Intact Control Animals Research Articles

Quantitative morphological analysis of age structural changes in prostate of experimental animals with ethanol poisoning

Aim: To find out the age remodeling of the structural components of the prostate gland at alcohol poisoning using quantitative morphological analysis. Materials and Methods: The structure of the prostate gland of 4 white male rats groups were morphologically investigated. The 1 group included 30 control intact animals aged 8 months, the 2-nd group – 30 rats aged 24 months, the 3-rd group – 30 8-month-old animals with ethanol intoxication, and the 4-th group included 30 24-month-old rats with the specified simulated pathology. Ethanol intoxication was modeled by intragastric administration of 30% ethyl alcohol solution at a dose of 20 ml/kg once daily for 28 days. Rats were euthanized by bloodletting under general thiopental anesthesia 28 days after the beginning of the experiment. The area of glands, the height of glandular epithelial cells, the area of their nuclei and cytoplasm, the nuclear-cytoplasmic ratio in these cells and the stromal-parenchymal ratio in the organ were studied using light microscopy and were determined morphometrically. Morphometric parameters were processed statistically. Results: It was established that with age in the intact prostate of laboratory sexually mature white male rats, the area of glands, the height of glandular epitheliocytes, the area of their nuclei and cytoplasm, with the stability of nuclear-cytoplasmic ratios in the epithelial cells of the glands, significantly decreases, and the stromal-parenchymal ratio in the organ under study increases. Long-term ethanol poisoning leads to pronounced structural changes in the prostate, which is characterized by pronounced atrophy of the glandular epithelium, a decrease in the area of the glands, a decrease in the height of epithelial cells, a violation of nuclear-cytoplasmic relations in them, an increase in stromal-parenchymal ratio, and a prominent growth of the muscle-elastic stroma. The revealed structural changes of the studied components of the prostate dominated in 24-month-old experimental animals. Conclusions: Morphological analysis of the prostate gland established that morphometric and morphological changed significantly according to the age and were depend on the ethanol poisoning.

STUDY OF ENDOTHELIAL DYSFUNCTION INDICATORS IN RATS WITH EXPERIMENTAL PERITONITIS

Background. In the structure of purulent complications, peritonitis, destructive lesions of abdominal organs, and, as a rule, advanced forms of these diseases occupy one of the first places - 15-25% of urgent surgical diseases are complicated by peritonitis. It is generally known that the leading role in the triggering mechanism of the development of peritonitis belongs to the systemic inflammatory reaction, a component of which is phagocytosis, cellular and humoral immunity. However, the results of research on the state of functional activity of the endothelium in experimental peritonitis are limited.
 Aim: to study the activity of NO-synthase, FV and endothelin-1 in rats with experimental peritonitis.
 Materials and methods. Experimental studies were conducted on 24 non-linear laboratory rats, which were divided into 2 groups: 1 group – intact control (animals received distilled water), 2 group – animals of the control pathology group. According to the "Methodological recommendations for preclinical study of medicinal products", experimental peritonitis was studied on the model of V. A. Lazarenko. Indicators of vascular-platelet and coagulation hemostasis were determined according to generally accepted methods. Indicators characterizing endothelial dysfunction were studied according to generally accepted methods.
 Results. The most likely mechanism that is damaged in the endothelium during peritonitis is the activation of the synthesis of inducible NO-synthase by neutrophils/macrophages in response to infection. It is possible that the hyperproduction of nitric oxide, on the one hand, is aimed at destroying microflora and oxidizing toxins, and on the other hand, at suppressing the expression of tissue factor and cell adhesion molecules. platelet aggregation and cascade disorders in the hemostasis system. In animals with experimental peritonitis against the background of oxidative stress, there was an increase in the number of circulating desquamated endothelial cells in the blood, which is a highly specific marker of endothelial dysfunction. The level of the Willebrand factor also increased, which indicates the pathogenetic dependence of the factors that damage the endothelium of the vascular wall on the concentration of the Willebrand factor, which helps to reduce the permeability of blood vessels by adhesion of platelets to the endothelium. Confirmation of the development of endothelial dysfunction in peritonitis is an increase in the concentration of endothelin-1, which is a regulator of the process of vascular neoangiogenesis in response to endothelial damage.
 Conclusion. Hyperproduction of nitric oxide not only reflects the processes that occur in the focus of vascular endothelium damage, but also affects the severity of the inflammatory process and the consequences of the disease.

