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1.
Dynamics of hemoglobin, ceruloplasmin concentration, the changes of chemiluminescence in blood plasma and kinetics of rat erythrocyte heat denaturation during consequent exposition of high altitude hypoxia and hyperbaric oxygenation have been studied. Severe hypoxia causes the decrease of extraerythrocyte hemoglobin and oxidase activity of ceruloplasmin. Reoxygenation results in significant increase of blood plasma chemiluminescence with simultaneous increase of extraerythrocyte hemoglobin level and with modification of surface structure of the erythrocyte membranes. Possible pathways of activation of oxygen-dependent of free-radical processes during reoxygenation are discussed.  相似文献   

2.
A thin-layer gas-solution microcalorimeter has been developed to study the binding reactions of gaseous ligands with ligand binding macromolecules. We have measured the enthalpy of binding oxygen and carbon monoxide to horse myoglobin, human hemoglobin A0 and sperm whale myoglobin in phosphate buffer at pH 7.6, with the enzyme reducing system of Hayashi. Reactions of human hemoglobin were also done under various buffer conditions in order to elucidate the Bohr effect. These binding reactions were found not to exhibit a detectable enthalpy change over the temperature range of 10 degrees C to 25 degrees C. The enzyme reducing system was shown to react with oxygen in a manner that releases a substantial amount of heat. This problem was corrected by using a minimum amount and by placing the buffer and enzyme system in the reference cell effectively cancelling the oxygen enzyme reaction heat as well as the heat of gas dissolution. It was also demonstrated that glucose-6-phosphate, one of the reducing system components, in 50 mM concentrations can influence the heat of binding oxygen and carbon monoxide to hemoglobin. This effect was shown to be absent in the myoglobins and also with hemoglobin at glucose-6-phosphate concentrations less than 5 mM.  相似文献   

3.
Data in the literature based on the technique of graded osomotic hemolysis have been re-evaluated. Differences were previously found in the glucose-6-phosphate dehydrogenase/hemoglobin ratio and in the heat stability of the enzyme in hemolysates of 'old' and 'young' cells. These differences were believed to be due to the aging of the enzyme. As the erythrocyte membrane acts as a molecular sieve under hypotonic conditions [cf. Cseke, E., Váradi, A., Szabolcsi, G., and Biszku, E. (1978) FEBS Lett. 96, 15--18], the hemolysate obtained when a fraction is lysed does not properly represent the content of the lysed cells. As hemoglobin is lost from cells which are not yet lysed, the enzyme/hemoglobin ratio is underestimated in 'old' cells and overestimated in 'young' cells. It is further shown that the observed differences in the heat stability of glucose-6-phosphate dehydrogenase in the fractions obtained by graded hemolysis are due to the presence of different concentrations of endogeneous NADP. Therefore the published data obtained by graded osmotic hemolysis do not prove the assumption that the enzyme is aging during the lifetime of the erythrocyte.  相似文献   

4.
In an effort to assess the effects of environmental heat stress on muscle metabolism during exercise, 6 men performed work in the heat (Tdb = 44 degrees C, RH = 15%) and cold (Tdb = 9 degrees C, RH = 55%). Exercise consisted of three 15-min cycling bouts at 70 to 85% VO2max, with 10-min rest between each. Muscle biopsies obtained from the vastus lateralis before and after each work bout were analyzed for glycogen and triglyceride content. Venous blood samples drawn before and after exercise were assayed for lactate, glucose, free fatty acids, hemoglobin, and hematocrit. Oxygen uptake, heart rates and rectal temperatures were all significantly higher during exercise in the heat. Blood lactate concentration was roughly twice as great during the heat experiments as that measured in the 9 degrees C environment. Muscle glycogen utilization per 60 min was significantly greater in the heat ( - 74 m moles/kg-wet muscle) as compared to the cold exercise (- 42 m moles/kg-wet muscle). On the average, muscle triglyceride declined 23% during exercise in the cold and 11% in the heat. The findings of an enhanced glycolysis during exercise in the heat is compatible with earlier studies which demonstrate a decreased availability of oxygen due to a reduction in muscle blood flow.  相似文献   

