Characteristics of cardiac rhythm and the peroxidase activity of blood in patients with ischemic heart disease treated by hyperbaric oxygenation]

The myeloperoxidase of peripheral blood neutrophils revealed by the cytochemical method was investigated in a group of patients with ischemic heart disease (24 men, mean age--51) treated with HBO (10 sessions, under 1.5-1.7 ata, for 60 min) prior to and after each HBO session. The ECG monitoring was performed during these sessions as well. A correlation analysis of the peroxidase index (mean enzyme content in 100 cells), total peroxidase activity and tension index (obtained by computer analysis of cardiac rhythm changes) was carried out in that group of patients. A close correlation was found between the total peroxidase activity and tension index.     

Hyperbaric oxygen preconditioning alleviates myocardial ischemic injury in rats

It has been shown that after ischemia-reperfusion, application of hyperbaric oxygen (HBO) reduces cardiac injury. In this study we tested the hypothesis that HBO preconditioning reduces injury to the ischemic myocardium. One hundred and eight adult male Sprague-Dawley rats (250-280 g) were randomly divided into four groups: normoxia + sham surgery (CS), normoxia + permanent occlusion of the left anterior descending (LAD) coronary artery (CMI), HBO preconditioning + sham surgery (HS), and HBO preconditioning + permanent LAD occlusion (HMI). Rats receiving HBO preconditioning were intermittently exposed to 100% O(2) at 2.5 atmosphere absolute (ATA) for 60 min, twice daily for 2 days followed by 12 hrs of recovery in room air prior to the myocardial ischemic insult induced by LAD ligation. Rats in the normoxia group were time-matched with the HBO group and maintained under normoxic conditions prior to LAD occlusion. At 3 and 7 days after LAD occlusion, heart function parameters were measured by inserting a catheter into the left ventricle, infarct size was calculated using the method of TTC staining, myocardial capillary density was determined by immunohistochemical staining with a monoclonal anti-CD(31)/PECAM-1 antibody, and VEGF protein level was determined by Western blot analysis. At 3 days after LAD ligation, the infarct size of the HMI group was significantly smaller than that of the CMI group (26 +/- 2.5% vs. 38 +/- 3%, P < 0.05). The heart function parameters including left ventricular systolic pressure (LVSP), +dP/dt(max) and -dP/dt(max) were significantly improved in the HMI group compared to the CMI group at 3 and 7 days after LAD occlusion. Capillary density and VEGF protein levels were significantly increased in the ischemic myocardium pre-exposed to HBO. We conclude that HBO preconditioning alleviates myocardial ischemia in rat model.     

Real-time monitoring of nitric oxide in ischemic myocardium using an NO-selective electrode calibrated by electron spin resonance

Using a Langendorff-perfused rat heart preparation and selective electrodes, we determined nitric oxide (NO) and oxygen levels in cardiac tissue. An NO-selective electrode that was calibrated by electron spin resonance (ESR) spectroscopy was inserted into the middle of the myocardium in the left ventricle. Simultaneously, we used an O2-selective electrode to measure the partial pressure of oxygen (pO2) in the perfusate, Krebs-Henseleit (K-H) solution, that was ejected from the heart. After 30 min of aerobic control perfusion, hearts were subjected to 30 min of global ischemia followed by 30 min of reperfusion. Under ischemic conditions, with a gradually decreasing pO2, NO detected by an NO-sensitive electrode within the myocardium was gradually increased. The maximum concentration increases in NO and decreases in pO2 during global ischemia were +10.200 +/- 1.223 microM and -58.608 +/- 4.123 mmHg, respectively. NO and pO2 levels both recovered to pre-ischemia baseline values when perfusion was restarted after global ischemia (reperfusion). The presence of Nomega-nitro-L-arginine methyl ester (L-NAME, 10 mM), a NOS inhibitor, prevented ischemia/reperfusion-induced changes in NO. This study shows that an NO-selective electrode that is calibrated by ESR can provide accurate, real-time monitoring of cardiac NO in normal and ischemic myocardium.     