STUDY OF THE ROLE OF HEMOCOAGULATION DISBALANCE AS A KEY PATHOGENETIC PREDICTORS IN THE FORMATION OF SYSTEMIC DISORDERS IN EXPERIMENTAL PERITONITIS

Background. Peritonitis today continues to be one of the most difficult problems of both abdominal surgery and theoretical pathophysiology. One of the leading pathogenetic mechanisms of peritonitis is disorders in the hemostasis system and the development of dissemination vascular syndrome. Blockade of microcirculation caused by the development of dissemination vascular syndrome leads to tissue hypoxia and, as a result, to dysfunction of organs and body systems.
 Aim: is to study changes in vascular-platelet and coagulation hemostasis under conditions of experimental fecal peritonitis in rats.
 Materials and methods. Experimental studies were conducted on 24 non-linear laboratory rats, which were divided into 2 groups: 1 group – intact control (animals received distilled water), 2 group – animals of the control pathology group. According to the "Methodological recommendations for preclinical study of medicinal products", experimental peritonitis was studied on the model of V. A. Lazarenko. Indicators of vascular-platelet and coagulation hemostasis were determined according to generally accepted methods.
 Results. On the first day of experimental peritonitis, a sharp increase in the rate of blood clotting and changes in the quality properties of platelets were noted, affecting their aggregation ability and retraction of an already formed blood clot. On the 4th day of the experiment, changes in the vascular-platelet link were established, indicating the exhaustion of the platelet potential, due to a decrease in blood clotting time, platelet aggregation ability, and blood clot retraction against the background of a probable increase in the number of blood platelets, which probably by now had time "get" to the bloodstream from the depot. Changes in the coagulation hemostasis system against the background of experimental fecal peritonitis indicate the development of hypercoagulation already on the first day, which is replaced by a hypocoagulation state, which indicates the exhaustion of the enzymatic systems of blood coagulation potential (as early as on the 4th day), which lasted until the end of the experiment.
 Conclusion. In the development of disorders of the hemostasis system in fecal peritonitis, the key role belongs to both the internal and external mechanisms of activation of the blood coagulation system.

HISTOLOGICAL AND MORPHOLOGICAL CHANGES IN THE LYMPHOID STRUCTURES OF THE GASTRIC MUCOUS MEMBRANE IN WHITE RATS WITH THE ADMINISTRATION OF SODIUM GLUTAMATE.

The aim: To determine the histological and morphological changes of the lymphoid structures of the stomach in male rats under the influence of oral sodium glutamate at the rate of 15 mg/kg of body weight. Materials and methods: The scientific experiment was performed on 20 white non-linear male rats of reproductive age (4-5 months). The experimental animals were divided into two groups (10 rats in each group), which were orally received monosodium glutamate at a dose of 15 mg/kg body weight every day. We studied the effect of 2 and 4 weekly administration of monosodium glutamate at a dose of 15 mg/kg body weight, respectively, in the I and II groups of experimental animals (depending on the week of their decapitation). Rats of the control groups (n=10) were injected with a placebo for 2 and 4 weeks, namely 0.5 ml of dechlorinated tap water at room temperature. Intact control animals were also divided into two groups, 5 rats each, depending on the week of decapitation: respectively, III group - decapitation on the 2nd week of the experiment; IV group - decapitation on the 4th week of the experiment. After the experiments were completed, animals were decapitated under light ether anesthesia. According to the purpose of the study, pieces of rat stomach measuring 1.0 x 1.0 cm were taken from the front wall of the bottom of the stomach near the great curvature, cardiac and portal parts of the organ. Histological preparations were examined using a MICROmed SEO SСAN light microscope and a Vision CCD Camera. Morphometric studies were carried out according to the method of S. B. Stefanov, using grids No. 3/16. For electron microscopic examination, pieces of the stomach wall of rats were fixed in a 2.5% solution of glutaraldehyde in a 0.1 M phosphate buffer (pH 7.2-7.4) with subsequent fixation in a 2.0% solution of osmium tetroxide. After dehydration in alcohols and acetone, the material was embedded in eponaraldite. Sections were made on an LKB-8800-III ultramicrotome and studied using a JEM - 100-V microscope. To study the structural components of the lymphoid formations of the mucous membrane of different parts of the stomach of rats, semi-thin sections were made for the purpose of sharpening the blocks, which were stained with methylene blue. Results: The analysis of the obtained data of the conducted experiment indicates that the administration of monosodium glutamate in a dose of 15 mg/kg of body weight to rats already after 14 days leads to an increase in the density and size of the lymphoid structures of the GMM. The number of immunocompetent cells between the fundus of the gastric glands and the muscle plate increases in the diffuse lymphoid tissue of the gastrointestinal tract of rats in all its parts, both in the I and II groups of experimental animals. These changes are most pronounced in the cardiac and portal parts of the stomach. In both groups of experimental animals, the migration of interepithelial lymphocytes, macrophages, plasma cells, and tissue basophils to the surface epithelium increases. In both groups of experimental animals (and the II group of rats), lymphoid nodules and lymphoid pre-nodules of the gastric mucous membrane (GMM) are located between the bottom of the gastric glands and the muscular plate of the GMM. A gradual increase of medium lymphocytes in the GMM was established both in animals of I and II groups, while large lymphocytes increased in almost the same amount in experimental animals of both groups. Similar changes occur in the characteristics of the number of plasma cells, macrophages and tissue basophils in the lymphoid pre-nodules of GMM. Conclusions: Administering monosodium glutamate to rats at a dose of 15 mg/kg of body weight for 2 weeks leads to an increase in the density and size of lymphoid structures of the mucous membrane in all parts of the stomach with a predominant increase in the number of immunocompetent cells between the bottom of the gastric glands and the muscle plate. At the same time, more pronounced changes were found in the number of small lymphocytes, which tend to decrease by the 2nd week of the experiment, and vice versa - their density increases by the 4th week of monosodium glutamate administration.

