Antioxidant parameters of horses according to age, sex, breed and environment.

Antioxidant defences interact to form an integrated system. There is no comprehensive and uniform view on issues concerning the antioxidant status in horses. Therefore, the aim of the present study was to estimate antioxidant parameters in horses of different age, sex and breed as well as environment and relationship between different antioxidants. Parameters of selected antioxidants including superoxide dismutase (SOD), glutathione peroxidase (GSH-px), total antioxidant status (TAS), ceruloplasmin (CP), bilirubin, uric acid, zinc, copper and selenium were determined in blood of 80 clinically healthy horses. Antioxidant parameters significantly varied between horses form different environments or different breed and sex. Age of horses had no significant effect on antioxidant parameters. Significant correlation coefficients were observed between enzymatic and nonenzimatic antioxidants. The nature of the relationship between the antioxidant system in horses with respect to environmental factors is rather complex and to date only a part of system is known.

The effect of persistent chlorinated hydrocarbons on the secretion of estradiol and progesterone by bovine granulosa cells in vitro.

The role of persistent chlorinated hydrocarbons (PCH) in the reproductive disorders in ruminants is not well documented. In the present study we have examined the effect of these compounds and their metabolites on the secretion of estradiol (E2) and progesterone (P4) by bovine granulosa cells in vitro. Granulosa cells were isolated from large follicles (> or = 8 mm diameter) by gently washing the internal follicle wall. Aliquots of approximately 4 X 10(5) viable granulosa cells in 0.5 ml medium were cultured at 37 degrees C in an atmosphere of 5% CO2 and 95% air. Granulosa cells were cultured for 96h in a medium containing different concentrations (10(-1)-10(-4) ng/ml) of a PCH combination. Estradiol and progesterone were measured in unextracted granulosa cell culture medium by Enzyme Immunoassay (EIA). The exposure of granulosa cells to a combination of these organochlorine compounds in vitro results in a slight decrease of estradiol secretion only at the highest studied concentration of the PCH combination. However, the secretion of progesterone by these cells was seriously decreased, even by concentrations found in ovaries from animals kept under natural environmental conditions. The in vitro culture system of granulosa cells from preovulatory follicles may be useful in screening toxic effects of pesticides in animal reproduction.

Evaluation of the effect of experimental cow endometritis on bactericidal capability of phagocytizing cells isolated from the blood and uterine lumen.

Studies were undertaken to assess the bactericidal activity of phagocytes isolated from blood and the uterine lumen of clinically healthy cows after ovulation, and from cows in which endometritis was induced experimentally. Experiments were carried out on 28 clinically healthy cows of the black and white lowland breed. Animals were aged 5 years and were used between the 2nd and 8th day after spontaneous ovulation. Cows were divided into four groups. Group I comprised animals in which cell-mediated type immune reaction was induced in the left uterine horn by intrauterine challenge with tuberculin. Cows in this group were initially vaccinated with M. bovis via the intrauterine route. In group II, Arthus type immune reaction was induced by challenging immunized animals with C. fetus ssp. veneralis through intrauterine instillation. The non-specific inflammatory process was initiated in the uterus of animals in group III by one instillation of lipopolysaccharide from S. abortus equi. Animals in group IV were set as control and received a phosphate buffered saline instillation into the uterine lumen. The cells from the left uterine horn were washed out 6 h after induction. Neutrophils were isolated from blood samples collected from all animals within the same time. The bacterial activity of cells from the uterine lumen and blood was assessed with the nitro-blue tetrazolium reduction test. Results are presented as increase in optical density resulting from a constant number of phagocytizing cells (delta OD/10(6) cells). Induction of cell-mediated immune reaction or Arthus type immune reaction in the uterus significantly boosts the intracellular capability of uterine cells to kill bacteria through the oxidation system. Experimentally induced non-specific endometritis weakens the bactericidal activity of uterine phagocytes, while peripheral blood phagocytes efficiently kill the engulfed bacteria.

In vitro assessment of the influence of different leucocyte count lavaged from normal and inflamed cow uterus on the mouse embryo development.

