Morphological and molecular-biological features of glioblastoma progression in tolerant and susceptible to hypoxia Wistar rats.

Hypoxia is a major pathogenetic factor in many cancers. Individual resistance to suboptimal oxygen availability is subject to broad variation and its possible role in tumorigenesis remains underexplored. This study aimed at specific characterization of glioblastoma progression in male tolerant and susceptible to hypoxia Wistar rats. Hypoxia resistance was assessed by gasping time measurement in an 11,500 m altitude-equivalent hypobaric decompression chamber. Based on the outcome, the animals were assigned to three groups termed 'tolerant to hypoxia' (n = 13), 'normal', and 'susceptible to hypoxia' (n = 24). The 'normal' group was excluded from subsequent experiments. One month later, the animals underwent inoculation with rat glioblastoma 101.8 followed by monitoring of survival, body weight dynamics and neurological symptoms. The animals were sacrificed on post-inoculation days 11 (subgroup 1) and 15 (subgroup 2). Relative vessels number, necrosis areas and Ki-67 index were assessed microscopically; tumor volumes were determined by 3D reconstruction from histological images; serum levels of HIF-1α, IL-1ß, and TNFα were determined by ELISA. None of the tolerant to hypoxia animals died of the disease during observation period, cf. 85% survival on day 11 and 55% survival on day 15 in the susceptible group. On day 11, proliferative activity of the tumors in the tolerant animals was higher compared with the susceptible group. On day 15, proliferative activity, necrosis area and volume of the tumors in the tolerant to hypoxia animals were higher compared with the susceptible group. ELISA revealed no dynamics in TNFα levels, elevated levels of IL-1ß in the susceptible animals on day 15 in comparison with day 11 and tolerant ones. Moreover, there were elevated levels of HIF-1α in the tolerant animals on day 15 in comparison with day 11. Thus, the proliferative activity of glioblastoma cells and the content of HIF-1α were higher in tolerant to hypoxia rats, but the mortality associated with the tumor process and IL-1ß level in them were lower than in susceptible animals. Specific features of glioblastoma 101.8 progression in tolerant and susceptible to hypoxia rats, including survival, tumor growth rates and IL-1ß level, can become the basis of new personalized approaches for cancer diseases treatment in accordance to individual hypoxia resistance.

Morphofunctional Changes in Colon after Cold Stress in Male C57BL/6 Mice Susceptible and Tolerant to Hypoxia.

There are individual differences in the tolerance to hypoxia and stress. Stress can contribute to the development of various diseases, including inflammatory bowel disease. It was found that inflammatory bowel diseases in animals susceptible to hypoxia runs more severe course than in tolerant animals. We studied morphofunctional changes in the colon under conditions of modeled cold stress in male C57BL/6 mice susceptible and tolerant to hypoxia. The animals were daily subjected to cold stress (20 min at -20°C) for 2 weeks. Cold stress was followed by an increase in the volume fraction of goblet cells in the colon and production of mucins by these cells in mice tolerant to hypoxia and an increase in cell content in the lamina propria of the colon mucous membrane in animals susceptible to hypoxia. The number of serotoninproducing endocrine cells increased in both groups, but these changes were more pronounced in mice susceptible to hypoxia.

Morphological and Functional Peculiarities of the Immune System of Male and Female Rats with Different Hypoxic Resistance.

The morphological and functional peculiarities of the immune system are studied in adult male and female Wistar rats with high and low hypoxic resistance. Sex-specific differences in the subpopulation composition of the peripheral blood lymphocytes, not depending on the hypoxic resistance of animals, are detected: the males have lower absolute counts of T helpers and higher percentage of regulatory T cells than the females in the diestrus phase. Comparison of the morphofunctional status of the immune system in male and female (diestrus) rats with high resistance to hypoxia has shown a better developed subcapsular zone of the thymus, higher levels of anti-inflammatory cytokine TGF-ß, and lower absolute counts of cytotoxic T lymphocytes in the peripheral blood in the males. Males with low hypoxic resistance have higher counts of phase II thymic bodies in comparison with low-resistant females. Hence, morphofunctional differences in the immune system of male and female rats with different hypoxic resistance are detected, which can determine the course of inflammatory diseases.

