[Effect of calcium hydroxide based desensitizing agent on cast crown retention].

A layer of desensitizing agent for prepared teeth was applied to tooth stump prior to the final cementation of cast crowns in order to alleviate postoperative sensibility. The aim of the study was to establish under in vitro conditions whether the application of the suspension of calcium hydroxide (Kaviner) on prepared teeth decreased the cast crown bonding strength. 40 intact human premolars were extracted for orthodontic reasons and prepared by high capacity machine using water supply cooling. Experimental crowns were made by a standard procedure using NiCrMo alloy and cemented by zinc phosphate or glass ionomer luting cements (20 in each group). Bond strength between prepared teeth and crowns was mechanically tested by using electronic dynamometer, at first without suspension of calcium hydroxide and then after its application. The results obtained pointed to decrease of bonding strength with both used luting cements: decrease of retention force with zinc phosphate cement was equal to 1.9% and with glass ionomer cement - 4.2%. However, the retention forces values obtained after the application of Kaviner for both dental cements observed were still clinically acceptable.

Inducible nitric oxide synthase in the myocard.

Recognition of significance of nitric oxide synthases (NOS) in cardiovascular regulations has led to intensive research and development of therapies focused on NOS as potential therapeutic targets. However, the NOS isoform profile of cardiac tissue and subcellular localization of NOS isoforms remain a matter of debate. The aim of this study was to investigate the localization of an inducible NOS isoform (NOS2) in cardiomyocytes. Employing a novel immunocytochemical technique of a catalyzed reporter deposition system with tyramide and electron microscopical immunocytochemistry complemented with Western blotting and RT-PCR, we detected NOS2 both in rat neonatal and adult cultured cardiomyocytes and in the normal myocard of adult rats as well as in the human myocard of patients with dilative cardiomyopathy. NOS2 was targeted predominantly to a particulate component of the cardiomyocyte--along contractile fibers, in the plasma membrane including T-tubules, as well as in the nuclear envelope, mitochondria and Golgi complex. Our results point to an involvement of NOS2 in maintaining cardiac homeostasis and contradict to the notion that NOS2 is expressed in cardiac tissue only in response to various physiological and pathogenic factors. NOS2 targeting to mitochondria and contractile fibers suggests a relationship of NO with contractile function and energy production in the cardiac muscle.

Effects of nitric oxide donors on energy metabolism of rat erythrocytes.

It is known that nitric oxide (NO) influences the activities of glycolytic enzymes, resulting in alteration of glycolysis rate. We investigated the effect of NO donors on the energy metabolism of rat erythrocytes. Rat erythrocyterich blood suspensions were aerobically incubated for 2 hours (1) as controls or (2) with different concentrations of NO donors: nitroglycerin (NTG), isosorbide dinitrate (ISDN), molsidomine (MO), and sodium nitroprusside (SNP). NTG, ISDN, MO, and SNP significantly (p < 0.05) increased glucose consumption and lactate accumulation in rat erythrocytes in a dose-dependent manner, indicating stimulation of glycolysis. The increased rate of glycolysis was accompanied by elevation of energy production (p < 0.05), but no changes in ATP levels were observed. The dose-dependent increase of glycolytic ATP production and the unaltered levels of ATP resulted in considerably shortened ATP-turnover time with the maximal experimental doses of NO donors used, indicating the stimulation of the ATP-consuming process in rat erythrocytes. The metabolic effects of NTG, ISDN, MO, and SNP were not mimicked by exogenous 8-Br-cGMP, NaNO2, or NaNO3, suggesting that the NO donor-induced stimulation of glycolysis and abbreviation of ATP-turnover time in rat erythrocytes was mediated by NO as an effector molecule, irrespective of the irreversible scavenger effect of hemoglobin. The implications of the NO effect on energy metabolism of erythrocytes is discussed.

Interaction between L-arginine: no system and cyclooxygenase metabolic products of arachidonic acid in coronary autoregulation.