HBO suppresses Nogo-A,Ng-R,or RhoA expression in the cerebral cortex after global ischemia

Nogo-A, a myelin-associated neurite outgrowth inhibitory protein, binds with the Ng-R receptor to activate RhoA intracellular signals and inhibit the plasticity after CNS injury. We evaluated the effect of hyperbaric oxygen (HBO) on the expression of Nogo-A, Ng-R, and RhoA after transient global ischemia in a rat 2 vessel occlusion global ischemic model. Male SD rats (n=78) were randomly divided into 13 groups: 1 sham group, 6 groups of global ischemia, and 6 groups of HBO treatment after global ischemia. HBO (3ATA) was applied for 2 hr at 1 hr after global ischemia. Rats were sacrificed at 6, 12, 24, 48, and 96 hr and 7 days. Global ischemia (10 min) produced a marked increase of Nogo-A/B, Nogo-A, Ng-R, and RhoA expression. Immunohistochemistry showed increased Nogo-A/B and Nogo-A located in the myelin sheath of ischemic brain cortex. Ng-R expressed on the surface of neurons and their processes, and RhoA expressed inside the cytoplasm of neurons in ischemic brain. HBO significantly reduced neurological injury, decreased the levels of Nogo-A, Ng-R, and RhoA in ischemic injured cortex (p<0.05).     

Genetic effects of hyperbaric oxygen therapy

Patients with several diseases have been examined for detection of chromosome aberrations in peripheral blood cells after 10 sessions of hyperbaric oxygen (HBO) at 0.15-0.20 MPa for 40 min. The present study reveals that HBO increases the level of chromosome aberrations, and that individual reactions to HBO differ. Pure erythrocytes treated with high-pressure oxygen (HBO) at 0.7 MPa for 1 h are clastogenic for intact syngeneic lymphocytes. The effect of HBO (0.3 MPa, 5 sessions of 1 h daily) on induction of chromosome aberrations in somatic cells and germinal tissues of rat males has been studied. Induction of aberrations in bone marrow cells after HBO was seen for 3 months. In lymphocytes of patients, it was seen for 9 months. Chromosome rearrangements at the first meiotic division were detected only 90 days after exposure. HBO affects neither the functional nor the morphological condition of gonads and does not induce dominant lethals. It is proposed that a high quantity of chromosome breaks in cells of somatic tissues is an adaptive reaction of organisms to HBO.     

Effect of ceruloplasmin on the oxygen tension in muscular tissue of experimental animals]

The polarographic method using platinum electrode has been applied to study the effect of ceruloplasmin (CP) on the oxygen tension (pO2), oxygen saturation rate and rate of oxygen utilization in the muscular tissue of high-leukemic AKR mice, C57BL/6 mice with transplanted Lewis lung carcinoma (3LL) and rats after gamma-irradiation in a dose of 7 Gr. It has been shown that CP in AKR mice improves oxygen saturation of the muscular tissue. This effect is also evident in the case of the marked pO2 decrease in the muscle and its oxygen saturation rate (animals with Lewis lung carcinoma and after gamma-irradiation).     

Endotoxin-induced liver hypoxia: defective oxygen delivery versus oxygen consumption.

In vivo EPR was used to investigate liver oxygenation in a hemodynamic model of septic shock in mice. Oxygen-sensitive material was introduced either (i) as a slurry of fine particles which localized at the liver sinusoids (pO2 = 44.39 +/- 5.13 mmHg) or (ii) as larger particles implanted directly into liver tissue to measure average pO2 across the lobule (pO2 = 4.56 +/- 1.28 mmHg). Endotoxin caused decreases in pO2 at both sites early (5-15 min) and at late time points (6 h after endotoxin; sinusoid = 11.22 +/- 2.48 mmHg; lobule = 1.16 +/- 0.42 mmHg). The overall pO2 changes observed were similar (74.56% versus 74.72%, respectively). Blood pressures decreased transiently between 5 and 15 min (12.88 +/- 8% decrease) and severely at 6 h (59 +/- 9% decrease) following endotoxin, despite volume replacement with saline. Liver and circulatory nitric oxide was elevated at these times. Liver oxygen extraction decreased from 44% in controls to only 15% following endotoxin, despite severe liver hypoxia. Arterial oxygen saturation, blood flow (hepatic artery), and cardiac output were unaffected. Pretreatment with l-NMMA failed to improve endotoxin-induced oxygen defects at either site, whereas interleukin-13 preserved oxygenation. These site-specific measurements of pO2 provide in vivo evidence that the principal cause of liver hypoxia during hypodynamic sepsis is reduced oxygen supply to the sinusoid and can be alleviated by maintaining sinusoidal perfusion.     