5.
Six male subjects exercised for 50 min at 25% (light exercise) and 55% (moderate exercise) of their estimated aerobic capacities in environments of 42 degrees C db, 35 degrees C wb and 30 degrees C db, 24 degrees C wb, respectively. Alterations in the hematocrit, hemoglobin, and plasma protein concentrations, and in the activity of an injected aliquot of isotopically labeled albumin were each used to calculate the percentage change in plasma volume occurring during exercise and recovery. Changes in each measure were consistent with a reduction in plasma volume during exercise and a return to preexercise levels during recovery. There was no significant difference between the measures when exercising in the heat, but during the more severe exercise in the cooler environment disproportional changes in protein, hematocrit, and hemoglobin were observed. Disproportional changes were also seen during the recovery phase, when the hematocrit and hemoglobin concentration indicated a more rapid return of the plasma volume to preexercise levels than did either the plasma protein concentration or albumin activity. During moderate exercise and recovery there was a 1% decrease in red cell volume. It is concluded that exercise accelerates the rate of protein movement from extravascular compartments to the intravascular compartment, leading to elevated plasma protein levels during recovery which favor the return of water to the intravascular space. Hemoglobin concentration is considered to be the most reliable measure of plasma volume change during exercise.  相似文献   

6.
H Wajcman  G Gacon  D Labie  R D Koler  R T Jones 《Biochemistry》1975,14(22):5017-5020
Hemoglobin Casper (beta106Leu replaced by Pro) can be separated from hemoglobin (Hb) A by isoelectric focusing on polyacrylamide gel. This abnormal hemoglobin was estimated to be 30% of teh total by both isoelectric focusing and heat lability kinetics. Its oxygen equilibrium curves indicate a high oxygen affinity, low degree of subunit interaction, and a decreased Bohr effect. Mixtures of Hb Casper and Hb A appear to bind oxygen as if no hybrid molecules exist.  相似文献   

7.
Guan SM  Nagata H  Shizukuishi S  Wu JZ 《Anaerobe》2006,12(5-6):279-282
In this study, the ability of Prevotella intermedia, an obligate anaerobic rod, to degrade human hemoglobin was determined by SDS-PAGE and the degradation was quantified by scanning densitometry. Both bacterial cells and culture supernatants degraded hemoglobin. The hemoglobin degradation by P. intermedia was time-dependent, heat sensitive, pH related and was not influenced by iron restriction. Inhibition studies demonstrated that a cysteine protease might be involved in hemoglobin degradation and this protease might require metal ions for its activity and it might be thiol-requiring and trypsin-inducible. The results indicate that P. intermedia is capable to release heme from hemoglobin, hence provide a source of iron for its proliferation.  相似文献   

8.
Previous results [(1988) Arct. Med. Res. 47, 83-88] have shown that hemoglobin from reindeer is characterized by a low overall heat of oxygenation. This particular aspect has been investigated further in a series of precise oxygen equilibrium experiments. The results obtained show a peculiar dependence of the temperature effect on the fractional saturation of hemoglobin with oxygen, which could be regarded as a very interesting case of molecular adaptation to extreme environmental conditions.  相似文献   