The aim of this study was to evaluate the effect of different leucocyte count from normal and inflamed uterus in excess of PMNs on the development of mouse embryos during the morula or blastocyst stages in vitro. Results showed that cultivation of blastocysts in the presence of leucocytes washed from the uterus, after mediation of specific or nonspecific endometritis at a concentration of 1 x 10(4), led to significant inhibition of embryo development. No similar observations were made with blastocysts cultivated in the presence of leucocytes obtained from the normal uterus at 2-4 days after spontaneous ovulation.

Phagocytic activity of polymorphonuclear leukocytes lavaged from the lungs of horses with clinically diagnosed chronic pulmonary disease.

The aim of this study was to compare phagocytic activity of polymorphonuclear cells (PMNs) from the bronchoalveolar lavage of clinically healthy horses and those with severe chronic bronchiolitis. Research was carried out on 28 horses. Chronic inflammation of the lower airways was diagnosed in nine horses. Cells from the respiratory tract were lavaged according to accepted methods. For comparison, PMNs were isolated from peripheral blood of all investigated horses. The phagocytic activity of PMNs was determined in relation to two standard strains of Staphylococcus aureus, Staph, aureus Smith which was phagocytized after previous opsonization, and Staph, aureus 305, phagocytized without opsonization. From the investigations, it is shown that the PMNs present in the terminal airways of horses with severe chronic bronchiolitis are characterized by decreased phagocytic activity in relation to opsonized Staphylococcus aureus Smith and increased activity in relation to non-opsonized Staphylococcus aureus 305, as compared to the PMNs lavaged from the terminal airways of clinically healthy horses. No changes in the phagocytic activity of the peripheral blood PMNs were observed between clinically diseased horses and healthy horses.

The effect of organic chlorine compounds and their metabolites present in human milk on newborn mice.

The effect of pesticides and their metabolites (DDE, DDT, DDD, alpha, beta and gamma-HCH and PCBs) isolated from human milk on the blood and liver morphology of the mouse were studied. Mouse neonates were fed an extract of the organochlorine compounds in linseed oil for a period of 6 weeks. The lowest dose used in the experiment equalled that which a human infant can receive with its mother's milk, calculated per gram of body weight. Doses 10 and 100 times higher were also used. At the end of the experiment, tissue samples for electron microscopy and blood samples for haemotological examination were taken. Haematological examinations, in mice receiving pesticides, revealed a significant rise in the number of Heinz bodies in erythrocytes and an increased number of lymphocytes. These changes were related to the concentrations of the organic chlorine compounds that the animals were given. Electronmicroscopy demonstrated that organochlorine pesticides at doses equal to that might be received by infants caused proliferation of the smooth endoplasmic reticulum in hepatocytes without any clear damage to other subcellular structures. Doses 10 and 100 times higher resulted in more extensive proliferation of the smooth endoplasmic reticulum, reduction of the rough elements of the endoplasmic reticulum and damage to mitochondria. The latest changes were associated with an increased number of prominent Kupffer cells and the appearance of immigratory cells with traits characteristic of lymphocytes and monocytes.

Effect of organochlorine pesticides on the phagocytic activity of bovine milk cells.

The effect of a complex of organochlorine compounds on the phagocytic activity of macrophages and neutrophils in milk was assessed in vitro using a mixture of DDT, DDE, DDD, alpha, beta and gamma-HCH and HCB and PCBs in concentrations occurring in milk and in concentrations being multiples of the concentrations in milk. Standard strains 305 and Smith of S. aureus were used for phagocytosis assessment. A suppressing effect of these compounds was observed on the per cent of cells phagocytizing both these strains, with a significant decrease in the number of bacteria of both strains phagocytized by macrophages, and the number of S. aureus strain 305 bacteria phagocytized by neutrophils. The concentrations of the mixture of organochlorine compounds used in the experiment had no significant effect on the number of Smith strain bacteria phagocytized by neutrophils.

Hydroxyurea-induced toxic side-effects in animals and an attempt at reducing them with vitamins E and C.

