say that the enhanced survival of cells after treatment with H2O2 or cisplatin could be due to inhibition of apoptosis.

Oxygen saturation response to exercise VO2 at 2100 m and 4350 m in women mountaineering trainees.

Human work performances decreases at high altitude (HA). This decrement does not appear to be similar for every individual, may be due to variety of factors like elevation, mode of induction, work intensity, physical condition and specificity of the subjects. The purpose of the study was to evaluate the effects of alteration in responses of oxygen saturation (SaO2) and oxygen consumption (VO2) to a standard exercise in women mountaineering trainees under hypobaric hypoxia. Experiments were conducted in 2 groups (10 each) of females and compared the difference in responses of native women of moderate altitude with those of the plains/low altitude. A standard exercise test (Modified Harvard Step-Test for women) was performed on a 30 cm stool with 24 cycles/min for 5 min, initially at 2100 m and then at 4350 m. The exercise VO2 values for plains dwelling women achieved apparently VO2max level at both altitude locations with significant reduction in SaO2 during standard exercise. Exercise VO2 values decreased on exposure to 4350 m with further reduction in SaO2. Whereas with same work intensity, under same situation the exercise VO2 values of the moderate altitude women did not appear to have reached VO2max. They also maintained comparatively higher level of SaO2. It may be concluded that hypoxic exposure along with mountaineering training, the moderate altitude women maintained a higher level of SaO2 during standard exercise at both altitude locations, compared to low altitude women who might have lost a compensatory reserve to defend the hypoxic stress to exercise. Thus, moderate altitude women are proved to be better fit for hypoxic tolerance/HA performance.

Inhibition of calcium signaling in ultraviolet-irradiated fibroblasts: role of tyrosine phosphorylation and protein kinase C.

Our aim was to study whether ultraviolet radiation produced any alterations in the subsequent signaling response of V79 fibroblasts to mitogenic stimulus. In ultraviolet C (UVC)-irradiated V79 fibroblasts, increase in cytosolic calcium in response to thrombin was nearly abolished in the presence of 3 mM external Ca(2+). UVC-treated V79 cells showed a greatly enhanced permeability to Ca(2+) which was reversed by pretreatment with genistein, a tyrosine kinase inhibitor. Genistein also alleviated the inhibition of thrombin response caused by UVC. In UVC-treated cells, significant activation of protein kinase C (PKC) occurred only on exposure to 3 mM external calcium and PKC inhibitors (H-7 or staurosporine) reversed UVC-induced adverse effects on the thrombin response. Therefore, it is likely that protein tyrosine phosphorylation by UVC may play a role in the subsequent inhibition of thrombin response in V79 cells through increased calcium influx and activation of PKC.

Repair of heat injury in thymine auxotrophs of Escherichia coli.

Heat-induced damage in four strains of bacteria has been studied. It has been shown that the damage is repairable on incubation at 37 degrees C in a medium not supporting growth. The extent of repair is dependent upon the strain of bacterium chosen. Recovery from the damage is prominent in the case of filament-forming bacteria, and in non-filament formers, recovery is not totally absent.

Thrombin releases calcium from internal stores of ultraviolet C-treated V79 fibroblasts independent of phosphatidylinositol bisphosphate hydrolysis: role of oxidative stress.

V79 fibroblasts were treated with ultraviolet (UV) C radiation alone as well as in conjunction with chronic oxidative stress. The effects of these treatments on calcium signaling were observed at 30 min post-irradiation. In the absence of extracellular calcium, thrombin released calcium from internal stores of UVC-irradiated V79 fibroblasts even after exposure to neomycin. In neomycin-treated control and chronic oxidative stress cells, no calcium release by thrombin was observed after chelation of external calcium. Calcium release by thrombin from internal stores of UV-irradiated and neomycin-treated cells was completely abolished by pretreatment with N-acetyl cysteine and dexamethasone. Cellular total soluble thiol content which is a good indicator of cellular reduced glutathione (GSH) level was significantly elevated 30 min after ultraviolet radiation, indicating an adaptive response after oxidative stress. Chronic oxidative stress alone resulted in a much smaller increase in GSH but chronic oxidative stress in conjunction with UVC produced a very prominent elevation in GSH levels. Our data suggest that thrombin can cause calcium release from internal stores of ultraviolet-irradiated fibroblasts which is independent of phosphatidylinositol bisphosphate hydrolysis and is directly related to the level of oxidative stress. Involvement of phospholipase A2 and a role for its products as possible mediators of calcium release from intracellular stores, is strongly indicated.

UV non-mutable mutant from V-79 Chinese hamster cells.

To get an idea about the response of a living system, exposed to gradually increasing doses of a mutagen for several generations, a population of V-79 Chinese hamster cells was exposed repeatedly to gradually increasing doses of UV radiation. Each dose was followed by a variable period of growth for at least ten generations. After treatment the cells were not mutable by UV radiation, though MNNG was capable of producing mutations with the same efficiency as in the untreated cells. In terms of viability, the treated cells behaved exactly as the untreated ones for both UV and MNNG. The observed behaviour of the treated cells was found to be stable for during the 50 passages studied.

The role of free radicals in cold injuries.

Cold injury is a tissue trauma produced by exposure to freezing temperatures and even brief exposure to a severely cold and windy environment. Rewarming of frozen tissue is associated with blood reperfusion and the simultaneous generation of free oxygen radicals. In this review is discussed the current understanding of the mechanism of action of free oxygen radicals as related to cold injury during rewarming. Decreased energy stores during ischaemia lead to the accumulation of adenine nucleotides and liberation of free fatty acids due to the breakdown of lipid membranes. On rewarming, free fatty acids are metabolized via cyclo-oxygenase and adenine nucleotides are metabolized via the xanthine oxidase pathway. These may be the source of free oxygen radicals. Leukocytes may also play a major role in the pathogenesis of cold injury. Oxygen radical scavengers, such as superoxide dismutase and catalase, may help to reduce the cold induced injury but their action is limited due to the inability readily to cross the plasma membrane. Lipid soluble antioxidants are likely to be more effective scavengers because of their presence in membranes where peroxidative reactions can be arrested.