Coronary autoregulation (CA) is the intrinsic ability of the heart to maintain its nutritive blood supply constant over a wide range of perfusion pressure. This phenomenon is regulated through several control mechanisms, while metabolic and myogenic control mechanism have dominant effects. In last few years, endothelial control mechanism, which is part of metabolic control, was intensive investigated. Dominant topic of endothelial-investigation was bioregulatory L-arginine: NO system, with his effective product--nitric oxide (NO). On the other hand, cyclooxygenase metabolic pathway products of arachidonic acid plays an important role in the control of vasomotor tone of coronary arteries. For this purpose, the aim of our study was to evaluate role of L-arginine: NO system, cyclooxygenase metabolites of arachidonic acid, as well as, their interactions in the control of CA of the isolated rat heart.. In our study rat hearts autoregulate CF between 50 and 90 cm H2O of CPP. Basal release (at 60 cm H2O) of NO (as nitrite), cAMP, cGMP and HX+X (i.e. adenosine) amounted to 2.85+/-0.25 nmol/min/g wt, 29.45+/-2.22 pmol/min/g wt, 0.43+/-0.08 pmol/min/g wt and 37.50+/-2.89 nmol/min/g wt respectively. Release of NO, cAMP and cGMP were strictly parallel with CPP-CF curve, while release of adenosine (i.e. HX + X) was an inverse function of perfusion pressure. Inhibition of NOS (L-NAME, 30 micromol/l) significantly widened autoregulatory range (40-100 cm H2O), with significant reduction in CF and NO- and cGMP release, while release of cAMP was completely reversed in the presence of L-NAME. However, inhibition of cyclooxygenase didn't influence autoregulatory range, with similar changes of NO- and cAMP-release and completely inversed values of released adenosine. When L-NAME an indomethacin (an nonspecific COX-inhibitor), 3 micromol/l where added together, they exhibit interactions between these two enzymatic systems. Namely, when L-NAME was added first, indomethacin didn't influence hemodynamic effects of NOS-inhibitor. On the other hand, when COX-inhibitor was added first, L-NAME widened autoregulatory range in small manner as after control autoregulatory experiments (40-90 cm H2O). All hemodynamic changes were followed with similar changes in NO-release, what suggest that exist interaction between L-arginine: NO system and COX-metabolites in the regulation of coronary autoregulation.

Effects of nitroglycerin on energy metabolism of rat reticulocytes.

Nitric oxide (NO) in many cells inactivates aconitase and mitochondrial respiratory chain, and influenced glyceraldehyde 3-phosphate dehydrogenase activity. The aim of this study was to evaluate role of nitroglycerin (NTG), a widely used NO donor, on energy metabolism of rat reticulocytes. Rat reticulocyte rich red blood cell suspensions containing 70-100% of reticulocytes, were aerobically incubated without (control) or in the presence of different concentrations of (a) NTG (0.1, 0.25, 0.5, 1.0, 1.5 mmol/l), (b) 8-Br-cGMP (0.1, 0.5, 1.0 mmol/l) and (c) NaNO2 and NaNO3 (1 mmol/l). NTG in dose- and time-dependent manner decreased total (p>0.05; EC50 = 0.78+/-0.05 mmol/l) and coupled (p<0.05; EC50 = 0.50+/-0.04 mmol/l) and increased uncoupled oxygen consumption (p<0.05: EC50 = 0.36+/-0.01 mmol/l). They were accompanied by stimulation of glycolysis, as measured by increased glucose consumption and lactate accumulation (p<0.001 EC50 = 0.53 and 0.53 mmol/l, respectively). Levels of all glycolytic intermediates in the presence of NTG indicate stimulation of HK-PFK, GA3PDH and PK activity. NTG significantly decreased ATP level, which accompanied by increased ADP and AMP levels. However, level of total adenine nucleotides (TAN) was significantly lower, which was consequence of increased catabolism of adenine nucleotides (increased hypoxanthine level; p<0.05). Stimulation of glycolysis accompanied with inhibition of the OxP, activation of HK-PFK, decrease of ATP and simultaneous rise of ADP and AMP levels, all together represent an example of Pasteur effect occurring in NTG-treated reticulocytes. In rat reticulocytes under steady state conditions 93% of overall energy was produced by OxP, but only 7% by glycolysis. Due to decrease of coupled oxygen consumption in the presence of NTG, ATP production via OxP was significantly diminished. Simultaneous increase of glycolytic ATP production is not enough to provide constant either ATP production or concentration. Calculated mean ATP-turnover time was prolonged even for 45% in the presence of 1.5 mmol/l NTG. Metabolic effects of NTG were not mimic by exogenous 8-Br-cGMP, NaNO2 or NaNO3, which indicate that NTG induced a) inhibition of coupled respiration and b) stimulation of glycolysis in rat reticulocytes are mediated by NO as an effector molecule.