COMPARISON OF EFFECTS OF FULLERENE C60 AND METHYL METHACRYLATE ON MASS INDEX OF INTERNAL ORGANS IN ANIMALS

In the clinic of orthopaedic dentistry, diseases of the mucous membrane caused by wearing removable dentures occupy a special place because the frequency of their occurrence and according to various authors varies from 15 to 70% of patients. Denture stomatitis is a complication resulting from wearing laminar dentures made of acrylates, which contain residual monomers having a chemical-toxic effect.
 The purpose of the study is to determine the mass coefficient of internal organs in order to assess the toxic effect of C60 fullerene on the body of rats and subsequent introduction of the material into the clinic of orthopedic dentistry.
 Objects and methods. The study was carried out on male rats divided into three groups of six animals each: I group included intact control rats; II group included animals received food with C60 fullerene; III group included animals received injectable methyl methacrylate. Conditionally therapeutic dose of fullerene C60 and methyl methacrylate was 1 ml/kg each. The studied samples were administered to rats through a metal probe every day once a day for 21 days.
 The state of peripheral blood, the functional state of the liver and the mass coefficient of the internal organs of the studied animals enable to evaluate and compare the toxic effect of fullerene C60 and methyl methacrylate.
 Results. The study has shown the introduction of C60 fullerene did not cause visible signs of intoxication and lethal consequences in the experimental animals. No animals of the main group (fullerene C60) and the comparison group (methyl methacrylate) died after the 21-day study. There were also no significant violations of the general condition and behaviour of the animals. The physiological state of experimental rats did not differ from control rats.
 In the group of intact control males, a significant increase in the body weight of rats compared to the initial values ​​was noted by 8% in the 2nd week and by 20% in the 3rd week of observation. In groups of animals that were injected with fullerene C60 and methyl methacrylate, the body weight dynamics as a whole was similar to the body weight dynamics of intact control animals. The obtained indicators point out the absence of a harmful effect of the studied drug on the physiological processes of rats.
 Administration of the nanopreparation did not have a harmful effect on internal organs (liver, heart, lungs, spleen, adrenal glands), with the exception of the kidneys and thymus, where certain changes were observed. The relative mass of the kidneys decreased statistically significantly when fullerene C60 was used, and the mass of the thymus, on the contrary, increased. Despite the statistical differences between the groups, the values ​​of the kidney mass coefficient for the use of the nanopreparation are within the physiological range and cannot be considered critical. An increase in the mass of the thymus against the background of fullerene use probably occurs as a result of the activation of the immune system upon the introduction of the investigated agent. However, hematological studies did not reveal pathological deviations in the leukocyte blood formula and significant activation of immunological processes, so it can be assumed that the increase in thymus mass coefficient is not a consequence of the toxic effect of C60 fullerene.
 Conclusion. The use of Fullerene C60 for 21 days, based on the results obtained, does not have a toxic effect on the organs and systems of male rats, does not cause any intoxication reactions and does not lead to fatal consequences.

The Protective Effects of S-Methyl Methionine Sulfonium Chloride on Brain Tissue Damage in D-Galactosamine-Induced Hepatotoxicity

Objective: The objective of the current work is to examine the protective effects of S-methyl methionine sulfonium chloride (MMSC) on brain in galactosamine (GalN)-induced hepatotoxicity in rats. Materials and Methods: A total of twenty two female Sprague-Dawley rats were randomly assigned into four groups as follows: Group I (n=5), intact control animals; Group II (n=6), animals that received 50 mg/kg/day of MMSC by gavage technique for 3 consecutive days; Group III (n=5), animals injected with a single dose of 500 mg/kg of GalN intraperitoneally (i.p.); and Group IV (n=6) are animals injected with the same dose of GalN (i.p.) 1 hour after MMSC treatment. At the end of the experiments (after 6 hours of the last GalN treatment), all animals were sacrificed under anaesthesia, and brain tissues were dissected out. Results: A statistically remarkable increase in lipid peroxidation, hydroxyproline, and nitric oxide levels, was detected while a notable decline in the activities of sodium/potassium ATPase was observed in GalN group in comparison with control rats. In contrast, all alterations observed were reversed when MMSC was given to GalN groups. Conclusion: Consequently, it may be considered that MMSC has a protective role on brain in GalN-induced hepatotoxicity in rats.