Structural Changes in the Nervous Fibers of the Colon Mucosa in Experimental Acute Colitis.

Morphological, morphometric, and ultrastructural analysis of the nerve fibers in the colon mucosa was performed in C57BL/6 mice at various terms of development of acute colitis induced by dextran sodium sulphate. The nerve fibers were labeled with antibodies to pan-neuronal marker ßIII-tubulin. The progression of inflammatory and ulcerative processes in the mucosa on days 3-5 was associated with hyperplasia and hypertrophy of nerve fibers that peaked on day 7 after colitis induction. Ultrastructural analysis at all terms of colitis development showed moderate degeneration of axons. Thus, hypertrophy and hyperplasia of the nervous fibers in colon mucosa in experimental acute colitis correlated with aggravation of the ulcerative process in the mucosa. These changes are determined by alteration of histoarchitectonics and regenerative processes in the mucosa.

Morphological and Biochemical Characteristics of Prostate Hyperplasia during Sulpiride Treatment.

We studied morphological changes in the prostate ventral lobe, proliferative activity of the epithelium in prostate acini, and the levels of prolactin and prostate-specific antigen in the blood serum of Sprague-Dawley rats after repeated injections of sulpiride in a dose of 40 mg/ kg over 30 and 60 days and in 10 and 30 days after withdrawal. Morphological and morphometrical analysis of hyperplastic changes in the prostate ventral lobe was performed. Ki-67+ proliferating epithelial cells in the acini were counted. The dynamics of serum concentrations of prolactin and prostate-specific antigen was evaluated by ELISA. Morphological and morphometrical analysis and evaluation of the content of Ki-67+ cells demonstrated epithelium hyperplasia in the prostate ventral lobe after sulpiride treatment for 30 or 60 days and in 10 days after withdrawal, but serum level of prostate-specific antigen did not differ from the control. After 60-day sulpiride treatment and in 30 days after withdrawal, pronounced hyperplastic changes of prostate and elevated concentrations of prostate-specific antigen (but not prolactin) were observed. Thus, administration of sulpiride (40 mg/kg) to Sprague-Dawley rats for 60 days allows, by morphological criteria and serum level of prostate-specific antigen, to model stable hyperplastic changes in the prostate corresponding to benign prostatic hyperplasia in humans.

Morphological Characteristic of Melanoma B16 Progression in C57BL/6 Mice with High and Low Resistance to Hypoxia.

The features of B16 melanoma progression in male C57BL/6 mice with initially high and low resistance to hypoxia were studied. To assess the resistance to hypoxia, the mice were placed in a low-pressure chamber at a simulated altitude of 10,000 m. One month after testing, B16 melanoma was inoculated to high- and low-resistant animals. In 19 days after melanoma transplantation, the severity of melanoma progression was assessed by morphological and immunofluorescent methods. The expression of vegf-a and hif-1a in the liver of melanomabearing and control mice was evaluated by real-time PCR. Tumor growth progression was more pronounced in low-resistant mice, which was seen from high weight of the primary tumor node, relative necrosis area, proliferation rates (mitotic index and number of Ki-67+ cells), and expression of vegf-a gene in the liver. In high-resistant to hypoxia animals, the number of caspase-3+ cells dying by apoptosis was higher. The data on more rapid melanoma progression in mice with low resistance to hypoxia should be considered during the search of new prognostic markers and methods for therapy of malignant neoplasms.

[Vaccination with virus-like particles containing hemagglutinin protects the lungs of mice with postifluenza bacterial pneumonia: virological, microbiological and clinical data].