Effects of nitric oxide donor, isosorbide dinitrate, on energy metabolism of rat reticulocytes.

Since nitric oxide (NO) in many cells is involved in energy metabolism, the aim of this study was to evaluate the role of isosorbide dinitrate (ISDN), a NO donor, in energy metabolism of rat reticulocytes, particularly due to their high content of hemoglobin--an effective scavenger of NO. Rat reticulocyte-rich red blood cell suspensions were aerobically incubated in the absence (control) or in the presence of different concentrations of ISDN. ISDN decreased total and coupled oxygen consumption (p<0.05) while increased uncoupled oxygen consumption (p<0.05) in a dose- and time-dependent manner. This was followed by enhancement of glycolysis, as measured by increased glucose consumption and lactate accumulation (p<0.05). Levels of all glycolytic intermediates in the presence of ISDN indicate only stimulation of pyruvate kinase activity. ISDN did not alter the concentration of ATP, while increased ADP and AMP levels (p>0.05). In rat reticulocytes under steady-state conditions, 95.4% of overall energy was produced by oxidative phosphorylation but only 4.6% by glycolysis. Due to a reduced coupled oxygen consumption in the presence of ISDN, ATP production via oxidative phosphorylation was significantly diminished. A simultaneous increase of glycolytic ATP production is not enough to ensure constant ATP production. The calculated mean ATP turnover time was prolonged by 199% in the presence of 1.5 mmol/l ISDN. In conclusion, ISDN a) inhibited total and coupled respiration but enhanced uncoupled respiration, b) stimulated glycolysis, c) decreased ATP production and d) prolonged ATP turnover time in rat reticulocytes. These effects were mediated by NO as the effector molecule.

The effect of cadmium and selenium on the antioxidant enzyme activities in rat heart.

Two month-old Wistar male albino rats were exposed during a 30-day period to a daily oral intake ad libitum of either 200 microg/mL Cd (as CdCl2), 0.1 microg/mL Se (as Na-selenite), or the same dosages of Cd + Se in drinking water. The daily intake from the water was calculated to be 15 mg Cd/kg and 7 microg Se/kg. Cadmium (Cd) accumulates in the heart (p < 0.005) and, in rats, decreases both body mass growth (p < 0.005) and heart mass (p < 0.02). Selenium (Se) significantly decreases the negative effect of Cd on body mass growth. In the hearts of Cd-treated rats, cadmium caused the decrease (p < 0.05) of selenium-dependent glutathione peroxidase (GSH-Px, EC 1.11.1.9) activity. At the same time, the activities of total superoxide dismutase (total SOD, EC 1.15.1.1), manganese-containing superoxide dismutase (Mn SOD), and copper-zinc-containing superoxide dismutase (CuZn SOD) were increased (p < 0.005). The activities of total SOD, CuZn SOD (p < 0.005), GSH-Px (p < 0.02), and glutathione-S-transferase (GST, p < 0.005) were increased in the hearts of Se-treated rats. However, by concomitant administration of Cd and Se, these changes were diminished (total SOD, GST) or were completely eliminated (Mn SOD, GSH-Px). These results indicate that Se only partly diminishes the effects of Cd cardiotoxicity.

Effect of cadmium and selenium on the antioxidant defense system in rat kidneys.