Monocrotaline‐induced Pulmonary Hypertension Impairs Diaphragm Vasomotor Function

INTRODUCTIONPulmonary hypertension (PH) is a chronic and progressive disease characterized by high arterial blood pressure and compensatory vascular remodeling in the pulmonary circulation. Mounting evidence suggests that the effects of PH extend beyond the pulmonary circulation. In humans and animals, PH induces diaphragm contractile dysfunction and diaphragm muscle atrophy. While the mechanistic bases for the PH‐induced diaphragm dysfunction is relatively unknown, alterations in the vasomotor control of the diaphragm resistance vasculature (i.e. arterioles) may play a key role. Therefore, we tested the hypothesis that PH increases the vasoconstrictor responsiveness and impairs the vasorelaxation of diaphragm arterioles.METHODSAdult female Sprague‐Dawley rats (n=8) were divided into healthy control (HC; n=2), ovariectomized (OVX; n=3), and PH + OVX (n=3). PH + OVX animals were given a single intraperitoneal injection of monocrotaline (MCT; 50mg/kg) to induce PH. Disease progression was then monitored via echocardiography. Once animals developed PH (~3 weeks post injection), the diaphragm was removed and first‐order arterioles (1A) from the medial costal diaphragm were isolated, cannulated, and pressurized at 90 cmH2O. After a 60 min equilibration period and spontaneous tone development, vasoconstrictor responses to potassium chloride (KCl) (10‐100 mM) and endothelium‐dependent and ‐independent vasorelaxation responses to cumulative doses of acetylcholine (ACh) and sodium nitroprusside (SNP) (10‐9 to 10‐4 M) were determined.RESULTSSpontaneous tone (%) development was not different across all three groups (HC: 36 ± 9%; OVX: 33 ± 9%; PH + OVX: 39 ± 2%; P>0.05). Compared to control vessels, maximal percent vasoconstriction with KCl was decreased with OVX (67 ± 2 versus 50 ± 1%; P<0.05). Maximal vasoconstriction (%) was reduced with PH + OVX (36 ± 4%) compared to vessels from both healthy control and OVX groups (P<0.05). Ovariectomy had no effect on endothelium‐dependent vasodilation responses to ACh versus intact control animals (P>0.05). The maximal ACh‐mediated vasodilation with PH + OVX (29 ± 9%) was decreased versus both healthy (45 ± 6%; P<0.05) and OVX animals (41 ± 4%; P<0.05). Across all three groups, there were no differences in percent relaxation with SNP (P>0.05).CONCLUSIONThese findings suggest that estrogen plays a role in the maintenance of K+ channel activity and/or the contribution of K+ channels to diaphragm vasomotor tone, and that PH further disrupts K+ channel function in diaphragm arterioles. In addition, the absence of estrogen in OVX animals had no effect on endothelial function whereas PH + OVX impaired endothelium‐dependent vasorelaxation of diaphragm resistance vessels. These findings suggest that PH, at least in the rodent model utilized herein, elicits endothelial dysfunction in the diaphragm vasculature and provide a potential mechanism for the impaired diaphragm contractile dysfunction in PH patients.

The study of amino acids level in the vitreous body of experimental animals in regamatogenous retinal detachment

INTRODUCTION: The paper presents the results of the amino acid levels study in the vitreous body of rats with rhegmatogenous retinal detachment (RRD). MATERIAL AND METHODS: Experimental studies were performed on 42 brown Norwegian male rats (Male Brown Norway), which were divided into 7 groups (6 animals in each group): 1 group — conditionally intact control (without retinal detachment) — animals that underwent paracentesis of the anterior chamber with the removal of its moisture and retinal puncture without the introduction of any substance under the retina; 2 group — rats, which reproduced RRD by the method (control pathology) based on the study of apoptosis induction by apoptosis-inducing factor. RESULTS: In the group of conditionally intact control animals the concentration of amino acids in vitreous body were: alanine — 3.3 ± 0.27 ng/mL; arginine — 0.8 ± 0.1 ng/mL; aspartate — 2.3 ± 0.5 ng/mL; valine — 1.8 ± 0.8 ng/mL; histidine — 1.3 ± 0.3 ng/mL; glycine — 4.5 ± 0.8 ng/mL; glutamic acid — 1.56 ± 0.35 ng/mL; tyrosine — 0.084 ± 0.048 ng/mL; phenylalanine — 0.024 ± 0.017 ng/mL; methionine — 0.11 ± 0.1 ng/mL. In animals with model pathology of RRD was observed a significant increase in certain amino acids: the level of alanine increased 1.4 times, aspartate — 11.5 times (p < 0.05), glycine — 2.3 times (p < 0.05), glutamic acid — 7.9 times (p < 0.05) compared with rats of the conditionally intact group. Levels of other amino acids increased insignificantly. CONCLUSIONS: The most pronounced therapeutic efficacy was found in rats treated with combination therapy with dexamethasone, resveratrol, erythropoietin, and edaravon for 7 days. The pronounced effect of combination therapy on the level of amino acids (aspartate, glutamate, alanine, and glycine) in the treatment of RRD is due to the pharmacological activity of the components of this therapeutic regimen and characterized by synergistic effects of each component in the key links of disease pathogenesis.

