Surfactant protein A suppresses reactive nitrogen intermediates by alveolar macrophages in response to Mycobacterium tuberculosis.
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Mycobacterium tuberculosis attaches to, enters, and replicates within alveolar macrophages (AMs). Our previous studies suggest that surfactant protein A (SP-A) can act as a ligand in the attachment of M. tuberculosis to AMs. Reactive nitrogen intermediates (RNIs) play a significant role in the killing of mycobacteria. We have demonstrated that RNI levels generated by AMs were significantly increased when interferon-gamma-primed AMs were incubated with M. tuberculosis. However, the RNI levels were significantly suppressed in the presence of SP-A (10 microg/ml). The specificity of SP-A's effect was demonstrated by the use of F(ab')2 fragments of anti-SP-A monoclonal antibodies and by the use of mannosyl-BSA, which blocked the suppression of RNI levels by SP-A. Furthermore, incubation of deglycosylated SP-A with M. tuberculosis failed to suppress RNI by AMs, suggesting that the oligosaccharide component of SP-A, which binds to M. tuberculosis, is necessary for this effect. These results show that SP-A-mediated binding of M. tuberculosis to AMs significantly decreased RNI levels, suggesting that this may be one mechanism by which M. tuberculosis diminishes the cytotoxic response of activated AMs. (+info)
Effects of nucleoside analog incorporation on DNA binding to the DNA binding domain of the GATA-1 erythroid transcription factor.
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We investigate here the effects of the incorporation of the nucleoside analogs araC (1-beta-D-arabinofuranosylcytosine) and ganciclovir (9-[(1,3-dihydroxy-2-propoxy)methyl] guanine) into the DNA binding recognition sequence for the GATA-1 erythroid transcription factor. A 10-fold decrease in binding affinity was observed for the ganciclovir-substituted DNA complex in comparison to an unmodified DNA of the same sequence composition. AraC substitution did not result in any changes in binding affinity. 1H-15N HSQC and NOESY NMR experiments revealed a number of chemical shift changes in both DNA and protein in the ganciclovir-modified DNA-protein complex when compared to the unmodified DNA-protein complex. These changes in chemical shift and binding affinity suggest a change in the binding mode of the complex when ganciclovir is incorporated into the GATA DNA binding site. (+info)
Effects of the Chinese traditional medicine mao-bushi-saishin-to on therapeutic efficacy of a new benzoxazinorifamycin, KRM-1648, against Mycobacterium avium infection in mice.
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The Chinese traditional medicine mao-bushi-saishin-to (MBST), which has anti-inflammatory effects and has been used to treat the common cold and nasal allergy in Japan, was examined for its effects on the therapeutic activity of a new benzoxazinorifamycin, KRM-1648 (KRM), against Mycobacterium avium complex (MAC) infection in mice. In addition, we examined the effects of MBST on the anti-MAC activity of murine peritoneal macrophages (M phi s). First, MBST significantly increased the anti-MAC therapeutic activity of KRM when given to mice in combination with KRM, although MBST alone did not exhibit such effects. Second, MBST treatment of M phi s significantly enhanced the KRM-mediated killing of MAC bacteria residing in M phi s, although MBST alone did not potentiate the M phi anti-MAC activity. MBST-treated M phi s showed decreased levels of reactive nitrogen intermediate (RNI) release, suggesting that RNIs are not decisive in the expression of the anti-MAC activity of such M phi populations. MBST partially blocked the interleukin-10 (IL-10) production of MAC-infected M phi s without affecting their transforming growth factor beta (TGF-beta)-producing activity. Reverse transcription-PCR analysis of the lung tissues of MAC-infected mice at weeks 4 and 8 after infection revealed a marked increase in the levels of tumor necrosis factor alpha, gamma interferon (IFN-gamma), IL-10, and TGF-beta mRNAs. KRM treatment of infected mice tended to decrease the levels of the test cytokine mRNAs, except that it increased TGF-beta mRNA expression at week 4. MBST treatment did not affect the levels of any cytokine mRNAs at week 8, while it down-regulated cytokine mRNA expression at week 4. At week 8, treatment of mice with a combination of KRM and MBST caused a marked decrease in the levels of the test cytokines mRNAs, especially IL-10 and IFN-gamma mRNAs, although such effects were obscure at week 4. These findings suggest that down-regulation of the expression of IL-10 and TGF-beta is related to the combined therapeutic effects of KRM and MBST against MAC infection. (+info)
Steady-state nitrogen isotope effects of N2 and N2O production in Paracoccus denitrificans.
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Nitrogen stable-isotope compositions (delta15N) can help track denitrification and N2O production in the environment, as can knowledge of the isotopic discrimination, or isotope effect, inherent to denitrification. However, the isotope effects associated with denitrification as a function of dissolved-oxygen concentration and their influence on the isotopic composition of N2O are not known. We developed a simple steady-state reactor to allow the measurement of denitrification isotope effects in Paracoccus denitrificans. With [dO2] between 0 and 1.2 microM, the N stable-isotope effects of NO3- and N2O reduction were constant at 28.6 per thousand +/- 1.9 per thousand and 12.9 per thousand +/- 2.6 per thousand, respectively (mean +/- standard error, n = 5). This estimate of the isotope effect of N2O reduction is the first in an axenic denitrifying culture and places the delta15N of denitrification-produced N2O midway between those of the nitrogenous oxide substrates and the product N2 in steady-state systems. Application of both isotope effects to N2O cycling studies is discussed. (+info)
Experiment of nitrox saturation diving with trimix excursion.