To examine effects of exogenous Cd on the kidney antioxidant defense system (AOS) and the possible protective role of Se against Cd toxicity, male Wistar albino rats (2 months old) were exposed during 30 days to oral intake of 200 ppm Cd (as CdCl2), 0.l ppm Se (as Na-selenite) or to the same doses of Cd / Se, simultaneously. Marked accumulation of Cd (23.44 +/- 0.69 micrograms/g w.m.) and marked alterations of AOS, resulting in kidney injury (renal pseudohypertrophy), were found in Cd-treated rats. Activities of total superoxide dismutase (SOC, EC 1.15.1.1), manganese-containing superoxide dismutase (MnSOD) and selenium-dependent glutathione peroxidase (Se GSH-Px, EG 1.11.1.9) were significantly reduced, whereas that of glutathione-S-transferase (CST, EC 2.5.1.18) and vitamin E (vit E) concentration were significantly increased in the kidneys of Cd-treated rats. Kidney catalase (CAT, EC 1.11.1.6) activity, ascorbic acid (AsA) and red blood cell glutathione (GSH, GSSG) levels were not markedly influenced by CD uptake. In kidneys of Se treated rats, the activities of total SOD, copper-zinc-containing superoxide dismutase (CuZnSOD) and GST were significantly increased Activities of kidney CAT and Se GSH-Px were largely unchanged, whereas significant increases of the kidney AsA and vit E concentrations occurred. In Cd + Se-cotreated rats, the kidney activities of MnSOD, CAT and Se GSH-Px, as well as vit E concentration, were the same as in controls, whereas CuZnSOD and GST activities and concentration of AsA exceeded normal values. These data indicate that Se only partially improves the AOS that is insufficient to prevent Cd-induced nephrotoxicity.

Intracellular localization of inducible nitric oxide synthase in neonatal rat cardiomyocytes in culture.

Recognition of the role of nitric oxide (NO) in cardiovascular regulations raised an acute interest in NO-generating enzymes-nitric oxide synthases (NOS). Nevertheless, the subcellular localization of inducible isoform of NOS (NOS II) and regulation of its expression in the cardiomyocyte still remains to be elucidated. Therefore, we focused this study on the subcellular localization of NOS II in cultured neonatal rat cardiomyocytes using immunocytochemical techniques at the light and electron microscopic level as well as the demonstration of NADPH-diaphorase activity and the Griess assay for NO measurement. Cultivation of neonatal cardiomyocytes during 2 and more days induced a moderate increase in the NOS II immunolabeling in defined cytoplasmic structures and a nuclear NOS II staining in some cells. Exposure of the cell cultures to exogenous cAMP markedly stimulated NO production with a concomitant enhancement of NOS II immunolabeling of cardiomyocytes. cAMP-induced changes were significantly attenuated by dexamethasone. This report provides evidence for the localization of NOS II in the perinuclear space, Golgi complex, mitochondria, plasma membrane and along contractile fibers of cardiomyocytes, as well as for the appearance of NOS II staining of the cell nuclei in the course of cultivation. In non-cardiomyocytes contaminating the cell culture, positive immunoreaction was detected in the Golgi complex and endoplasmic reticulum. Our data point to a notable constitutive expression of NOS II in rat cardiomyocytes apparently dependent on the developmental stage.

Altered expression of PDE1 and PDE4 cyclic nucleotide phosphodiesterase isoforms in 7-oxo-prostacyclin-preconditioned rat heart.

The stable prostacyclin derivative, 7-oxo-prostacyclin, exhibits a delayed, long-lasting cardioprotective effect, which is accompanied by an increase in cyclic nucleotide phosphodiesterase (PDE) activities restricted to the Ca2+-calmodulin-dependent (PDE1) and cyclic AMP-specific phosphodiesterase (PDE4) activities. Mammalian PDEs form a large multi-gene family with differential expression occurring in a cell- and tissue-specific manner. The aim of this study was to identify which isoforms of PDE1 and PDE4 are present in the hearts of control and 7-oxo-prostacyclin treated rats. Using RT-PCR analysis, we were able to identify in control rat hearts transcripts for PDE1C, but not for either PDE1A or PDE1B within the three-gene PDE1 family. Within the four-gene PDE4 family we detected, by generic RT-PCR analysis, transcripts for PDE4A, PDE4B and PDE4D, but not PDE4C. Using RT-PCR primers for specific splice variants, we identified transcripts for PDE4B1, PDE4B2, PDE4B3, PDE4D1, PDE4D2 and PDE4D3 in hearts from the control animals. Immunoblotting of hearts from the control animals for PDE4 forms allowed us to identify a 98-kDa PDE4A species, 68-kDa band representing PDE4D1/2 and a 95-kDa species indicative of PDE4D3. In the hearts of treated animals, 48 h after a single 50 microgram/kg dose of 7-oxo-prostacyclin, a profound increase in transcript levels was seen for both PDE1C and PDE4B3 together with a slight elevation for PDE4B1. No change in PDE4A transcripts occurred, which was consistent with a lack of change indicated in immunoblotting analyses. In contrast, 7-oxo-prostacyclin treatment caused decrease in transcript levels for PDE4D, which was confirmed by immunoblotting and shown to be due to a reduction in the levels of PDE4D3 and also in PDE4D1/D2. Thus, treatment of animals with 7-oxo-prostacyclin initiated profound isoform-specific changes in PDE expression in the myocardium which, presumably, underpin the increased PDE activity.

