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Nitric monoxide (NO) exerts a great variety of physiological functions. L-Arginine supplies amino groups which are transformed to NO in various NO-synthase-active isoenzyme complexes. NO-synthesis is stimulated under various conditions increasing the tissue of stable NO-metabolites. The major oxidation product found is nitrite. Elevated nitrite levels were reported to exist in a variety of diseases including HIV, reperfusion injury and hypovolemic shock. Denitrifying bacteria such as Paracoccus denitrificans have a membrane bound set of cytochromes (cyt cd1, cyt bc) which were shown to be involved in nitrite reduction activities. Mammalian mitochondria have similar cytochromes which form part of the respiratory chain. Like in bacteria quinols are used as reductants of these types of cytochromes. The observation of one-e- divergence from this redox-couple to external dioxygen made us to study whether this site of the respiratory chain may also recycle nitrite back to its bioactive form NO. Thus, the aim of the present study was therefore to confirm the existence of a reductive pathway which reestablishes the existence of the bioregulator NO from its main metabolite NO2-. Our results show that respiring mitochondria readily reduce added nitrite to NO which was made visible by nitrosylation of deoxyhemoglobin. The adduct gives characteristic triplet-ESR-signals. Using inhibitors of the respiratory chain for chemical sequestration of respiratory segments we were able to identify the site where nitrite is reduced. The results confirm the ubiquinone/cyt bc1 couple as the reductant site where nitrite is recycled. The high affinity of NO to the heme-iron of cytochrome oxidase will result in an impairment of mitochondrial energy-production. "Nitrite tolerance" of angina pectoris patients using NO-donors may be explained in that way.
The aim of the present study was to establish the effects of platelet-activating factor (PAF) on selected movement parameters, plasmalemma integrity, capacitation process and acrosome reaction in cryopreserved boar spermatozoa. A positive effect of PAF addition to cryopreserved semen on sperm motility was demonstrated, particularly with the application of phospholipid concentration of 1 x 10-6M-1 x 10-5M. A moderate induction of plasmalemma damage of cryopreserved spermatozoa was observed when PAF was used at a low concentration (1 x 10-8M-1 x 10-7M). The rate at which PAF induced the process of capacitation was inversely proportional to its concentration in the sample (the highest for the concentration of 1 x 10-8M, and the lowest at 1 x 10-5M). In turn, the strongest induction of acrosome reaction of spermatozoa was observed in samples with the addition of PAF at a concentration of 1 x 10-7M. The results obtained suggest that the application of PAF supplement to post-thawed boar semen can be used as a laboratory test of the ability of spermatozoa to induce the acrosome reaction.
The localisation of carletinin in the midbrains of 10 sexually mature chinchilla males was examined by carrying out the peroxidase-antiperoxidase immunocytochemical reaction using specific monoclonal antibody against calretinin. Intensive immunostaining for the protein was observed in the majority of fusiform, pyramidal, and stellate neurons of dorsal raphe nucleus. A similar calretinin distribution in neurons of this region to those observed in primates and rodents was demonstrated.
Melatonin plays several important physiological functions in mammals, such as immune enhancement and regulation of dark-light signal transduction. Melatonin is also known to be an endogenous free radical scavenger and an efficient antioxidant. It detoxifies a variety of free radicals and reactive oxygen intermediates, including the hydroxyl radical, singlet oxygen and nitric oxide. These radicals participate in many diseases, for example diabetes. This study determined the effect of melatonin on the antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), and the level of glutathione (GSH) in human diabetic (C2 line) skin fibroblasts. Confluent monolayers of control (S2 line) and diabetic (C2 line) skin fibroblasts were incubated with different concentrations of melatonin: 10, 50, 100 and 1000 μmol/l at 37°C for 24 h. Next, the GSH level and SOD, CAT and GPx activities were measured colorimetrically. The activities of the antioxidant enzymes and the GSH level were lower in diabetic skin fibroblasts than in the control S2 line. Concentrations of melatonin of 100 and 1000 μmol/l caused a significant increase in the enzymes' activities and GSH level.
The metal ion coordination abilities of reduced and oxidized glutathione are reviewed. Reduced glutathione (GSH) is a very versatile ligand, forming stable complexes with both hard and soft metal ions. Several general binding modes of GSH are described. Soft metal ions coordinate exclusively or primarily through thiol sulfur. Hard ones prefer the amino acid-like moiety of the glutamic acid residue. Several transition metal ions can additionally coordinate to the peptide nitrogen of the γ-Glu-Cys bond. Oxidized glutathione lacks the thiol function. Nevertheless, it proves to be a surprisingly efficient ligand for a range of metal ions, coordinating them primarily through the donors of the glutamic acid residue.
Most clinical variables in animals follow biological rhythms which have a mathematical function defined by cosmic-climatic rhythms. Chronophysiology, chronopathology, chronopharmacology and chronotherapy have common elements but are frequently studied in isolation, thus making it difficult for a global understanding of clinical chronobiology as a unitary and well-defined discipline. The physiological effects of a drug depend not only on its molecular structure but also on the time-pattern of its administration. One of the main reasons for the importance of temporal patterns in drug activity is biological rhythm, and in particular that of the circadian period. These rhythms affect most physiological functions as well as drug metabolism, clearance, and dynamic processes that may alter drug availability and target cell responsiveness in relation to biological time. Chronotherapy studys the optimal level of drug effects and/or the minimizing of its toxicity by timing medications in relation to biological rhythms. This review focuses on medical chronobiology, which is much more technical since it only studies those aspects of clinical chronobiology having a health-care impact on daily practice.
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