9.
This study investigates the effects of a short-term aerobic training program in a hot environment on thermoregulation, blood parameters, sweat secretion and composition in tropic-dwellers who have been exposed to passive heat. Sixteen healthy Malaysian-Malay male volunteers underwent heat acclimation (HA) by exercising on a bicycle ergometer at 60% of VO2max for 60 min each day in a hot environment (Ta: 31.1+/-0.1 degrees C, rh: 70.0+/-4.4%) for 14 days. All parameters mentioned above were recorded on Day 1 and at the end of HA (Day 16). On these two days, subjects rested for 10 min, then cycled at 60% of VO2max for 60 min and rested again for 20 min (recovery) in an improvised heat chamber. Rectal temperature (Tre), mean skin temperature (Tsk) heart rate (HR), ratings of perceived exertion (RPE), thermal sensation (TS), local sweat rate and percent dehydration were recorded during the test. Sweat concentration was analysed for sodium [Na+]sweat and potassium. Blood samples were analysed for biochemical changes, electrolytes and hematologic indices. Urine samples were collected before and after each test and analysed for electrolytes.After the period of acclimation the percent dehydration during exercise significantly increased from 1.77+/-0.09% (Day 1) to 2.14+/-0.07% (Day 16). Resting levels of hemoglobin, hematocrit and red blood cells decreased significantly while [Na+]sweat increased significantly. For Tre and Tsk there were no differences at rest. Tre, HR, RPE, TS, plasma lactate concentration, hemoglobin and hematocrit at the 40th min of exercise were significantly lower after the period of acclimation but mean corpuscular hemoglobin and serum osmolality were significantly higher while no difference was seen in [Na+]sweat and Tsk. It can be concluded that tropic-dwelling subjects, although exposed to prolonged passive heat exposure, were not fully heat acclimatized. To achieve further HA, they should gradually expose themselves to exercise-heat stress in a hot environment.  相似文献   

10.
A "fetal hemoglobin' has been reported to exist during mouse gestation, Investigations using CMC chromatography, starch gel electrophoresis or isoelectric focusing have shown a hemoglobin band from fetal tissues, and blood was obtained which was different from the adult hemoglobin and designated a "fetal hemoglobin'. In the current study isoelectric focusing was used to study the hemoglobins existing in the tissues and blood during fetal and neonatal development and the results suggest there is no "fetal hemoglobin' present during gestation. It appears that the hemoglobin designated as "fetal' in our laboratory was a methemoglobin formed by an incomplete reaction of KCN with the hemoglobin. The additional hemoglobin bands which were obtained from fetal liver or neonatal spleen tissues appeared to be a modified adult hemoglobin.  相似文献   

11.
The oxygen-binding characteristics and the multiplicity of the stripped hemoglobiin from active lungfish Protopterus amphibius, are the same as in specimens that have been estivating for about 30 months, showing that alteration in the hemoglobin molecules is not involved in the earlier reported increase in oxygen affinity of whole blood during estivation (Johansen et al., '76). At pH 7.0 and 26 degrees C the hemolysates show a high oxygen affinity (P50 = 3.1 Torr), a Bohr factor (delta log P50/delta pH) of - 0.33, and a cooperativity coefficient (n) of 1.7. Between 15 and 26 degrees C, the apparent heat of oxygenation (delta H) is - 8.6 Kcal-mole-1 at pH 7.0, corresponding with data for other fish. A low sensitivity of oxygen affinity to urea appears to be adaptive to the high urea concentrations in estivating lungfish. The salt sensitivity is, however, similar to human hemoglobin. The hemoglobin consists of two major (electrophoretically anodal) components, which differ slightly in oxygen affinity but are both sensitive to pH and nucleoside triphosphates (NTP). Guanosine triphosphate (GTP), the major erythrocytic organic phosphate, however, depresses the oxygen affinity of the composite and separated hemoglobins more effectively than ATP suggesting that GTP is the primary modulator of oxygen affinity. Comparative measurements reveal only one major hemoglobin component in P. annectens which has a markedly lower oxygen affinity and phosphate sensitivity than P. amphibius hemoglobins and thus seems less pliable to phosphate-mediated variation in oxygen affinity. The data are discussed in relation to the hemoglobin systems of other fish.  相似文献   

12.
The investigation of two thermal properties of red cells throws some light on whether sickling is a process involving the crystallization of a relatively insoluble hemoglobin. These properties are the specific heat and the heat of compression, both of which would be expected to become numerically less if the hemoglobin of the red cell were to crystallize. In the case of paracrystalline rat red cells, which give spacings at 45 A and 58 A by x-ray diffraction, the specific heat is reduced to 85 per cent of that of the normal red cells, and the heat of compression is only about 75 per cent of that found for the normal red cell. In the case of the red cell sickled by a reduction of the O2 tension, the specific heat and the heat of compression are substantially the same as found for the normal red cell. This is an argument against sickling being the result of a crystallization process, and supports the observation that sickled cells do not give x-ray spacings. The result is compatible, on the other hand, with sickling being the result of the formation of an oriented and birefringent gel.  相似文献   

