Estimation In Vivo of the Antioxidant Activity of Axillary-Blueberry (Vaccinium axillare Nakai) Fruit under Oxidative Stress.

The intensity of free radical oxidation processes in vivo (a model of induced oxidative stress) was studied after a probe administration of the fruit extract of the axillary blueberry (Vaccinium axillare Nakai). \Four groups (n = 40) of white CBA line male mice weighing 20-25 g were involved in the experiment: (1) intact control; (2) introduction of a 0.9% sodium-chloride solution orally for 10 days, a dose of 10 mL/kg per day; (3) "cisplatin" group (animals received 0.9% sodium-chloride solution similarly to group 2; on the fifth day of the experiment, cisplatin was administered one time by intraperitoneal injection at a dose of 7.5 mg/kg); and (4) "cisplatin + blueberry" group (mice received orally an extract of axillary-blueberry fruits at a dose of 10 mL/kg per day for 10 days; on the fifth day of the experiment, cisplatin was administered one time by intraperitoneal injection at a dose of 7.5 mg/kg). The antioxidant activity of the axillary blueberry was studied by a method of chemiluminescence. The analysis of kinetic parameters of chemiluminescence of mouse-kidney homogenate demonstrated that, against the background of a single intraperitoneal injection of cisplatin, oxidative stress develops in animals, with its severity decreasing under the action of axillary blueberry-fruit extract. The axillary blueberry-fruit extract has pronounced antioxidant properties and can play a role in the treatment and prevention of disease associated with oxidative stress.

Role of Amino Acid Arginine and Nitric Oxide in Mechanisms of Cytoprotective Effect of Non-Opiate Leu-Enkephalin Analogue In Vitro.

Incubation of primary culture of pulmonary fibroblasts with non-opiate analogue of leuenkephalin (NALE; Phe-D-Ala-Gly-Phe-Leu-Arg, 0.1 µM) reduced generation of superoxide anion-radical (by 20.7%) and decreased the number of p53+ cells (by 40.2%) induced by exposure to H2O2 (60 µM). The cytoprotective effect of NALE was potentiated by NO synthase inhibitor L-NAME (1 mM): the number of p53+ cells decreased by 65.3% and morphometric parameters of the cell nuclei and nucleoli were improved. Incubation of pulmonary fibroblasts culture with peptide G (Phe-D-Ala-Gly-Phe-Leu-Gly, 0.1 µM) also significantly reduced the damaging effect of H2O2: the number of p53+ cells decreased by 73.5%, the area of cell nuclei returned to normal, and generation of superoxide anion-radical decreased by 18.4%. These results indicate that C-terminal amino acid Arg and activation of NO synthase are not involved in the direct cytoprotective effect of NALE.

Hepatoprotective Effect of Neonatal Administration of Non-Opioid Leu-Enkephalin Analogue in Adult Albino Rats Subjected to Antenatal Hypoxia.

We studied the effects of neonatal administration of non-opioid leu-enkephalin analogue peptide NALE) on the morphological parameters of the liver and redox status of 60-dayold albino rats subjected to antenatal hypoxia. In animals subjected to antenatal hypoxia, a decrease body weight and reduction of the area of hepatocytes and total area of their nucleoli were observed; these changes were accompanied by activation of free-radical processes and impairment of antioxidant defense in the liver and blood serum. Intraperitoneal administration of NALE (100 µg/kg, daily) during postnatal days 2-6 normalized body weight, hepatocyte area, and total area of their nucleoli, significantly reduced the intensity of ROS generation, and improved antioxidant protection in the liver and blood serum in 60-day-old animals subjected to antenatal hypoxia.

Immediate and Delayed Effects of Food Restriction on Some Parameters of Brain Development in Rats.

The effects of 15-day restriction diet (from 1 to 1.5 months of age) on some parameters of brain development were studied in rats. The immediate and delayed (15 days after transfer to normal ration) effects were evaluated. The immediate effects included a significant decrease of the absolute weights of the brain and hemispheres. The relative weight of the brain was significantly higher. The thickness of the cortex of the parietal lobe proper and its layer I decreased. The absolute weights of the brain and hemispheres were less than in the control 15 days after the rats were transferred to ad libitum feeding, while the relative weight of the brain was higher than in controls. The thickness of the parietal and anterior parietal cortex and the numerical density of neurons in layers II and V did not differ from the control. In the neurons of layers II and V of the anterior parietal and parietal lobe proper the nuclei were larger, while the nucleoli were enlarged in the neurons of these locations and the hippocampus, the shifts being significant in the anterior parietal layer V. The concentrations of RNA in the parietal, anterior parietal, and hippocampal lobe neurons in different groups were similar directly and 15 days after the diet. Changes in the gravimetric and morphometric parameters of the brain were paralleled by the development of oxidative stress.