Using the changes in several blood parameters and in the histological picture of the liver as markers of toxicity, the effect of hydroxyurea (HU) in healthy rabbits or mice was examined during two weeks after a single administration of this drug. In rabbits a transient significant decrease in blood erythrocyte count with a gradual increase in their osmotic resistance, a suppression of granulocyte phagocytic capacity, and an elevation of acid phosphatase activity in the serum were found. An increase in the proportion of lymphocytes without detectable lysosomes as tested by supravital staining also appeared. No significant difference was observed in the white cell count and lipoperoxide levels after HU administration. Histological picture of the liver, excised from HU-treated mice, indicated a marked hepatotoxicity of the drug. Some of the toxic effects were reduced in animals supplemented with vitamins E and C.

Mechanism of unbalanced growth-induced cell damage. I. A probable role for hydrolytic enzymes synthesis.

This study examines the relationship between cell progress into the state of unbalanced growth, hydrolytic enzyme activities and cell survival during the exposure of L5178Y cells to hydroxyurea (HU), excess thymidine (dThR), hydroxyurea with excess of four deoxyribonucleosides (dNR) or excess dTHR with deoxycytidine (dCR). Cell progress into the state of unbalanced growth was measured as cell size, protein/DNA ratio and protein content per cell. Activities of two lysosomal (acid phosphatase, beta-N-acetylglucosaminidase) and one cytoplasmic non-lysosomal (LDH) enzymes were determined. It has been found that in cells arrested by HU or excess dThR, a progressive cell volume increase with protein/DNA imbalance is correlated with a progressive increase in lysosomal and non-lysosomal hydrolase activities in the cells and in the medium and with a marked lethal effect. Cell volume increase, enhancement of enzyme activities and cell killing could be prevented in HU-arrested cells by concomitant addition of excess dNR (deoxyadenosine, deoxyguanosine, thymidine, deoxycytidine) leading to equal inhibition of DNA and protein synthesis. Control-like values of all parameters were achieved also in cells in which the dThR-inhibiting effect was reversed by dCR addition. It is suggested that a common pathway in the mode of action of the chemotherapeutic agents inducing cell killing through the state of unbalanced growth can be the over-production, abnormal accumulation and progressive leakage of numerous hydrolytic enzymes through the cell membranes, leading in consequence to 'lytic' cell death.

Amelioration of hydroxyurea-induced suppression of phagocytosis in human granulocytes by free radical scavengers.

The exposure of human circulatory white cells in vitro to 0.1-1-10 mol/l hydroxyurea (HU) for 20 h induced a progressive dose-dependent suppression of the phagocytic activity of granulocytes. The suppressing effect of 20 h exposure to 1 mol/l HU was used to examine the protection afforded by free radical scavengers against HU-induced cytotoxicity. It has been found that, in the suitable concentration of the protecting agent, a substantial protective effect of sodium benzoate, acetylosalicylic acid, alpha-tocopherol, ascorbic acid, catalase, peroxidase or superoxide dismutase can be achieved.

Effect of a protein-free haemodialysate on tissue respiration and healing of burns.

Investigations were carried out in three groups of rats (10 rats per group) sustaining partial skin thickness burns covering 2.2 per cent of body surface area. Thirty minutes after burning animals received: Group I, 1 ml Solcoseryl, IP; Group II, 1 ml normal saline, IP; Group III, no treatment (controls). Two hours after burning tissue respiration and the tissue concentrations of potassium, magnesium, calcium and sodium ions were measured in biopsies from the burn wounds. It was found that early post-burn administration of Solcoseryl significantly inhibited the secondary destructive processes and accelerated the regeneration of damaged tissue.

Hydroxyurea has the capacity to induce damage to human erythrocytes which can be modified by radical scavengers.

The treatment of human erythrocytes with hydroxyurea [HU] results in the azide-dependent changes in osmotic fragility and in increased methemoglobin formation. Similar changes were induced by H2O2 treatment. However when H2O2 in the presence of azide stimulated malondialdehyde production, in the HU-treated cells no malondialdehyde was detectable. When subjected to an oxidant stress [sodium ascorbate] HU-treated erythrocytes were more fragile and revealed changes in the absorption spectrum of the TBA-reactive material in comparison with the cells treated with ascorbate alone. Partial protection by radical scavengers against certain HU-induced changes can be achieved. The results indicate that HU can damage erythrocytes and suggest the radical origin of these effects.