Evaluating Some Parameters Of Wistar Rat Brain In Traumatic Brain Injury Model With Administration Of Proline-Containing Peptides

Objective — To examine the morphometric parameters of neurons and the oxidative status of the Wistar rat brain tissues after administering proline-containing peptides, also known as glyprolines (Arg-Gly-Arg-Pro-Gly-Pro [RGRPGP] and Thr-Lys-Pro-Arg-Pro-Gly-Pro [WKPRPGP; Selank]), on a traumatic brain injury (TBI) model. Study subjects — Our study involved 26 mature male Wistar rats (2.5-3 mos. old, 220-300 g). The morphometric parameters of neurons and the oxidative status of animal brain tissues were studied. Material and Methods — Four experimental groups were formed. Group 1 included intact control animals. Animals in three other groups were subjected to TBI via free fall of a 50 g weight from a height of 50 cm on the second day of the experiment and received the following injections: 0.9% sodium chloride solution in Group 2; WKPRPGP peptide solution in Group 3; RGRPGP peptide solution in Group 4. Substances were administered intraperitoneally on a daily basis at a dose of 0.1 mg/kg from day 1 through day 5 of the experiment. Morphometric parameters of rat brain neurons were studied on paraffin sections stained with hematoxylin and eosin. The intensity of free radical processes in the brain tissue was investigated by chemiluminescence. Results — An analysis of morphometric parameters revealed significant increases in the neuronal cytoplasm area, nucleolar area, number of nucleoli, and nuclear-nucleolar index with the introduction of the RGRPGP peptide after TBI, compared with the WKPRPGP peptide under the same conditions. After TBI and peptide administration, we observed an oxidative stress in the neocortex of Wistar rats, and it was more pronounced in the group of animals treated with RGRPGP. Conclusion — After RGRPGP peptide administration, we observed an increase in the morphometric parameters of neurons in the closed TBI model: a larger area and a greater number of nucleoli. Chemiluminescence data implied that WKPRRPGP peptide better protected brain tissue in rats from the effects of oxidative stress caused by TBI.

Intraperitoneal administration of lipopolysaccharide (LPS) causes retinal ganglion cell loss in mice

AbstractPurposeTo analyze the effect of a systemic inflammatory insult induced by LPS administration on retinal ganglion cells and on microglial activation in female and male mice.MethodsFemale and male C57BL/6J mice were treated with a single intraperitoneal injection of LPS (5mg/kg). As control intact animals were used. At 3, 7 and 15 days after LPS administration, retinas were dissected as flat mounts and Iba1 (microglia) and Brn3a (RGCs) immunodetected.ResultsMicroglial activation was observed at 3 days and was qualitatively similar in both sexes. RGC loss was observed at 3 days (p < 0.0001 compared to intact retinas) and was significantly lower in females (13% loss of the original RGC population) than in males (20%). In both sexes, RGC loss progressed until day 7 reaching 28% in females and 34% in males. From 7 to 15 days there was not further loss of RGCs.ConclusionsOne single injection of LPS causes microglial activation and neuronal death that is higher in males than females.

In Vivo Antidepressant Efficacy of 3-Substituted Thietane-1,1-dioxide Derivative - A Preliminary Study for Novel Anti-Depression Therapy in Neurological Disorders.

Psychosocial stress-induced depressive behavior is linked to the etiology of several neurological diseases viz., PTSD, and neurodegenerative disease like Alzheimer's Disease (AD). The repeated bouts of social stress defeat can be induced using Resident-Intruder- Paradigm (RIP) and Chronic Mild Social Stress (CMSS) animal models to assess the stress-induced depressive behavioral patterns. The aim of this study to examine the anti-depressive efficacy of 3-methoxythietane- 1,1-dioxide (N-14) in RIP models of behavioral alterations. In this study, we have used Sprague-Dawley rats in Resident-Intruder-Paradigm (RIP), where intruders interacted with residents Day 0 to Day +5 for 10 minutes to invoke CMSS in intruders and became defeated/submissive rats due to the depressive-like behavioral alterations in social activity, explorations, grooming, defense, aggressive behavior, social interaction, freeze, rearing etc., with residents. Control intact animals are included in group I, group II received N-14 alone; group III received CMSS, and group IV received cotreatment of N14 with CMSS. N-14 (2 mg/kg) was administered intraperitoneally from Day 0 to Day +5 to intact animals and intruder animals under conditions of CMSS. Several behavioral tests viz., forced swim test, open field test, and elevated-plus maze test were used to examine the above behavioral dynamic parameters. The dynamic interaction between Residents and Intruders during the study showed substantial alterations in exploratory activity, aggressiveness, defensive behavior, body weight, and thymus mass in stressed animals. N-14 cotreatment has mitigated sociability, exploratory activity, aggressiveness increased social adaptability and defensive behavior. An extensive rise in active forms of defense and submission latency indicates that N-14 has induced antidepressant activity with a psycho-sedative component of action. Serendipitously, we observed the ameliorative capability of N-14 cotreatment to mitigate depressive-behavioral symptoms in intruders.

The Effect of Betulin Diphosphate in Wound Dressings of Bacterial Cellulose-ZnO NPs on Platelet Aggregation and the Activity of Oxidoreductases Regulated by NAD(P)+/NAD(P)H-Balance in Burns on Rats.