Cerebral blood flow and tissue pO2 changes following local met-enkephalin administration in awake, freely moving cats

The effect of locally administered methionine-enkephalin on cerebrocortical blood flow and tissue pO2 was tested in chronic, awake, freely moving cats. Local cortical blood flow was measured with the H2-gas clearance method, and cortical pO2 in the same area was monitored with a polarographic technique using double Pt electrodes. Met-enkephalin was administered with a micropipette technique in 0.1, 0.5, 1.0 and 5.0 mumol/l concentrations in 20 microliters volume. Met-enkephalin caused a marked, dose dependent decrease of the cortical flow as well as of the cortical pO2. Following 5.0 mumol/l met-enkephalin the flow decreased more (x = 30.6%) than did the pO2 (x = 18%). The maximum effect could be observed 2-7 min following administration of the peptide, the duration of the effect was about 20 min. The threshold dose was found around 5-7 X 10(-8) M/l. Met-enkephalin in the above mentioned doses caused no vegetative, motor or behavioural changes in these experiments.     

Hyperbaric oxygenation and drug therapy in treatment of nonspecific ulcerative colitis and Crohn's disease]

Patients with nonspecific ulcerative colitis and Crohn's disease were treated with drug therapy (prednisolone, sulphasalazine, metronidazole per os and hydrocortisone per rectum) and subjected to 12 sessions of HBO. Every 10-14 months the HBO course was repeated. After 7 years of such a treatment a gradual partial recovery of large intestine mucosa and in some cases even its absolute recovery were observed. It was most difficult to get good results when treating lesion of distal colon segments. HBO produced a good effect when the disease was diagnosed at the early stage, when the intestine injury was accompanied by hepatobiliary system disease and in teenagers.     

Preconditioning with hyperbaric oxygen induces tolerance against oxidative injury via increased expression of heme oxygenase-1 in primary cultured spinal cord neurons

Hyperbaric oxygen (HBO) preconditioning can induce ischemic tolerance in the spinal cord. The effect can be attenuated by the administration of an oxygen free radical scavenger or by inhibition of antioxidant enzymes. However, the mechanism underlying HBO preconditioning of neurons against ischemic injury remains enigmatic. Therefore, in the present study primary cultured spinal cord neurons were treated with HBO and then subjected to a hydrogen peroxide (H(2)O(2)) insult. The results show that H(2)O(2) stimulation of the cultured spinal neurons caused severe DNA damage and decreased cell viability, and that these neurons were well protected against damage after a single exposure to HBO preconditioning (0.35 MPa, 98% O(2), 37 degrees C, 2 h). The protective effect started 4 h after pretreatment and lasted for at least 24 h. The cultured neurons after HBO treatment also exhibited increased heme oxygenase-1 (HO-1) expression at both the protein and mRNA levels, which paralleled the protective effect of HBO. Treatment with tin-mesoporphyrin IX (SnMP), a specific HO-1 inhibitor, before HBO pretreatment abolished the HBO-induced adaptive protection noted in the cultured spinal neurons. In conclusion, HBO preconditioning can protect primary cultured spinal cord neurons against oxidative stress, and the upregulation of HO-1 expression plays an essential role in HBO induced preconditioning effect.     