Alteration of erythrocyte membrane Na, K-ATPase in children with borderline or essential hypertension.

The aim of this study was to evaluate the substrate (ATP) kinetics of erythrocyte membrane Na, K-ATPase in children with borderline or essential hypertension. Although the activity of Na, K-ATPase in the presence of in vivo concentrations of ATP was not significantly altered, kinetic studies showed an obvious inhibition of enzyme activity in the erythrocyte membrane of children with borderline or essential hypertension. Hanes plot analysis revealed a decrease of V(max) from 7.19 in erythrocytes from control subjects to 4.93 and 3.33 in those from children with borderline or essential hypertension, respectively. A mean value of the K(m) decreased from 0.10 in the control to 0.08 and 0.02 in children with borderline or essential hypertension, respectively. The energy status of erythrocytes, estimated by ATP, ADP and AMP levels, ATP/ADP ratio, and adenylate energy charge (AEC) was not significantly changed in the cells from hypertensive children. The use of a free radical-generating system (FeSO4/ascorbate) in vitro significantly reduced enzyme activity in the control erythrocytes while in those from hypertensive children it was abolished completely. The level of lipid peroxides was considerably higher (+ 37 per cent) in the plasma, while that of reduced glutathione was significantly lower both in the erythrocytes and the plasma of children with essential hypertension than in healthy children. These results indicate significant alterations of the antioxidant status which could be the cause of the inhibited Na, K-ATPase activity in erythrocyte membranes from hypertensive children.

Role of nitric oxide (NO) in the regulation of coronary circulation.

Nitric oxide plays an important role in the control of basal coronary tone and mediation of reactive hyperaemic flow response following short-term coronary occlusion. The results presented in this report indicate that NO is involved in the modulation of coronary autoregulation in isolated rat hearts. Isolated rat heart exhibit autoregulation of coronary flow (CF) between 50 and 80 cm H2O of coronary perfusion pressure (CPP). Within this autoregulatory range NO release (measured as nitrite) varies from 1.7 +/- 0.3 to 2.2 +/- 0.7 nmol/min/g wt. Below the autoregulatory range it decreases slightly, while above this there is more than a twofold increase. Changes of NO release are accompanied by directly proportional changes of cGMP release. The release of hypoxanthine + xanthine shows a reciprocal relationship to CF values. The inhibition of NO synthesis showed a reciprocal relationship with CF values. Inhibition of NO synthesis by L-NAME (30 mumol/l) significantly reduces CF over the entire range of CPP changes (20-120 cm H2O), but much less at lower than at higher pressure values. Therefore, the autoregulatory range is significantly widened to CPP of 40-100 cm H2O. Theophylline (30 mumol/l) reduces CF by 15-25% throughout the entire range of CPP changes. Hence, the CPP-CF curve is shifted downwards without significant changes of the autoregulatory range. Theophylline-induced reduction of NO release is CPP-dependent: as greater as CPP lower. When L-NAME is coadministered with theophylline, CF is additionally reduced while widened autoregulatory range is shifted to the right.

Role of histamine in the regulation of coronary circulation.