13.
Normal adult Sprague-Dawley rats were made anemic by repeated phlebotomy. Ion-exchange chromatography of anemic blood showed newborn like hemoglobin proportions, involving the same six hemoglobin components as is found when newborn and adult blood are compared. However, acetylsalicylic acid intake during anemia failed to demonstrate the changes in hemoglobin proportions, either totally or partially, depending upon the doses. Since acetylsalicylic acid inhibits prostaglandin synthesis, the data suggest that one or more prostaglandins may be involved in the process of reverse switching of hemoglobin in adult rat erythroid cells during erythropoietic stress.  相似文献   

14.
The hemoglobin minor/hemoglobin major ratio expressed in mouse erythroleukemia (MEL) cells grown in vitro varies according to the differentiation inducer utilized. For example, butyrate and hemin induce higher hemoglobin minor/hemoglobin major ratios than do dimethyl sulfoxide (DMSO) or hexamethylene bisacetamide (HMBA). Benzyl alcohol in non-toxic concentrations was found to markedly reduce the hemoglobin minor/hemoglobin major ratio and to moderately reduce the total hemoglobin induced by DMSO or HMBA in MEL cells, while only slightly decreasing the ratio induced by hemin or butyrate. The addition of dexamethasone (another and more potent inhibitor of the induction of hemoglobin synthesis than benzyl alcohol) to the media during HMBA induction of differentiation increased the hemoglobin minor/hemoglobin major ratio. This is similar to other "inhibitory" treatments (i.e., treatments that result in sub-optimal hemoglobin production) that have been previously reported. Therefore, although benzyl alcohol and dexamethasone both partly inhibit the induction of total hemoglobin production, they have opposite effects on the induced hemoglobin phenotype: benzyl alcohol decreases the hemoglobin minor/hemoglobin major ratio while dexamethasone increases it. The mechanism(s) of the alteration in the hemoglobin phenotype is unknown as is the mechanism of induction by any of the various inducing agents or of the inhibition of induction by any treatment. However, it appears that if the signal for the induction of hemoglobin minor is sufficiently potent (as it is during butyrate or hemin induction), it cannot be overcome by benzyl alcohol at a "non-toxic" concentration.  相似文献   

15.
1. A hemoglobin elution-staining procedure has been developed for distinguishing embryonic chick red blood cells from adult chicken red blood cells. 2. Adult hemoglobin is eluted from red blood cells with 1.9 M potassium phosphate buffer, pH 7.2; whereas, embryonic hemoglobin is retained within the cells and gives positive staining with erythrosin B. 3. The hemoglobin elution-staining pattern during development can be correlated with two embryonic hemoglobins as detected by polyacrylamide gel electrophoresis. 4. The series of red blood cells staining with erythrosin B correspond to the primary erythrocyte series suggesting that hemoglobin expression during development is correlated with different cell populations.  相似文献   

16.
A selective staining of hemoglobin in erythroid cell series was achieved by use of Sudan Black B (modified method of Sheehan and Storey) if optimal amount of hydrogen peroxide was added to the staining mixture. The effect of some inhibitory agents (KCN, wet heat, pH) on this staining as well as on the Lepehne's pseudoperoxidase reaction for hemoglobin was similar. Both reactions were more resistant to these factors than the peroxidase reactions and sudanophilia in granulocytes in which both could be blocked by the pretreatment with absolute methanol. Moreover the effect of some extraction procedures for lipids on both myeloperoxidase reactions and sudanophilia was investigated. The results support the view that the sudanophilia in granulocytes is due to their peroxidase activity and for the staining of hemoglobin by use of Sudan Black B with H2O2 its pseudoperoxidase activity is responsible. In addition the effect of the substitution of phenolphosphate by dihydroxybenzenes on granulocyte sudanophilia is reported.  相似文献   