Delayed Effects of Neonatal Administration of Non-Opioid Analog of Leu-Enkephalin on Cerebral Consequences of Antenal Hypoxia.

In 60-day-old male rats after antenatal hypoxia, the body weight and the absolute weight of the cerebral hemispheres were significantly lower than in the progeny of intact animals. Analysis of brain sections stained with silver nitrate revealed reduced number of nucleoli in neocortical layer II and hippocampal CA1 neurons and smaller area of neuronal nuclei in neocortical layer V and total area of nucleoli in neurons of all studied zones. The animals demonstrated increased locomotor activity in the elevated plus-maze test. Chemiluminescent analysis of brain homogenates revealed the presence of oxidative stress at the organ level. Neonatal administration of non-opioid analog of leu-enkephalin (peptide NALE) after antenatal hypoxia normalized body weight, neutralized morphometric changes in the nucleoli and nuclei of neurons in the neocortex and hippocampus, and improved oxidative status of the brain. In 30- and 60-day-old male rats subjected to antenatal hypoxia and receiving peptide NALE during the neonatal period, behavioral responses were partially normalized. Non-opioid analog of leu-enkephalin can be a promising drug for correction of cerebral consequences of antenatal hypoxia.

Correction of Negative Effect of Antenatal Hypoxia on Liver Tissue Homeostasis in Newborn Albino Rats with Opioid Peptides.

We studied the possibility of correction of the negative effects of antenatal hypoxia on the liver tissue homeostasis in 7-day-old albino rats by administration of opioid peptides in a dose of 100 µg/kg on postnatal days 2-6. Administration of mixed µ/δ-opioid receptor agonist Dalargin neutralized deviations of gravimetric indicators, parameters of proliferative activity, and activity of the nucleolar apparatus of hepatocytes. Administration of the non-opiate Leu-enkephalin analogue did not normalize gravimetric parameters and nucleolar apparatus parameters, however, it significantly increased the pool of proliferating hepatocytes. Both peptides significantly reduced the intensity of free radical oxidation, improved antioxidant antiradical defense and resistance to peroxidation in the liver tissue of animals subjected to antenatal hypoxia.

Cytoprotective Effect of Peptide Sedatin, an Agonist of µ/δ-Opioid Receptors, on Primary Culture of Pulmonary Fibroblasts of Albino Rats under Conditions of Oxidative Stress.

We studied the effects of a synthetic analogue of dermorphin peptide sedatin on DNA synthesis, nucleolar apparatus, and parameters of free radical oxidation in the primary culture of pulmonary fibroblasts under conditions of oxidative stress. Oxidative stress significantly enhanced production of superoxide anion radical in the culture, sufficiently inhibited DNA synthesis in fibroblasts, and reduced the size of cell nuclei and parameters of the nucleolar apparatus. Sedatin prevented accumulation of free radical oxidation products and changes in karyometry parameters induced by oxidative stress. The peptide completely eliminated changes in the parameters of fibroblast nucleolar apparatus and abolished the inhibitory effect of oxidative stress on the number of DNA-synthesizing cells. Pretreatment with non-selective opioid receptor antagonist naloxone hydrochloride partially abolished the effects of sedatin in the primary culture of pulmonary fibroblasts.

Effect of Proline-Containing Oligopeptides PGP and RGP on Proliferative and Protein-Synthesizing Activity of Cultured Pulmonary Fibroblasts under Conditions of Oxidative Stress.

We studied the effect of glyprolines Pro-Gly-Pro (PGP) and Arg-Gly-Pro (RGP) on the primary culture of pulmonary fibroblasts from newborn albino rats under normal conditions and during oxidative stress. Under physiological conditions, the peptides had no effect on parameters of cell division. Hydrogen peroxide induced intensive oxidative stress accompanied by suppression of protein-synthesizing function. When hydrogen peroxide was added to the culture containing the test peptides, correction of the oxidative status was observed accompanied by activation of DNA-synthesizing activity and inhibition of lucigenin-dependent chemiluminescence.

Formation of Hyaline Membranes in the Lungs of Albino Rat as a Result of Exposure to Bleomycin During the Neonatal Period.