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Depth limitations to diving operation with air as the breathing gas are well known: air density, oxygen toxicity, nitrogen narcosis and requirement for decompression. The main objectives of our experiment were to assess the decompression, counterdiffusion and performance aspect of helium-nitrogen-oxygen excursions from nitrox saturation. The experiment was carried out in a wet diving stimulator with "igloo" attached to a 2-lock living chamber. Four subjects of two teams of 2 divers were saturated at 25 msw simulated depth in a nitrogen oxygen chamber environment for 8 days, during which period they performed 32 divers-excursions to 60 or 80 msw pressure. Excursion gas mix was trimix of 14.6% oxygen, 50% helium and 35.4% nitrogen, which gave a bottom oxygen partial pressure of 1.0 bars at 60 msw and 1.3 at 80 msw. Excursions were for 70 min at 60 msw with three 10-min work periods and 40 min at 80 msw with two 10-min work periods. Work was on a bicycle ergometer at a moderate level. We calculated the excursion decompression with M-Values based on methods of Hamilton (Hamilton et al., 1990). Staged decompression took 70 min for the 60 msw excursion and 98 min for 80 msw, with stops beginning at 34 or 43 msw respectively. After the second dive day bubbles were heard mainly in one diver but in three divers overall, to Spencer Grade III some times. No symptoms were reported. Saturation decompression using the Repex procedures began at 40 msw and was uneventful: Grade II and sometimes III bubbles persisted in 2 of the four divers until 24 hr after surfacing. We conclude that excursions with mixture rich in helium can be performed effectively to as deep as 80 msw using these procedures. (+info)
Nitrogen retention by lambs fed oscillating dietary protein concentrations.
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Nitrogen excreted by beef cattle can be retained in manure or lost by volatilization to the atmosphere or by runoff and percolation into surface or ground water. Increasing the retention of dietary N should decrease environmental losses. To this end, the effects of oscillating concentrations of dietary CP on nutrient retention were determined using lambs fed a 90% concentrate diet. Ten St. Croix lambs (average BW = 27 kg) were used in two 5x5 Latin square experiments. Dietary treatments were as follows: 1) 10% CP, 2) 12.5% CP, 3) 15% CP, 4) 10% and 15% CP diets oscillated at 24-h intervals, and 5) 10% and 15% CP diets oscillated at 48-h intervals. Supplemental N was provided by cottonseed meal in Trial 1 and by a 50:50 (N basis) blend of cottonseed meal and urea in Trial 2. Each period of the Latin square lasted 35 d, with excreta collection the final 8 d. Nitrogen retention increased linearly (P<.01) with increasing N intake in both trials (.77, 1.33, and 1.89 g/d for 10, 12.5, and 15% CP, respectively, in Trial 1; .94, 1.78, and 2.19 g/d for 10, 12.5, and 15% CP, respectively, in Trial 2). Compared with continuously feeding the 12.5% CP diet, oscillating the 10 and 15% CP diets on a 24-h basis did not affect N retention (P>.10) in either trial (1.62 and 1.56 g/d for Trials 1 and 2, respectively). Oscillating dietary CP at 48-h intervals did not affect N retention in Trial 2 (1.82 g/d) but increased (P<.05) N retention by 38% in Trial 1 (1.87 g/d). Phosphorus, K, and Na retention and excretion were not affected by dietary treatments in Trial 1. In Trial 2, P retention increased (linear, P<.05) with increasing dietary CP and was greater (P<.05) in lambs on the 48-h oscillation treatment than in lambs fed the 12.5% CP diet. These results suggest that oscillating the dietary CP concentrations might potentially increase the utilization of N by ruminants fed high-concentrate diets. (+info)
A comparative chemical and histochemical study of the chondrodystrophoid and nonchondrodystrophoid canine intervertebral disc.
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The chemical composition of the intervertebral disc of 9-month-old chondrodystrophoid and nonchondrodystrophoid dogs was studied for collagen, noncollagenous protein and glycosaminoglycan. Content of these substances differed significantly between breeds. The differences were most marked in the nucleus pulposus; the noncollagenous protein content of the nonchondrodystrophoid breed was higher than in that of the chondrodystrophoid dogs. The total nitrogen value of the nonchondrodystrophoid nuclei pulposi was less than that of the corresponding chondrodystrophoid discs mainly because of the high collagen content of the latter discs. Histochemically, it was found that the nuclei pulposi of the nonchondrodystrophoid breed contains larger amounts of glycosaminoglycan than in the discs of the chondrodystrophoid breeds. (+info)
Kinetic impairment of nitrogen and muscle glutamine metabolisms in old glucocorticoid-treated rats.
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Aged rats are more sensitive to injury, possibly through an impairment of nitrogen and glutamine (Gln) metabolisms mediated by glucocorticoids. We studied the metabolic kinetic response of adult and old rats during glucocorticoid treatment. The male Sprague-Dawley rats were 24 or 3 mo old. Both adult and old rats were divided into 7 groups. Groups labeled G3, G5, and G7 received, by intraperitoneal injection, 1.50 mg/kg of dexamethasone (Dex) for 3, 5, and 7 days, respectively. Groups labeled G3PF, G5PF, and G7PF were pair fed to the G3, G5, or G7 groups and were injected with an isovolumic solution of NaCl. One control group comprised healthy rats fed ad libitum. The response to aggression induced specifically by Dex (i.e., allowing for variations in pair-fed controls) appeared later in the aged rats (decrease in nitrogen balance from day 1 in adults but only from day 4 in old rats). The adult rats rapidly adapted to Dex treatment, whereas the catabolic state worsened until the end of treatment in the old rats. Gln homeostasis was not maintained in the aged rats; despite an early increase in muscular Gln synthetase activity, the Gln pool was depleted. These results suggest a kinetic impairment of both nitrogen and muscle Gln metabolisms in response to Dex with aging. (+info)