Regional myocardial ischemia-induced activation of MAPKs is associated with subcellular redistribution of caveolin and cholesterol

Ischemia-reperfusion activates ERK and p38 MAPK in cardiac membranes, but the role of caveolae in MAPK signaling during this stress has not been studied. The purpose of this study was to determine the effect of in vivo myocardial ischemia-reperfusion on the level and distribution of caveolin-1 and -3 and cholesterol as well as MAPK activation in caveolin-enriched fractions. Adult male rats were subjected to in vivo regional myocardial ischemia induced by 25 min of coronary artery occlusion and 10 min (n = 5) or 2 h (n = 4) of reperfusion. Another group of rats served as appropriate nonischemic time controls (n = 4). A discontinuous sucrose density gradient was used to isolate caveolae/lipid rafts from ischemic and nonischemic heart tissue. Caveolin-1 and -3, as well as cholesterol, were enriched in the light fractions. A redistribution of caveolin-3 and a reduction in caveolin-1 and cholesterol levels in the light fractions occurred after 10 min of reperfusion. The ERKs were activated in ischemic zone light and heavy fractions by 10 min of reperfusion. p44 ERK was activated after 2 h of reperfusion only in the light fractions, whereas p42 ERK phosphorylation was increased in the light and heavy fractions. Although no p38 MAPK activation occurred after 10 min of reperfusion, 2 h of reperfusion caused significant activation of p38 MAPK in nonischemic zone light and heavy fractions. These results show the importance of caveolar membrane/lipid rafts in MAPK signaling and suggest that subcellular compartmentation of p44/p42 ERKs and p38 MAPK may play distinct roles in the response to myocardial ischemia-reperfusion.     

The effect of hyperbaric oxygenation on rat somatic and generative cells

The effect of higher oxygen pressures (0.3 MPa, 5 sessions 1 h each daily) on induction of chromosomal aberrations in somatic cells and germinal tissues of rat males has been studied. Appearance of aberrations in bone marrow cells after HBO was recorded for 3 months, in cells of eye cornea epithelium--during 3 weeks after the end of sessions. Chromosome rearrangements in the first meiotic division were detected only 90 days after sessions. HBO affects neither functional nor morphological conditions of gonads and induces no dominant lethals. It is suggested that mutagenesis in bone marrow cells is an adaptive process, preventing mutagenic consequences of HBO in the cells of germinal tissue.     

Method of isolation of intact enterocytes from rat duodenal mucosa

A procedure is suggested for obtaining intact enterocytes from rat duodenum mucosa by the tissue trypsinization in the isotonic Krebs solution (pH 7.9). The maximum yield of certain enterocytes is reached at the enzyme concentration of 0.03%; the incubation time--15 min; 200-250 mg of mucosa produces 3,7 X 10(6)--4,7 X 10(6) cells.     

Diurnal rhythm of acid-base equilibrium and blood gas composition

Diurnal fluctuations of acid-alkaline balance and blood gas composition were studied in March on 20 healthy males aged 20-26. Using cosinor analysis, mesors, amplitudes and acrophases were revealed. Calculated acrophases (in h and min) are as follows: pH--16 h 05 min, standard hydrogen carbonate--18 h 45 min, actual hydrogen carbonate--22 h 55 min, buffer bases--19 h 03 min, pCO2--2 h 47 min, pO2--4 h 39 min, HbO2--8 h 07 min, hemoglobin--14 h 16 min. All the values studied during diurnal cycle correlated, forming certain circadian rhythms.     

Hyperbaric oxygenation in the comprehensive therapy of patients with rheumatoid arthritis (clinico-immunologic study)

For 35 of 50 patients with rheumatoid arthritis traditional drug therapy was a minor success for a long time. Without any modifications of the drug therapy every patient went through a course of hyperbaric oxygenation (HBO): 21 sessions under 1.7 ata for 40 min. Good clinical results both immediate and remote have been obtained. The effect of HBO on the immune system of the patients has intensified the suppressive function of T-lymphocytes (especially with systemic symptoms of the disease), normalized cell-bound immunity and decreased the serum concentration in immune complexes.     