We studied the effect of bleomycin (1 mg/kg) in 1-day-old rat pups (single intraperitoneal injection) and 4-day-old pups (3 injections on postnatal days 1, 2, and 3). The formation of hyaline membranes against the background of oxidative stress was documented in all bleomycin-treated rats. Repeated injection of bleomycin (4-day-old pups) led to more pronounced destructive alterations in the lungs characterized by the formation of microcysts and atelectases as well as hypertrophy of the interstitial connective tissue. These alterations persisted in 14-day-old rats. Administration of bleomycin during the neonatal period can be a promising approach to simulation of hyaline membrane disease (respiratory distress syndrome).

Effects of Glyprolines on DNA Synthesis and Free Radical Oxidation in Mouse Gastric Mucosa Under Physiological Conditions and During Therapy with Oral Non-Steroid Anti-Inflammatory Drugs.

Studies by (3)H-thymidin autoradiography showed that injections of Pro-Gly-Pro and Arg-Gly-Pro peptides caused no changes in the DNA synthesis processes in the gastric mucosa. Both peptides induced a reduction of free radical oxidation activity, which was shown by chemiluminescence. Indomethacin induced lesions in the gastric mucosa, triggered oxidative stress, and reduced proliferative activity. Injection of Pro-Gly-Pro peptide before indomethacin corrected disorders in oxidative status and normalized DNA synthesis. Preinjection of Arg-Gly-Pro led to enlargement (by 4.6 times) of the focus of lesions in animals treated by indomethacin and augmented oxidative stress.

Effect of Antioxidant Echinochrome A on Bleomycin-Induced Pulmonary Fibrosis.

We studied the protective effect of antioxidant echinochrome A on bleomycin-induced pulmonary fibrosis in rats at the early stages of postnatal ontogeny. Administration of echinochrome A was shown to reduce the severity of bleomycin-induced oxidative stress in the lungs, prevented the development of hypertrophy of interalveolar connective tissue and peribronchial lymphoid infiltration, and normalized the ratio of volume densities of interalveolar septa and alveolar lumen.

Effects of antenatal hypoxia on tissue homeostasis in the myocardium of albino rats: early and delayed consequences.

Body weight, absolute and relative heart weight, DNA-synthetic activity of cardiomyocytes, and number of nucleoli in cardiomyocyte nuclei were reduced in newborn albino rats exposed to antennal hypoxia. All these changes developed against the background of oxidative stress. In mature animals, the absolute and relative heart weights were decreased, but the ratio of mononuclear cardiomyocytes in both ventricles was higher than in the control. These changes were accompanied by an increase in the percent of collagen fibers in the myocardial stroma. In the right ventricle, an increase in the cardiomyocyte length to width ratio and a decrease in the number of nucleoli were found. The observed changes can probably induce heart function disturbances under pathological conditions.

Effects of antenatal hypoxia on the parameters of tissue homeostasis in the liver of albino rats.

We studied age-dependent dynamics of some liver parameters in albino rats after antenatal hypoxia. A decrease in body weight, absolute liver weight in newborn and mature rats, and a reduction of relative weight of the liver in mature animals were observed under these conditions. It was found that antenatal hypoxia promotes a decrease in DNA-synthetic activity of hepatocytes, intensifies free radical processes in the liver of newborn rats, and reduces the number of nucleoli in hepatocytes. In mature animals, antenatal hypoxia was followed by a decrease in hepatocyte sizes and area of nucleoli, an increase in the number of binuclear cells relative to mononuclear cells, and a decrease in the number of nucleoli in hepatocyte nuclei. These data can reflect impairment of tissue homeostasis in the liver of mammals exposed to antenatal hypoxia.

Proliferation and local manifestations of oxidative stress in mucous membrane of the ileum in patients at early stages of hemorrhagic fever with renal syndrome.

Activation of proliferative processes (Ki-67 proliferation index) and local oxidative stress was found in biopsy material from the mucosa of the large intestine of patients at the early stages of hemorrhagic fever with renal syndrome on days 5-10 after disease onset. Analysis of data of rectosigmoidoscopy and clinical examination refl ected the presence of colitis. It can be suggested that activation of proliferation is typically associated with local oxidative stress during various pathological processes.

Effect of treatment with testosterone derivatives on morphometric characteristics and free radical oxidation in rat cerebral cortex.

Morphometric characteristics and free radical oxidation in neurons of the neocortex and hippocampus of male and female rats were studied 1 month after administration of Sustanon-250 in a dose of 8 mg/kg during the pubertal period of ontogeny. The weight of the brain was shown to decrease in males. In both female and male rats, the width of the anterior parietal lobe and the numerical density of neocortical and hippocampal neurons decreased. Sex differences were found in free radical oxidation in the cerebral cortex. The intensity of this process increased significantly in females, but decreased in males.

