Marcin Zawadzki

Melatonin stimulates the activity of protective antioxidative enzymes in myocardial cells of rats in the course of doxorubicin intoxication.

Abstract: The study aimed at determining the effect of melatonin on the activity of protective antioxidative enzymes in the heart and of lipid peroxidation products in the course of intoxication with doxorubicin (DOX). The rats were categorized into four groups, receiving: 0.9% NaCl i.p. (NaCl control); melatonin [20 mg/kg body weight (b.w.)] s.c. (control Mel); DOX (2.5 mg/kg b.w.) i.p.; melatonin plus DOX in doses as above. All the substances were administered once in a week for four consecutive weeks. Homogenates of heart tissue were examined for activities of glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), levels of reduced glutathione (GSH) and of lipid peroxidation indices (MDA + 4‐HDA). Administration of melatonin alone did not induce alterations in levels of MDA + 4‐HDA, GSH, or in activity of GPx, SOD or CAT, as compared to the group receiving 0.9% NaCl. GSH levels decreased following DOX but remained at normal levels following DOX and melatonin. The level of MDA + 4‐HDA increased following DOX, as compared with the control, a change prevented by the combination of DOX+melatonin. Activities of GPx, SOD and CAT were higher in groups receiving DOX and/or DOX plus melatonin than in control groups. Activity of CAT and the level of GSH in the group receiving DOX plus melatonin were significantly higher than in the group intoxicated with DOX alone. The obtained results demonstrate that, when given in parallel with DOX, melatonin protects cardiomyocytes from damaging effects of the cytostatic drug (reflected by the levels of MDA + 4‐HDA). The protective effect resulted, in part from the augmented levels of GSH and from stimulation of CAT activity by melatonin in cardiomyocytes subjected to the action of DOX.

Influence of selenium and/or magnesium on alleviation alcohol induced oxidative stress in rats, normalization function of liver and changes in serum lipid parameters.

The aim of this study was to evaluate the attenuating effect of given selenium and/or magnesium on ethanol-induced oxidative stress, disturbances of liver function and cholesterol metabolism. Forty male rats were divided into five groups: C – control, Et – intoxicated with alcohol (15% solution in drinking water), Et + Mg, Et + Se, Et + Mg + Se – intoxicated with alcohol and supplemented with selenium (0.4 mg Se/l water), magnesium (100 mg Mg/l water) and combination of Se and Mg, respectively. The experiment was carried out over the 3 months. The results show that the chronic ingestion of alcohol induces lipid peroxidation and histopathological changes in liver. Supplementation with magnesium only partially alleviates oxidative stress and damages in this tissue. The both selenium alone and combination of magnesium and selenium significantly elevated total antioxidant status (TAS) in serum, activity of glutathione peroxidase and ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in liver and retarded oxidative stress and histopathological changes in this tissue. Chronic administration of ethanol (alone and with magnesium) resulted in significant decrease in the serum total cholesterol and retardation in the body weight gain in comparison with the control group. In the groups supplemented with selenium and selenium and magnesium simultaneously, concentration of total cholesterol in serum and body gains was similar to the control group. Supplementation of Se or selenium and magnesium simultaneously significantly enhances antioxidant defence and is more effective against alcohol-induced oxidative stress, disturbance of liver function and cholesterol metabolism than the separate use of magnesium.

Effects of melatonin on lipid peroxidation and antioxidative enzyme activities in the liver, kidneys and brain of rats administered with benzo(a)pyrene.