The inhibition of platelet aggregation, and the activity of oxidoreductases and microhemocirculation in a burn wound on the treatment of burns with wound dressings based on bacterial nanocellulose (BC)-zinc oxide nanoparticles (ZnO NPs)-betulin diphosphate (BDP) were studied. The control of the treatment by BC-ZnO NPs-BDP on burned rats by the noninvasive DLF method showed an increase in perfusion and the respiratory component in wavelet spectra, characterizing an improvement in oxygen saturation in the wound. The study on the volunteers’ blood found the inhibition of ADP-induced platelet aggregation by 30–90%. Disaggregation depends on the dose under the action of the ionized form of BDP and ZnO NPs-BDP in a phosphate buffer; it was reversible and had two waves. It was shown on rats that the specific activity of LDHreverse and LDHdirect (control-intact animals) on day 21 of treatment increased by 11–38% and 23%, respectively. The LDHreverse/LDHdirect ratio increased at BC-ZnO NPs-BDP treatment, which characterizes efficient NAD+ regeneration. AlDH activity increased significantly in the first 10 days by 70–170%, reflecting the effectiveness of the enzyme and NAD+ in utilizing toxic aldehydes at this stage of burn disease. The activities of GR and G6PDH using NADP(H) were increased with BC-ZnO NPs-BDP treatment.

Understanding molt control switches: Transcriptomic and expression analysis of the genes involved in ecdysteroidogenesis and cholesterol uptake pathways in the Y-organ of the blue crab, Callinectes sapidus.

The crustacean molting process is regulated by an interplay of hormones produced by the eyestalk ganglia and Y-organs (YO). Molt-inhibiting hormone and crustacean hyperglycemic hormone released by the sinus gland of the eyestalk ganglia (EG) inhibit the synthesis and secretion of ecdysteroid by the YO, hence regulating hemolymph levels during the molt cycle. The purpose of this study is to investigate the ecdysteroidogenesis pathway, specifically genes linked to changes in ecdysteroid levels occurring at early premolt (ePM). To this end, a reference transcriptome based on YO, EG, and hepatopancreas was de novo assembled. Two genes (cholesterol 7-desaturase Neverland and cytochrome p450 307a1-like Spook) involved in ecdysteroidogenesis were identified from the YO transcriptome using sequence comparisons and transcript abundance. Two other candidates, Hormone receptor 4 and probable cytochrome p450 49a1 potentially involved in ecdysteroidogenesis were also identified. Since cholesterol is the ecdysteroid precursor, a putative cholesterol carrier (Apolipoprotein D-like) was also examined to understand if cholesterol uptake coincided with the increase in the ecdysteroid levels at the ePM stage. The expression level changes of the five candidate genes in the YO were compared between intermolt (IM) and induced ePM (iePM) stages using transcriptomic analysis. Expression analysis using qPCR were carried out at IM, iePM, and normal ePM. The increase in Spook and Neverland expression in the YO at the ePM was accompanied by a concomitant rise in ecdysteroid levels. The data obtained from iePM stage were congruent with those obtained from the normal ePM stage of intact control animals. The present findings support the role of Halloween genes in the ecdysteroidogenesis and molt cycle in the blue crab, Callinectes sapidus.

Effect of liraglutide dose on efficacy of correcting microcirculatory disorders in absolute insulin deficiency of albino rats

The objective was to study the dose-dependent effects of glucagon-like peptide-1 receptor agonist, liraglutide, on microcirculation in white rats under the alloxan-induced insulin deficiency. Materials and Methods. Our study was carried out on 70 white rats, divided into four groups: 20 intact control animals; 20 animals of the comparison group with alloxan-induced diabetes; 10 animals with insulin deficiency, injected with liraglutide at a dose of 0.2 mg/kg per day (treatment group No. 1); and 20 animals with insulin deficiency treated with liraglutide at a dose of 0.4 mg/kg per day (treatment group No. 2). Diabetic status, skin perfusion of the posterior limb and mechanisms of blood flow modulation were assessed in all groups of animals. Results. It was established that administering liraglutide at a dose of 0.4 mg/kg per day allowed achieving complete normalization of carbohydrate metabolism in rats with alloxan-induced insulin deficiency, thereby more effectively improving the state of microcirculation, compared with a dose of 0.2 mg/kg per day. Conclusion. The efficacy of correcting the microcirculatory disorders with liraglutide correlates with correcting the carbohydrate metabolism in rats with insulin deficiency and depends on liraglutide dose. The dose-dependent effect is achieved by the endothelial mechanism of flow modulation in microcirculation. However, the restoration of the vascular tone neurogenic component is independent of liraglutide dose.