17.
Two high precision techniques, titration microcalorimetry and thin-layer optical binding measurements, have made possible the evaluation of enthalpy changes for the overall oxygenation reactions for human hemoglobin (HbAo). Although the heat of adding three oxygen molecules could not be evaluated due to the indeterminate contribution of this species to the oxygen binding curve of the protein (Gill, S. J., Di Cera, E., Doyle, M. L., Bishop, G. A., and Robert, C. H. (1987) Biochemistry, 26, 3995-4002), the heats for binding two and four oxygen molecules were found to be simple multiples of the first binding heat. A direct consequence of equal stepwise heats is invariance of the shape of the binding curve with temperature, as pointed out by Wyman (Wyman, J. (1939) J. Biol. Chem. 127, 581-599). Titration microcalorimetry was also performed for the binding of carbon monoxide to hemoglobin. While the tight binding of CO precludes high-precision binding measurements, it does allow one to accurately determine the heat of ligation as a function of the CO bound. In these titrations a uniform heat of reaction is not observed, but the heat of binding increases markedly near the end point. This implies that the stepwise binding enthalpy for adding the third CO molecule is anomalously endothermic and for adding the fourth strongly exothermic. A similar phenomenon cannot be ruled out in the case of oxygen because of imprecision intrinsic in the analysis of the weaker ligand binding.  相似文献   

18.
1. The oxygen binding properties of the hemoglobin from the Lesser Rorqual, Balaenoptera acutorostrata, has been investigated with respect to the possible effects of organic phosphates on gas transport in arctic environments. 2. The intrinsic oxygen affinity of the hemoglobin is high and strongly modulated by the effects of organic phosphates. 3. In the absence of organic phosphates, the temperature sensitivity of oxygen binding expressed by the heat of oxygenation, delta H, is -16.2 kcal/mol when corrected for the heat of oxygen in solution. 4. In the presence of organic phosphates there is a marked decrease in the temperature sensitivity delta H approximately -5 kcal/mol). 5. This feature is of great importance for oxygen unloading in the flippers and the tail, where the temperature is lower than the trunk of the whale. 6. Furthermore the organic phosphates strongly increase the Bohr coefficient, delta log P50/delta pH, from less than -0.3 in stripped hemoglobin to about -1.5 when the hemoglobin is saturated with P6-inositol. 7. This feature may be of great physiological importance by reducing the CO2 tension and acidosis after a prolonged dive.  相似文献   

19.
A useful technique in studying the saturation of hemoglobin in erythrocytes or myoglobin in tissue is cryophotometry, in which tissue is frozen for later spectrophotometric analysis. A general question associated with this technique is whether the freezing process alters the chemical state. This paper presents a theoretical analysis of the simplest model relevant to that question. We study the effect of rapid cooling on a spatially homogeneous chemical reaction. The analysis shows that changes during freezing are negligible near the boundary to which the heat sink is applied, but can be significant deeper in the sample. The distance from the boundary at which the changes during freezing become appreciable can be expressed simply in terms of the chemical reaction rates and the thermal diffusivity of the tissue. Detailed results are given for the case of oxygen and myoglobin in skeletal muscle.  相似文献   

20.
Measurements of oxygen binding to bovine hemoglobin have been carried out over the temperature range 15-37 degrees C at pH 7.33. The standard enthalpy of oxygenation after correction for the heat of oxygen solution and of the Bohr protons is found to be -7.1 or -7.2 kcal/mol in the presence of 0.1 M chloride or bromide, respectively. This value is well below the -14.4 kcal/mol determined for human hemoglobin under identical experimental conditions. As reported by Fronticelli et al. (C. Fronticelli, E. Bucci and A. Razynska, J. Mol. Biol. 202 (1988) 343), the preferential binding of anions by bovine hemoglobin recognizes the various halides. Measurements at various temperatures reveal that this is true only above 25 degrees C. The halide recognition and the less exothermic enthalpy of oxygenation of bovine hemoglobin are probable due to oxygen-linked hydrophobic effects that are larger in bovine than in human hemoglobin.  相似文献   

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