Benzo(a)pyrene [B(a)P] is a widespread pollutant with a mutagenic, carcinogenic and strong prooxidative properties. The present study evaluated the melatonin effects on lipid peroxidation products levels and on activity of antioxidative enzymes in the course of B(a)P intoxication. Control rats were treated with 0.9% NaCl; another group was given 10mg melatonin/kg bw; a third group was injected twice a week with B(a)P at the dose of 10mg/kg bw; the fourth group received both B(a)P and melatonin at the dose as mentioned above. The experiment continued for 3 months. In homogenates of brain, liver and kidneys lipid peroxidation was appraised by evaluation of malonyldialdehyde and 4-hydroxyalkenal (MDA+4HDA) levels. Activities of glutathione peroxidase (GPx), superoxide dysmutase (SOD) and catalase (CAT) and concentration of reduced glutathione (GSH) were also estimated. In animals receiving both B(a)P and melatonin, lower levels of MDA+4HDA were observed in all organs as compared to the group treated with B(a)P only. Following administration of B(a)P, GSH level decreased in brain and kidney. Melatonin in combination with B(a)P induced rises in the GSH level in liver and brain, as compared to the receiving B(a)P alone. The activity of SOD increased in the rats treated with melatonin alone but the highest activity was observed in rats treated with B(a)P plus melatonin. CAT activity in the melatonin-treated group increased in brain and liver. Similar to SOD, activity of the enzyme significantly increased in the group treated in combination with B(a)P and melatonin, as compared to the remaining groups in all tested tissues. The results suggest that melatonin protects cells from the damaging action of B(a)P. According to our knowledge, there are no studies describing the effects of melatonin on lipid peroxidation markers and antioxidative enzymes during intoxication of B(a)P in the brain, liver and kidneys. The results of present study give a perspective for further studies of its free radical scavenger properties in prevention of oxidative stress dependent diseases, among others cancers caused by carcinogens such as B(a)P.

Vipera berus bites in the Region of Southwest Poland--a clinical analysis of 26 cases.

OBJECTIVE Vipera berus is the only naturally occurring venomous snake in Poland. Its venom is primarily vasculotoxic and evokes both local and systemic findings. The aim of the study was to review a series of clinical cases of V berus bites occurring in southwest Poland. METHODS The charts of 26 patients (age range, 16-66 years; mean, 42 years) hospitalized with V berus bites were retrospectively analyzed using a data collection tool. Demographic and clinical data were extracted. RESULTS The most common local findings of envenomation were edema of the bitten limb with associated extravasations observed in 24 (92.3%) patients, but in only 1 (3.8%) case did the edema spread to the trunk. In 22 (84.6%) cases edema disappeared within 2 weeks after the bite. Systemic disturbances observed in the patients were: shock (1 case), mild transient hypotension (1 case), prolonged hypotension (3 cases), bronchospasm and laryngeal edema (1 case), diarrhea (1 case), transient supraventricular arrhythmias (2 cases), neutrophilic hyperleukocytosis (2 cases), and thrombocytopenia below 50000 cells/microL (5 cases). In 16 patients (61.5%) the envenomation was classified as moderate and this type was predominant. Six cases were classified as severe. No fatal case was reported. Treatment included the administration of specific antivenom in 14 cases (in all severe and half of moderate cases) and symptomatic treatment applied in all cases. CONCLUSIONS Moderate envenomation prevailed among the patients analyzed in the study. Antivenom treatment is primarily necessary in cases of severe (grade 3) and in some cases of moderate (grade 2) envenomation, especially in patients with persistent or recurring hypotension.

Suicidal overdose with relapsing clomipramine concentrations due to a large gastric pharmacobezoar

The paper presents a case of fatal intoxication after massive sustained-release clomipramine overdosage with prolonged toxicity related to a large gastric pharmacobezoar. 42-year-old female was admitted to the toxicology unit 14 h after drugs ingestion. At admission patient was deeply unconscious, required controlled mechanical ventilation. Serum total level of TCAs was 1955 ng/mL. Gastric lavage revealed no pills. Within the next 12h the patients clinical condition improved. TCAs level decreased to 999 ng/mL. However, after another 10h the clinical condition started deteriorating again and the patient went into a deep coma requiring controlled mechanical ventilation. TCAs level increased to 2011 ng/mL. X-ray and computed tomography revealed large pharmacobezoar consisted from radio-opaque pills. In the 28th h of hospitalization gastrotomy was performed, confirming presence of pharmacobezoar formed from Anafranil SR tablets. After surgery TCAs level was gradually decreasing. However, the patients condition did not improve, she died 32 h after gastrotomy. Post-mortem analyses revealed drug and its metabolite toxic levels in blood (clomipramine - 1729 ng/mL, norclomipramine - 431 ng/mL) and toxic levels in internal organs: myocardium (clomipramine - 14,420 ng/g, norclomipramine - 35,930 ng/g), vitreous humor (clomipramine - 1000 ng/mL, norclomipramine - 3110 ng/mL). Described case report indicates that sustained release clomipramine tablets may form pharmacobezoar. X-ray and computed tomography examinations should be considered in cases of massive abuse of sustained release clomipramine, particularly if symptoms of intoxication are recurrent or persistent.