Vitamin U prevents valproic acid-induced liver injury through supporting enzymatic antioxidant system and increasing hepatocyte proliferation triggered by inflammation and apoptosis

The aim of this study was to investigate the cellular mechanisms that cause valproic acid (VPA)-induced liver damage and the therapeutic effect of Vitamin U (Vit U) on these mechanisms. Female Sprague Dawley rats were randomly divided into four groups: intact control animals, animals that received Vit U (50 mg/kg/day), animals given VPA (500 mg/kg/day), and animals given both VPA and Vit U. The rats in the Vit U + VPA group were administered Vit U by gavage an hour before VPA administration every day for 15 days. Liver tissues were evaluated through histopathological, biochemical, immunohistochemical, and Western blotting techniques. Administration of Vit U with VPA resulted in (i) prevention of histopathological changes caused by VPA; (ii) blockage of the decrease in catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), and superoxide dismutase (SOD) activities; prevention of the elevation in gamma-glutamyl transferase (GGT) activity and advanced oxidation protein products (AOPP) level; (iii) increased in the levels of interleukin-1 beta (IL-1β), active caspase-3, and cytoplasmic cytochrome c; (iv) increase in cleaved poly (ADP-ribose) polymerase (PARP) level and decrease in LC3B (II/I) ratio; (v) increase in the number of proliferating cells nuclear antigen (PCNA) positive hepatocytes. These findings show that Vit U prevents liver damage caused by VPA through increasing the antioxidant enzyme capacity and hepatocyte proliferation by triggering inflammation and apoptosis. These findings suggest that Vit U provides its protective effects against VPA-induced liver damage by stimulating homeostasis and regeneration.

The Effects of NPY1 Receptor Antagonism on Intervertebral Disc and Bone Changes in Ovariectomized Rats.

Study Design:Basic science.Objective:To compare the effects of a neuropeptide Y1 receptor antagonist (NPY-1RA) to estrogen on maintaining vertebral bone microarchitecture and disc height in a rat model of menopause.Methods:This study was an institutional animal care approved randomized control study with 104 ovariectomized rats and 32 intact control animals. Comparison of disc height, trabecular bone, body weights, circulating levels of NPY and estrogen, and distribution of Y1 receptors in the intervertebral disc in an established rodent osteoporotic model were made at baseline and after 2, 4, and 8 weeks after receiving either an implant containing estrogen or an antagonist to the neuropeptide Y1 receptor. Data was compared statistically using One-way analysis of variance.Results:Circulating levels of estrogen increased and NPY decreased following estrogen replacement, with values comparable to ovary-intact animals. NPY-1RA-treated animals had low estrogen and high NPY circulating levels and were similar to ovariectomized control rats. Both NPY-1RA and estrogen administration were able reduce, menopause associated weight gain. NPY-1RA appeared to restore bone formation and maintain disc height, while estrogen replacement prevented further bone loss.Conclusion:NPY-1RA in osteoporotic rats activates osteoblast production of bone and decreased marrow and body fat more effectively than estrogen replacement when delivered in similar concentrations. Annulus cells had NPY receptors, which may play a role in disc nutrition, extracellular matrix production, and pain signaling cascades.

ANTIOXIDANT POTENTIAL OF ADEMETIONINE IN GENTAMICINE NEPHROPATHY PROGRESSION

Acute kidney injury has been globally considered as a serious medical issue. Its incidence rate is increasing over the years despite the introduction of new therapies, the cause of which is often a side effect to drugs, or due to the impact of toxins, trauma. Nephrotoxicity is one of the most important side effects of aminoglycoside antibiotics, and on gentamicin, in particular. Despite rigorous patient monitoring, this type of nephrotoxicity appears in 10–30% of therapeutic courses. Therefore, searching for new therapies, which would be able to prevent or decrease toxic kidney injury during the treatment with aminoglycosides is quite relevant. Ademetionine is an amino acid derivative, which possesses various effects such as antioxidant, membrane stabilizing, anti-inflammatory and regenerative.
 Aim of the research is to study an antioxidative activity of ademetionine in conditions of gentamicin nephropathy progression in rats. Materials and methods. Research was conducted on non-linear mature white rats weighting 130-180 g, randomly divided into 3 groups (n = 7): I group included intact control animals, II group included rodents with gentamicin nephropathy (injection of 4% Gentamicin sulphate solution in a dosage of 80 mg/kg for 6 days), III group involved the rats receiving ademetionine («Abbott SpA», Italy) in a dosage of 20 mg/kg. Peroxidation processes in blood and kidneys were evaluated by the content of active products that react with thiobarbituric acid and oxidative modification of proteins levels, antioxidant defence was assessed by catalase and glutathione peroxidase activity, SH-groups and ceruloplasmin content.
 Results. In the course of experimental studies on the model of gentamicin nephropathy, the expressive antioxidant activity of ademetionine was proven: it was demonstrated by reducing the intensity of lipid peroxidation (a decrease in the content of active products that react with thiobarbituric acid) and proteins (a decrease in the content of oxidative modified proteins) in the blood and kidney homogenate, next with increased activity of the enzymatic link of antioxidant protection (glutathione peroxidase and catalase) and non-enzymatic (ceruloplasmin and compounds with SH-groups). The antioxidant potential of ademetionine has been confirmed by an increase in the antioxidant-prooxidant index in kidney tissue and a significant decrease in the index of oxidative stress in the blood of treated animals.
 Conclusion. Thus, the results of experimental studies indicate the antioxidative effect of ademetionine in modelled gentamicin nephropathy, and point out the necessity of further studying its potential nephroprotectective properties.