The aim of this study was to evaluate the role of endogenous histamine in the regulation of reactive hyperaemia (RH) and coronary autoregulation in isolated rat hearts. The basal release of cardiac histamine (perfusion pressure 60 cm H2O) amounted to 100-200 pmol/min/g wt. During the first 15 s following 30 s of coronary occlusion, the release of histamine increased about three times and returned to basal levels after approximately 90 s, paralleling the changes of coronary flow (CF). Blockade of H1-receptors increased basal CF by 23 +/- 2%, significantly reduced blood flow debt and prolonged the duration of RH. Blockade of H2- and H3-receptors produced a significant decline of CF, decreased RH flow and diminished RH by 40 +/- 3%. Blockade of all three classes of histamine receptors indicated that endogenous histamine exerts predominantly vasodilatory effects (mediated by H2- and H3-receptors) on coronary circulation. Histamine-induced vasodilation appears to be NO-dependent. Changes of coronary perfusion pressure from 20 to 120 cm H2O were accompanied by an almost linear decrease of histamine release from about 200 to 45-50 pmol/min/g wt. Blockade of histamine receptors decreased, while L-NAME significantly widened the autoregulatory range of the isolated rat heart, reduced CF and release of NO, but reversed the pattern of histamine release leaving the autoregulatory range unaltered, which indicate that endogenous histamine does not play a role in the regulation of coronary autoregulation.

Biphasic L-arginine uptake by the isolated guinea-pig heart.

L-Arginine is the physiological substrate for the formation of nitric oxide (NO) and accounts for the biological activity of endothelium-derived relaxing factor. We have studied L-arginine transport in the heart using a rapid dual-isotope dilution technique. The time course of L-[3H]arginine uptake (extraction) by the isolated perfused guinea-pig heart was found to occur in two phases. The first phase reached a plateau in 6.6 +/- 0.6 s and lasted 8.8 +/- 0.7 s, whereas the second phase developed a plateau after 16.3 +/- 0.8 s. The first phase of maximal uptake (Umax,1) accounted for 13.4 +/- 1.4% of the total uptake and the second (Umax,2) for 32.3 +/- 1.8%. The two phases of uptake were inhibited by unlabelled L-arginine in a dose-dependent manner, which suggests that both phases are carrier mediated. The degree of inhibition of Umax,1 and Umax,2 by unlabelled L-arginine was not significantly different. Studies of the kinetics of uptake of these processes revealed an apparent Km,1 of 183 +/- 10 microM with a Vmax,1 of 50 +/- 10 nmol min-1 g-1 for the first phase and Km,2 of 167 +/- 14 microM with a Vmax,2 of 93 +/- 13 nmol min-1 g-1 for the second phase of uptake. These results suggest a similar affinity for the receptors of both transport systems, but with different values for Vmax (P < 0.05). In contrast, 1 mM unlabelled D-arginine had no effect on either the first or second phase of uptake of L-[3H]arginine by the heart, which suggests that these processes are stereospecific. In the presence of the L-stereoisomer of nitro-arginine-mono-methyl ester (L-NAME), a potent inhibitor of NO synthesis, the Umax,1 was inhibited by about 60% while Umax,2 was inhibited by only 20%, which suggests that there is a difference in the effect of L-NAME on the two phases of L-arginine uptake. The first phase most probably represents uptake into the capillary wall, i.e. endothelium and smooth muscle, while the second phase represents entry into the extra-endothelial compartment, i.e. the cardiac myocytes and fibroblasts.

Interplay of nitric oxide and histamine in the regulation of coronary reactive hyperemia and coronary autoregulation.

Reactive hyperemia following 30 s of coronary occlusion in the isolated guinea pig heart is accompanied by a two-fold increase of nitric oxide (NO) and histamine release, which are significantly reduced in the presence of L-NAME, cimetidine and thioperamide, respectively. Great changes of histamine release occur during autoregulation. However, histamine seems much more important for metabolic dilation below the autoregulatory range. Inhibition of NO synthesis, but not blockade of histamine receptors, widens the autoregulatory range. Changes of the released NO and histamine under conditions employed in this study suggest a positive feed-back relationship between NO and histamine in the regulation of coronary circulation.

The effects of selenium on the antioxidant defense system in the liver of rats exposed to cadmium.