INTRODUCTION: Influenza is a severe viral disease, a frequent complication of which is a secondary bacterial pneumonia. Influenza vaccines prevent secondary bacterial complications. Virus-like particles are one of the promising areas for the development of new vaccines. The aim of this work is to study the correlation of the pathomorphological characteristics of the lungs with clinical, virological, and microbiological markers of the disease at vaccination with virus-like particles (VLPs), containing hemagglutinin (HA) of influenza virus (HA-Gag-VLPs) in a murine model of secondary bacterial pneumonia induced by S. pneumoniae after influenza infection. MATERIAL AND METHODS: BALB/c mice were vaccinated with VLPs containing influenza HA. After 21 days, mice were infected with two strains of influenza viruses, homologous and non-homologous, and 5 days after viral infection, were infected with S. pneumoniae. The vaccination effect was evaluated by morphological, virological (titer of the virus in the lungs) and microbiological (titer of bacteria in the lungs) data, and was confirmed by clinical data (survival, change in body weight). RESULTS: Immunization with HA-Gag-VLPs, followed by infection with a homologous influenza virus and S. pneumoniae, reduced the area of foci of inflammation, inhibited the replication of the virus and bacteria in the lungs, and also protected animals from death and reduced their weight loss. Immunization with HA-Gag-VLPs upon infection with a heterologous strain and S. pneumoniae did not affect these criteria. CONCLUSION: The immunization with HA-Gag-VLPs prevented the viral replication, providing a reduction of S. pneumoniae titer and the degree of lung damage, protecting animals from the disease in a murine model of secondary bacterial pneumonia, induced by S. pneumoniae, after influenza infection with homologous strain of the virus.

Ultradian Rhythms of Body Temperatures in Male Wistar Rats Maintained under Conditions of Constant Illumination.

We studied the dynamics of the body temperature in mature male Wistar rats maintained under condition of constant illumination. It was shown that body temperature under these conditions varied with a 3-5-h period. The daily dynamics of body temperature summed up over 20-23-day intervals showed a 4-h ultradian rhythm with maxima at 3.35-4.30, 7.35-8.30, 11.35-12.30, 15.35-16.30, 19.35-20.30, and 23.35-00.30 h. During these hours, pronounced (>0.9°C) increase in body temperature was observed by 1.6 times more often than in other eriods. Thus, there are periods during the day when the increase in body temperature in rats in the absence of light cues occurs more often than in other periods of the day. Hence, about 4-h ultradian rhythm of body temperature has an external synchronizer that differs from lighting conditions.

Infradian Rhythms of Resistance to a Dissociative Anesthetic in Wistar Male Rats under Normal Conditions and After Surgical Removal of the Adrenal Glands and Testes.

Daily dynamics of changes in the latency of a response to dissociative anesthetic tiletamine (time from injection to ataxia) was studied in mature Wistar rats. Both intramuscular and intravenous administration of the anesthetic was associated with 4-day oscillations of the latent period synchronous with the dynamics of changes in the concentration of glucocorticoid hormones. The period and phases of the infradian rhythm of resistance to the anesthetic remained unchanged after removal of both adrenal glands and testes and administration of corticosterone synthesis blocker trilostane diminishing the 4-day cycle of changes in corticosterone level. Therefore, hormones of the adrenal glands and testes do not play the key role in the mechanisms of formation of the 4-day infradian rhythm.

[Esophageal wall structure in people of elderly and senile age].

Using histological methods, the esophageal wall structure and the cytoarchitectonics of mucous membrane were studied in the individuals of elderly (n = 5) and senile (n = 10) age. The control group included the individuals of I (n = 3) and II (n = 3) periods of mature age. It was demonstrated that with advancing age in most cases the destructive processes took place in the epithelium (delamination of the layer, separation of large fragments, formation of microerosions etc.) in most of the studied cases. Lymphocytes, neutrophils and eosinophils were found between the epithelial cells; the numbers of infiltrating cells was increased 2-3 times during aging. Mucosal lamina propria and the submucosa, in particular, were characterized by the thickening of the bundles of collagen fibers. A two-fold increase in the number of the cells of the fibroblast lineage was found. The number of leukocytes in the lamina propria was increased by the eldery age in the upper and lower parts of the esophagus (3.5 and 1.75 times respectively). The changes in lamina muscularis were manifested by its thinning, delamination and myocyte dissociation. Remodeling of the muscular tunic was less pronounced. The degree of changes increased distally and varied widely depending on the individual peculiarities.