АКТИВНІСТЬ СУКЦИНАТДЕГІДРОГЕНАЗИ МІТОХОНДРІЙ ПЕЧІНКИ ТВАРИН-РЕЦИПІЄНТІВ ЗА ВПЛИВУ АЛОГЕННИХ МЕЗЕНХІМАЛЬНИХ СТОВБУРОВИХ КЛІТИН

The studies were conducted on 2-3-months-old males of C57BL/6 mice weighing 20-24 g. О btaining of allogenic bone marrow and adipose derived MSCs were carried out in a sterile laminar box. Cells were cultivated in a CO 2 -incubator at 37 о C and 5 % CO2 in DMEM with 10-15 % of fetal bovine serum, 1% of antibiotic-antimycotic solution (Sigma-Aldrich, USA). The following groups of animals were formed: 1 group – intact (control) animals; 2 group – animals, to whom 0.5 ml of 0.89% NaCl solution (placebo) were injected into the caudal vein; 3 group – animals, to whom 10 4 of allogenic AD MSCs in 0.5 ml of phosphate buffer solution were injected into the caudal vein, 4 group – animals, to whom 10 4 of allogenic BM MSCs in 0.5 ml of phosphate buffer solution were injected into the caudal vein. On 12th day after transplantation of MSCs activity of succinate dehydrogenase in animals were determined. The determination of the activity of the enzyme was carried out in accordance with the method, the principle of which is the restoration of potassium ferricyanide (K 3 [Fe(CN) 6 ]) to potassium ferrocyanide (K 4 [Fe(CN) 6 ]) by succinate with involvement of succinate dehydrogenase . Activity was determined by the volume of recovered ferricyanide. Statistical processing of the results was performed with using of Origin 6.1 software and Student's t-criterion. All data are presented in the form of arithmetic average and standard deviations. It was found that at the 12 day of the study, the activity of the succinate dehydrogenase in the third experimental group was 57.7 ± 1.6 mmol / l K 3 [Fe (CN) 6 ] / mg * min (p <0.001), which is significantly higher than in the animals of the first and second group (45.9 ± 0.7 and 43.3 ± 1.2 mmol / l K 3 [Fe (CN) 6 ] / mg * min, respectively). The activity of the enzyme in animal of the fourth experimental groups was also reliable higher compared to the first two groups and was 53.3 ± 1.4 mmol/l K 3 [Fe(CN) 6 ] / mg * min (p < 0.01). It should be noted that mitochondrial succinate dehydrogenase activity in the liver of recipient animals after transplantation of MSCs from adipose tissue is significantly higher than after transplantation of MSCs, obtained from bone marrow (p < 0.05). Thus, reliable raising of mitochondrial succinate dehydrogenase activity in the liver of recipient animals after transplantation of allogenic mesenchymal stem cells from adipose tissue and bone marrow has been determined. Keywords : mesenchymal stem cells, mitochondria, succinate dehydrogenase, mice, bone marrow, fatty tissue.

Hippocampal Damage Causes Retrograde Amnesia and Slower Acquisition of a Cue-Place Discrimination in a Concurrent Cue-Place Water Task in Rats

Explanations of memory-guided navigation in rodents typically suggest that cue- and place-based navigations are independent aspects of behavior and neurobiology. The results of many experiments show that hippocampal damage causes both anterograde and retrograde amnesia (AA; RA) for place memory, but only RA for cue memory. In the present experiments, we used a concurrent cue-place water task (CWT) to study the effects of hippocampal damage before or after training on cue- and place-guided navigation, and how cue and place memory interact in damaged and control rats. We found that damaging the hippocampus before training caused a delay in the expression of cue-place navigation strategies relative to intact control animals; surprisingly, place navigation strategies emerged following pre-training hippocampal damage. With additional training, both control and damaged rats used local cues to navigate in the CWT. Damaged animals also show minor impairments in latency to navigate to the correct cue following a cue contingency reversal. By contrast to these anterograde effects, damage made after training causes RA for cue choice accuracy and latency to navigate to the correct cue. In addition, the extent of hippocampal damage predicted impairments in choice accuracy when lesions were made after training. These data extend previous work on the role of the hippocampus in cue and place memory-guided navigation, and show that the hippocampus plays an important role in both aspects of memory and navigation when present during the learning experience.

Increased Hematocrit Due to Electrical-Waveform Exposures in Splenectomized Sus scrofa.

In laboratory studies of the pig Sus scrofa, hematocrit has consistently increased after conducted-electrical-weapon (CEW) exposures, possibly due to contraction of the spleen. Splenectomized animals and intact sham control animals were exposed, each for 30 sec, to a benchtop-produced electrical waveform of net charge levels similar to those of some CEWs. Changes in the blood were compared statistically. Hematocrit increased significantly in both splenectomized and sham animals. There were no significant main-effect differences between values of hematocrit from the two groups. There were, however, significant interactive effects of time and splenectomy for hematocrit, red blood cell count, and hemoglobin. After peak values were reached for these variables, values returned toward baseline levels more slowly in splenectomized animals. This may have been due to the lack of a spleen to sequester red blood cells (thereby resulting in more cells remaining in the general circulation), unlike sham animals with intact spleens.