Protection of calcitonin gene-related peptide-mediated preconditioning against coronary endothelial dysfunction induced by reperfusion in the isolated rat heart

This study was designed to explore the protective effect of ischemic preconditioning on reperfusion-induced coronary endothelial dysfunction, with a focus on the role of calcitonin gene-related peptide (CGRP) in this effect, in the isolated perfused rat heart. Thirty minutes of global ischemia and 30 min of reperfusion significantly decreased heart rate, left ventricular pressure, and its first derivative and impaired vasodilator responses to acetylcholine. Ischemia-reperfusion did not affect vasodilator responses to sodium nitroprusside. Preconditioning induced by three cycles of 5 min of ischemia and 5 min of reperfusion produced a significant improvement in cardiac function concomitantly with an amelioration of vasodilator responses to acetylcholine. The protective effects of ischemic preconditioning were abolished by CGRP(8-37) (10(-7) M) , the selective CGRP receptor antagonist. After pretreatment with capsaicin (50 mg x kg(-1), s.c.) to deplete endogenous CGRP, the preconditioning effect was absent. Pretreatment with exogenous CGRP (5 x 10(-9) M) for 5 min induced a preconditioning-like protection. The present study suggests that the cardioprotection of ischemic preconditioning is related to the preservation of the coronary endothelial cell, and that the protective effect of preconditioning is mediated by endogenous CGRP in the isolated perfused rat heart.     

Reference cardiopulmonary values in normal dogs

The purpose of this project was to collate canine cardiopulmonary measurements from published and unpublished studies in our laboratory in 97 instrumented, unsedated, normovolemic dogs. Body weight; arterial and mixed-venous pH and blood gases; mean arterial, pulmonary arterial, pulmonary artery occlusion, and central venous blood pressures; cardiac output; heart rate; hemoglobin; and core temperature were measured. Body surface area; bicarbonate concentration; base deficit; cardiac index; stroke volume index, systemic and pulmonary vascular resistance indices; left and right cardiac work indices; alveolar partial pressure of oxygen (pO2) ; alveolar-arterial pO2 gradient (A-apO2); arterial, mixed-venous, and pulmonary capillary oxygen content; oxygen delivery; oxygen consumption; oxygen extraction; venous admixture; arterial and mixed-venous blood CO2 contents; and CO2 production were calculated. In the 97 normal, resting dogs, mean arterial and mixed-venous pH were 7.38 and 7.36, respectively; partial pressure of carbon dioxide (pCO2), 40.2 and 44.1 mm Hg, respectively; base-deficit, -2.1 and -1.9 mEq/liter, respectively; pO2, 99.5 and 49.3 mm Hg, respectively; oxygen content, 17.8 and 14.2 ml/dl, respectively; A-a pO2 was 6.3 mm Hg; and venous admixture was 3.6%. The mean arterial blood pressure (ABPm), mean pulmonary arterial blood pressure (PAPm), pulmonary artery occlusion pressure (PAOP) were 103, 14, and 5.5 mm Hg, respectively; heart rate was 87 beats/min; cardiac index (CI) was 4.42 liters/min/m2; systemic and pulmonary vascular resistances were 1931 and 194 dynes.sec.cm-5, respectively; oxygen delivery, consumption and extraction were 790 and 164 ml/min/m2 and 20.5%, respectively. This study represents a collation of cardiopulmonary values obtained from a large number of dogs (97) from a single laboratory using the same measurement techniques.     

Effect of hyperbaric oxygen on experimental acute distal colitis

It has been demonstrated that hyperbaric oxygen (HBO) is useful as an adjunctive therapy for Crohn's disease. However, its effects on ulcerative colitis have not been investigated. In the present study, HBO was tested for acetic acid-induced colitis, and antioxidant systems were evaluated to clarify its possible mode of action. Thirty-six Sprague-Dawley rats were randomly divided into three groups: sham control (Group I), colitis induced by acetic acid without any therapy (Group II), colitis induced by acetic acid and treated with HBO (Group III). HBO was given for 5 days, 2 sessions per day at 2.5-fold absolute atmosphere pressure (ATA) for a period of 90 min in rats in which colitis had been induced (Group III). Rats were sacrificed on the 5th day after the procedure. Superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH Px) activity were measured in the intestinal tissue and erythrocyte lysate. MDA and GSH Px were also determined in the plasma. Whereas MDA levels in erythrocyte, plasma and intestinal tissue were decreased, the levels of GSH Px and SOD were significantly increased in Group III as compared to those of Group II. The results of our study suggest that hyperbaric oxygen therapy has beneficial effects on the course of experimental distal colitis and that antioxidant systems may be involved in its mode of action.