The influence of road transport on the activities of glutathione reductase, glutathione peroxidase, and glutathione-S-transferase in equine erythrocytes.

BACKGROUND Transport of horses may have significant impact on serum biochemical and hematologic analytes and resistance to infection. OBJECTIVE The aim of our study was to assess the influence of transport stress on selected enzymatic antioxidants in equine blood. METHODS The study was conducted on a group of 60 horses of different breeds and ranging in age from 4 to 10 years. Venous blood was collected immediately before loading horses onto trailers for 8 hours of transport (I), immediately after unloading them from the trailer (II), and after subsequent stall rest for 24 hours (III). Hemolysates of blood were prepared, and hemoglobin (Hb) concentration and activities of the enzymatic antioxidants glutathione reductase (GR), glutathione-S-transferase (GST), and glutathione peroxidase (GPx) were measured. Enzyme activities were expressed as units of activity per gram of hemoglobin. RESULTS There were significant decreases in activities (mean ± SD U/g Hb [minimum-maximum]) of GPx between collection times I (36 ± 14 U/g Hb [9-67 U/g Hb]) and III (30 ± 11 U/g Hb [12-51 U/g Hb]) and of GR between collection times I (54 ± 28 U/g Hb [7-117 U/g Hb]) and II (40 ± 23 U/g Hb [12-145 U/g Hb]). There was no significant difference in activities of GR between collection times I and III (50 ± 27 U/g Hb [9-116 U/g Hb]). There were no differences detected in GST activity among the 3 collection times. CONCLUSION Road transport has an impact on activities of the antioxidant enzymes GPx and GR, with recovery of GR activity evident by 24 hours post-transport. Decreased activity of these enzymes may be one mechanism for increased susceptibility to infections that are manifest after shipping; alternatively, decreases may indicate utilization as these enzymes work to neutralize increases in reactive oxygen species.

Fatal intoxication with hydrocarbons in deltamethrin preparation.

Pyrethroid insecticides are very widely used in agriculture and household due to high effectiveness and low toxicity to humans. We have described a case of a fatal oral intoxication with decis, the insecticide containing pyrethroid (deltamethrin) in a hydrocarbon base. Pyrethroids, including deltamethrin, undergo rapid biotransformation by liver enzymes, which limit their systemic toxicity. Thus, we assume that in the presented case, fatal outcome of poisoning with decis was rather connected with toxic effects of hydrocarbon base (solvent naphtha) than with deltamethrin action. In the described case, detection of aromatic hydrocarbons in blood and lung tissue and their metabolites in urine confirms that these substances were absorbed from gastrointestinal tract to the systemic circulation. Predominant among the clinical outcomes in our patient was profound depression of CNS with apnea, which could be connected with narcotic action of organic solvents. The cardiac arrest was in mechanism of asystolia with prior non-responsive to catecholamines bradycardia and vascular collapse. We connect it with hydrocarbon-induced cardiotoxicity. It is worth remembering that many pyrethroid-containing insecticides are formulated in a hydrocarbon base. Intoxication with such preparations should always be considered not only as poisoning with pyrethroid alone but also as intoxication with hydrocarbons.