Acute cholestatic liver injury caused by polyhexamethyleneguanidine hydrochloride admixed to ethyl alcohol.

INTRODUCTION: Polyhexamethyleneguanidine hydrochloride (PHMG) is an antimicrobial biocide of the guanidine family. In the period from August 2006 to May 2007, more than 12500 patients were admitted to hospital with a history of drinking illegal cheap "vodka" in 44 different regions in Russia, of whom 9.4% died. In reality, the "vodka" was an antiseptic liquid composed of ethanol (≈93%), diethyl phthalate, and 0.1-0.14% PHMG (brand name "Extrasept-1"). MATERIAL AND METHODS: We performed an analysis of the clinical features and outcome in four poisoning treatment centers in the cities of Perm, Ekaterinburg, Irkutsk, and Khabarovsk. A total of 579 patients (215 females and 364 males) with similar symptoms were included. RESULTS: The main symptoms on admission included jaundice (99.7%), skin itch (78.4%), weakness (96%), anorexia (65.8%), dizziness (65.3%), nausea (54.8%), vomiting (22.6%), stomach ache (52.7%), diarrhea (32%), and fever (50%). Mild symptoms were found in 2.5% of cases, moderate in 63%, and severe in 34.5%. Laboratory results were (mean ± SD): total bilirubin 249 ± 158 µmol/L, direct bilirubin 166 ± 97 µmol/L, cholesterol 14 ± 8 mmol/L, alanine aminotransferase 207 ± 174 IU/L, aspartate aminotransferase 174 ± 230 IU/L, alkaline phosphatase 742 ± 751 IU/L, and gamma-glutamyltranspeptidase 1199 ± 1095 IU/L. Patients generally recovered over a period of 1-5 months, although high levels of alkaline phosphatase and gamma-glutamyltranspeptidase were still found in all patients examined after 6 months. Sixty-one patients (10.5%) died between 23 and 150 days after poisoning. Local cholestasis, inflammatory infiltration, and fibrosis developing into cirrhosis were found by liver biopsy. CONCLUSION: Acute liver injury caused by PHMG-hydrochloride or PHMG in combination with either ethanol or diethyl phthalate can be characterized as cholestatic hepatitis with a severe inflammatory component causing high mortality.

[Influence of occupational factors on free-radical process parameters in workers of Far East Railway].

Workers of Far East Railway were diagnosed as having various disorders of free-radical oxidation processes due to occupational factors. Oxidative stress parameters were changed even at preclinical stages of broncho-pulmonary diseases--that evidence could be useful for early diagnosis of pulmonary diseases in individuals exposed to hazards.

[Role of smoking in chronic broncho-pulmonary diseases in workers of Far East Railway].

Smoking contributes more than occupational factors into broncho-pulmonary diseases development and into free-radical process disorders among Far East Railway workers. The data obtained should be considered in specifying treatment and prophylactic measures in these workers.

Status of free-radical oxidation and proliferation processes in patients with atopic dermatitis and lichen planus.

We studied the status of proliferation processes (by Ki-67 expression) and biogenesis of free radicals (by chemiluminescence method) in skin biopsy specimens from patients with atopic dermatitis and lichen planus. The index of Ki-67-positive nuclei significantly (p<0.05) increased to 28.40±2.00% in patients with dermatitis and to 32.6±1.9% in patients with lichen planus vs. 8.65±1.31% in the reference sample. Decompensated accumulation of free-radical oxidation products and inhibition of detoxification systems in skin biopsy specimens indicate the development of local oxidative stress. After therapy consisting of two 10-day courses of thymodepressin injections (0.1%, 1.0 ml) over 30 days, normalization of epidermis proliferation was observed. Labeling index in atopic dermatitis and lichen planus significantly decreased (p<0.05) to 17.00±1.87 and 10.9±1.1%, respectively. The role of free radicals in the development of hyperregeneratory processes during dermatoses is discussed.

Gastroprotective effect of dalargin in gastropathy due to treatment with nonsteroid antiinflammatory drugs.

We studied the effect of dalargin and its analogue [Dala]2-leu-enkephalin on the gastric mucosa in indomethacin-receiving animals. Indomethacin treatment was followed by severe injury to the gastric mucosa, decrease in proliferative activity of the epithelium, and stimulation of free-radical processes in gastric tissues. Dalargin significantly decreased the area of erosive and ulcerative lesions, had a normalizing effect on proliferation of epithelial cells, and reduced the degree of oxidative stress. Administration of [Dala]2-leu-enkephalin did not improve the state of the gastric mucosa.