[Determination of genome dose of active ribosome genes and several quantitative parameters of extracellular dna in test-subjects of the experiment with 7-day immersion].

Genome dose of active ribosome genes (ARG), average of nucleolus argyrophil structures in lymphocyte nuclei, levels of extracellular DNA (DNA(e)) concentrations and ratio of antibodies to total DNA (AB(DNA)) and ribosomal DNA (AB(DNA-rib)), and nuclease activity were determined in peripheral blood of 8 volunteered subjects (21-26 y.o.) in the experiment with 7-d DI. Results of the investigation revealed a broad individual variability ensued from heterogeneity of the group of the test-subjects as to ARG values. There was an inverse negative relationship between ARG values and increment of the ribosome genes activity index. Part of the subjected exhibited increased DNA(e) levels on completion of the experiment, whereas the others decreased the parameter demonstrating individual character of body reaction. No correlation was established between DNA(e) content and nuclease activity in blood. Concentrations of AB(DNA) and DNA AB(DNA-rib) before and after immersion were essentially unchanged; however, they were higher as compared with the control group of blood-donors. Diversity of subjects' reactions was accounted to the broad range of ARG values. Therefore, selection of test-subjects for ground-based simulation experiments should be conducted with due consideration of the parameter.

[Morphological characteristics of the testicles of male Wistar rats intoxicated with morphine].

Experimental study of the effect of morphine on reproductive organs was made in Wistar rats. In the experimental animals from Group 1, 1% morphine solution was intraperitoneally injected in increasing doses during 10 days while in Group 2 animals were taught to administer morphine themselves during 28 days. Morphological and morphometric studies were conducted. The plasma levels of follicle-stimulating hormone, luteinizing hormone, corticosterone, testosterone, and estradiol were measured by enzyme immunoassay. It was ascertained that there was a progressive inhibition of both hormonopoietic and spermatogenic functions depending on the duration of morphine use.

[Morphofunctional changes in the pituitary-adrenal-gonadal system of Wistar rats exposed long to ethanol at free options].

Experiments were carried out on 40 adult male Wistar rats weighing 220-240 g, which used 15% ethanol or drinking water at free options during 2 months. Ten intact males using the drinking water ad libitum served as controls. Morphofunctional activity was morphologically and morphometrically studied in the anterior pituitary, adrenals, and testicles. The spermogram of epididymal sperm was examined. The plasma levels of follicle-stimulating hormone, luteinizing hormone, corticosterone, testosterone, and estradiol were measured by enzyme immunoassay. The rats using 15% ethanol for 2 months were observed to have feebly marked signs of the suppressed pituitary-adrenal-gonadal system depending on the dose of used ethanol. It is suggested that the amount of ethanol ingested by the animals depends on the baseline level of androgens.

[Morphofunctional characteristics of the thyroid in drug addicts].

Changes in the thyroid were studied of 34 patients who had died of heroin overdosing. Antibodies against hepatitic C virus were found in the serum of all the deceased. Alterations in thyroid tissue detected by morphological, histochemical and morphometric examinations correlated with serum content of hormones cT4, oT3 and TSH. The degree of structural changes in the thyroid correlated with duration of heroin use, its dose and its combination with other pharmacological drugs.

Hematological parameters in wistar rats in conditions of stress and treatment with heparin.

The effects on hematological measures of emotional tension arising during the formation of food-procuring behavior in a multiple-choice maze and in conditions of pain (injection of 0.85% physiological saline i.m., 0.3 ml, daily for five days) were studied in Wistar rats. Physiological saline and training of intact animals led to increases in RBC volume and mean corpuscular hemoglobin content, accompanied by significant increases in plasma corticosterone levels. At the same time, neither heparin (standard unfractionated heparin, Serva, doses of 64 and 640 IU/kg, daily for five days) nor training of heparinized rats produced any changes in these measures. The high levels of organization of the acquired behavior and the working capacity seen in heparinized rats were evidence for improvements in both the associative properties of conditioned reflex activity and the mechanisms supporting stable reproduction of the acquired habit. After administration of heparin, rats lost all types of manifestation reflecting psychoemotional tension as generally seen in animals in the active (administration of physiological saline) and passive (intact rats) controls.