The influence of 9-day trekking in the Alps on the level of oxidative stress parameters and blood parameters in native lowlanders

BACKGROUND The stimuli acting on a person in a high mountain environment (such as hypobaric hypoxia with subsequent reoxygenation, physical exercise) can significantly increase oxidative stress, stimulate erythropoiesis, lead to changes in the blood count and participate in the development of altitude sickness. OBJECTIVE The aim was to investigate changes in haematological parameters, indicators of oxidative stress (malondialdehyde - MDA) and antioxidant defences: catalase (CAT), superoxide dismutase (SOD), and total antioxidant status (TAS) in the plasma of young, healthy people after a 9-day expedition in the Alps. MATERIALS AND METHOD A total of 5 patients (4 men and 1 woman), members of the Wrocław Mountaineering Club, aged 24-26 years. Collection of blood samples was carried out immediately before departure and 3 days after the end of exposure to high-altitude conditions. During the expedition, the subjects were exposed to heights: 2,050-4,165 m.a.s.l., and exercise associated with climbing. RESULTS Trekking in the Alps neither caused significant changes in the parameters of red blood cells nor increased the level of oxidative stress parameters in plasma. CAT activity increased, the ratio of SOD / CAT decreased. There was also a decrease in the total number of leukocytes, mainly monocytes and basophils. CONCLUSIONS 9-day exposure to high-altitude conditions is not a substantial burden for the organism of young, physically active people. The increase in antioxidant capacity is sufficient to stop oxidative processes, which are severe in these conditions, and to prevent the occurrence of significant oxidative stress. Discontinuation of exposure to allergens and dust pollution clears the airways, which is indicated by the reduction in the number of monocytes and basophils.

Serum biochemical reference intervals for the Polish Konik horse (Equus cabalus gmelini Ant.)

BACKGROUND Establishment of reference intervals (RI) for serum biochemical analytes is important for monitoring the health of different breeds of horses. OBJECTIVES The aim of this study was to measure and report RI for serum biochemical analytes of the Polish Konik horse. MATERIAL AND METHODS Blood samples were collected from 74 clinically healthy Polish Konik horses living under controlled natural conditions. These were adult primitive horses, aged 3-15 years, including 28 males (21 stallions, 7 geldings) and 46 mares. Serum analytes were measured and analyzed using a commercial automated analyzer. RESULTS The following RI (medians) were comparable to previously published RI in horses: albumin 34.8-41.3 g/L (38.0); ALP 124-309 U/L (216); ALT 6-33 U/L (19); AST 300-566 U/L (433); calcium 2.8-3.2 mmol/L (3.0); chloride 95-102 mmol/L (99); cholesterol 2.1-3.4 mmol/L (2.7); CK 167-488 U/L (328); creatinine 101-170 μmol/L (136); glucose 2.7-5.3 mmol/L (4.0); lipase 303-1143 U/L (723); magnesium 0.8-1.0 mmol/L (0.9); phosphate 0.9-1.5 mmol/L (1.2); potassium 2.4-5.0 mmol/L (3.7); sodium 138-144 mmol/L (141); total bilirubin 8-16 μmol/L (11.7); total protein 63.5-78.9 g/L (71.2); triglycerides 0.0-0.3 mmol/L (0.1); urea 2.2-7.3 mmol/L (4.7). CONCLUSION Reference intervals established in this study provide a valuable preliminary baseline for assessment of serum analytes in healthy Polish Konik horses.

Influence of 8-Hour Road Transportation on Selected Physiological Parameters in Horses

Abstract The objective of this study was to assess the physiological responses in mature healthy horses prior to an 8 h transport, as well as immediately and 24 h after transportation in a horse truck, under autumn conditions typical for central Poland. The study was conducted on a group of 60 horses of different breed. Horses ranged from 4 to 10 years of age. Venous blood was taken 3 times: I - directly before loading the horses to the trailer, II - directly after unloading the horses from the trailer, III - after period of 24 h resting. There was no change in the concentrations of urea, cholesterol, creatinine, and magnesium and in the activity of alkaline phosphatase during the three subsequent samplings. Albumin, total protein, alanine aminotransferase, aspartate aminotransferase, total bilirubin, creatine kinase, and triglycerides were mostly influenced by the transport. The obtained results indicate that the transportation of horses alters concentrations of physiological variables of their metabolism.