Total superoxide dismutase (total SOD), copper zinc containing superoxide dismutase (CuZn SOD), and manganese superoxide dismutase (Mn SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione-S-transferase (GST) activities as well as ascorbic acid (AsA), and vitamin E (vit E) concentrations were analysed in the liver of rats exposed to cadmium (15 mg Cd/day/kg), selenium (7 micrograms Se/day/kg), and to cadmium+selenium (15 mg Cd + 7 micrograms Se/day/kg), and in control animals. Cadmium caused a decrease of total SOD, Mn SOD, CAT and GSH-Px but an increase of GST activity in the liver of rats. Contrary to cadmium, selenium caused a significant increase of the activity of these enzymes except for GSH-Px. By concomitant exposure to both cadmium and selenium, the toxic effects of cadmium on the activity of mentioned enzymes we abolished. In all exposed groups, the activity of enzyme glutathione-S-transferase was enhanced, indicating its increased role in prevention of lipid peroxidation. Cadmium decreased the concentration of AsA and increased the concentration of vitamin E in the liver, while selenium increased the concentration of both vitamins. However, by concomitant administration of cadmium and selenium, these changes were diminished and tended to reach control values.

Energy metabolism of reticulocytes: two different sources of energy for Na+K(+)-ATPase activity.

Total energy production in rabbit reticulocytes amounted to 136.52 +/- 6.50 mumol ATP h-1 ml-1 of reticulocytes: 88.3 per cent was provided by oxidative phosphorylation, whereas only 11.7 per cent by aerobic glycolysis. Na+K(+)-ATPase accounted for 23 per cent, i.e. 27.65 +/- 2.55 mumol ATP h-1ml-1 of reticulocytes, in the overall energy consumption in reticulocytes of rabbits. Under basal conditions ATP for Na+K(+)-ATPase activity was derived exclusively from oxidative phosphorylation. However, when the activity of Na+K(+)-ATPase was increased due to the stimulation of adenylate cyclase by (-)-isoprenaline, the additional energy required was provided by aerobic glycolysis. These results indicate that two different compartments, one cytosolic and the other mitochondrial, provide energy for Na+K(+)-ATPase activity in reticulocytes.

Cadmium-induced changes of antioxidant and metabolic status in red blood cells of rats: in vivo effects.

Chronic exposure of adult rats to dietary intake of cadmium (15 mg CdCl2/day/kg for 30 days) leads to development of anemia and thrombocytosis. Anemia is characterized by significant reticulocytosis (13.1 +/- 1.0%), anysocytosis, poikilocytosis, iron deficiency and marked alterations of antioxidant and metabolic status of red blood cells. Activities of SOD, catalase, GPx and GR were significantly increased in red blood cells of cadmium-treated rats. In treated animals cadmium induced an increase of red cell reduced and oxidized glutathione with no changes of GSSG/GSH ratio. However, significant reduction of lipid peroxidation was found. Plasma levels of tocopherol and ascorbate, as well as activity of glutathione-S-transferase, were all significantly increased in cadmium-treated rats. The energy metabolism of red blood cells was deeply altered in cadmium-treated rats. The levels of ATP, ADP, AMP and TAN were significantly increased while ATP/ADP ratio and adenylate energy charge (AEC) were significantly reduced. The level of 2,3-BPG was somewhat lower, but 2,3-BPG/Hb ratio was considerably higher, in red blood cells of cadmium-treated rats.

Role of nitric oxide in reactive hyperemia of the guinea pig heart.

To evaluate the role of nitric oxide (NO) in the flow response after brief coronary arterial occlusion, NO formation by the isolated guinea pig heart was assessed by a specific difference spectrophotometric assay. Release of NO under basal conditions was 121.8 +/- 10.5 pmol x min-1 and increased to 211.1 +/- 16.8 pmol x min-1 after 60 seconds of coronary occlusion. Simultaneously, release of cGMP and adenosine increased by 87% and 652%, respectively. The kinetics of NO release paralleled the reactive hyperemic flow response. Inhibition of NO synthesis with nitro-L-arginine methyl ester (L-NAME, 30 microM) significantly reduced basal flow and attenuated reactive hyperemia, flow repayment, and repayment ratio. L-NAME decreased release of cGMP but significantly increased adenosine release under basal conditions and during reactive hyperemia. Oxyhemoglobin (5 microM) potentiated the effects of L-NAME. The stereoisomer nitro-D-arginine methyl ester was ineffective. Our results suggest 1) NO is an important regulator of coronary flow during reactive hyperemia as well as under basal flow conditions and 2) the significance of the increased adenosine release when NO synthesis is inhibited remains to be determined.