[Morphological alterations of the rat myocardium in chronic morphine intoxication]. / Morfologicheskie izmeneniia miokarda krys pri khronicheskoi intoksikatsii morfinom.

Morphine was administered intraperitoneally to Wistar rats at increasing doses from 10 to 60 mg pay during 8 days. Cardiomyocyte damage in the form of myocytolysis and wave-like deformations of muscle fibers prevailed at the level of light microscopy. Microcirculatory alterations were characteristic: stasis, sludge-phenomenon, perivascular and interstitial oedema, diapedes hemorrhages, focal proliferation of vascular wall cells. Ultrastructural lesions were as follows: subsarcolemma edema, mitochondrial destruction, formation of giant forms of mitochondria, single contractures, increasing micropinocytosis, intestitial edema.

[Effects of stress agents and heparin administration on hematological parameters in Wistar rats]. / Gematologicheskie parametry u krys linii Vistar pri vozdeistvii stressornykh agentov i vvedenii geparina.

In chronic experiments in Wistar rats, the behavioral and hematological effects of the following stress factors were investigated: learning in complex maze and five multiple injections of the physiological saline (0.85%). Increasing of the erythrocyte volume and the contents of hemoglobin per erythrocyte in saline-induced groups and naive rats were observed after the learning. Moreover, the level of corticosterone in plasma of these rats was high. While in heparin-pretreated animals (unfractioned heparin, 64 and 640 IU/kg once per day, i.m., 5 days) these markers of stress are not revealed. The speed and efficiency of learning, behavior organization were significantly better in heparin pretreated rats than in naive and saline-induced groups. In heparin treated rats, a few types of unconditioned responses may be observed suggesting an anxiety or fear. We suppose that, after injections of heparin, complicated biochemical modifications of the rat organism that cause harmonization of inhibiting and excitatory processes of central nervous system, take place.

[Effect of folliculin and estradiol-dipropionate on the phagocytic properties of the blood neutrophils of the female white rat]. / Vliianie follikulina i éstradiol-dipropionate na fagotsitarnye svoistva neitrofilov krovi u samok belykh krys.

In the female rat blood, prior to and 5, and 50 days after bilateral ovarectomy with the substitutional administration of 50 units of folliculin and estradiol--dipropionate subcutaneously every 48 hrs for 50 days, a progressing decrease of the phagocytic activity was observed. Under the effect of estradiol--dipropionate the phagocytic activity reduced after ovarectomy was gradually returning to the initial level, whereas under the effect of folliculin the phagocytic activity increased proportionally to the time of incubation of blood with Staphylococcus aureus bacterial areas. The phagocytic activity of the blood neutrophiles seems to depend on estrogens.

[Diapedetic characteristics of the eosinophilic leukocytes in the uterus of white rats]. / Osobennosti diapedeza éozinofil'nykh leikotsitov v matke belykh krys.

Eosinophilic leucocytes, migrating in the uterine tissues during diestrus and proestrus, emigrate into the uterine cavity, being subjected to partial dystrophy. Dystrophy and assimilation of leucocytes, destroyed by the uterine tissues, especially by the endometrium, intensify and prevent emigration during proestrus and oestrus cycles. When the prior leucocyte destruction and assimilation are incomplete, phagocytosis of leucocytes and their fragments is arising, being realized by vacuolized cells of the uterine gland and epithelial lining, acquiring phagocytic properties under estrogen effect. This is the last barrier on their way to the uterine cavity, where the maximum pure genital tract flora is expedient during proestrus and oestrus. It is suggested that uterine leucocytes, representing carriers of biologically active compounds, including hormones, fulfil a transport function.