Peculiarities of morphological changes of endotheliocytes and remodeling of the arteries under the experimental hypercholesterolemia.

OBJECTIVE: Introduction: Under the conditions of experimental hypercholesterolemia, endothelial dysfunction develops with the morphological marker which is an increase in the number of blood-circulating desquamated endothelial cells (DEC), but this situation needs to be clarified in the development of this pathology in the age aspect. The aim: To find out the features of remodeling of endothelial cells and arteries of the hind limbs in the rats of pre-repopductive and reproductive age with experimental hypercholesterolemia. PATIENTS AND METHODS: Materials and methods: The experimental group consisted of 16 animals with biochemically confirmed hypercholesterolemia, which were divided into 2 groups: group 1 - 8 animals, aged 2-3 months, weighing 150-170 grams and group 2 - 8 rats aged from 11 to 11 months weighing 230-250 gram. The control group consisted of rats of the same age of 8 animals in each. RESULTS: Results: Hypercholesterolemia causes damage to the vascular endothelium of the arteries, which is characterized by an increase in the number of desquamated endothelial cells in the peripheral blood. The most circulating blood in desquamated endothelial cells was detected in 45 days of study in animals of reproductive age, where the number of desquamated endothelial cells increased by 2.56 times, and in animals of pre-reproductive age - 2.35 times. Morphological changes were characterized by thickening of the intima of the arteries of the femur, knee and tibia due to swelling of the endothelial cells, their desquamation and proliferative changes in places of preserved vascular endothelium. In response to the deposition of lipids and PAS-positive substrates, cellular reactions appeared as weak lymphocytic infiltration. In addition to hyperlastosis, fragmentation of elastic fibers was revealed. Correlation of intima contributed to the narrowing of vascular lumen. Lipids, xanthoma cells and sour mucopolysaccharides were accumulated in the inner membrane of the arteries. In addition to lymphocytic infiltrates, the amount of collagen fibers in adventitia increased. CONCLUSION: Conclusions: Under conditions of hypercholesterolemia the number of desquamated endothelial cells in the blood increases, and arterial remodeling is characterized by manifestations of hypertrophic-neoplastic remodeling in rats of pre-reproductive age, and in reproductive animals there were sclerotic and inflammatory changes.

[Analysis of final judgements in cases of medical negligence occurred in Ukraine].

OBJECTIVE: Introduction: The peculiarities of the disadvantages of providing medical care in Ukraine are not well-known abroad. The aim: To study the peculiarities of court decisions in cases of unfavorable consequences of medical activity. PATIENTS AND METHODS: Materials and methods: The article analyzes the official data of the General Prosecutor's Office of Ukraine and the website of court decisions regarding criminal cases against medical practitioners. RESULTS: Review: Approximately 600 cases of alleged medical malpractice cases are registered annually in Ukraine. Only less than one percent of them are brought to the court. The guilt of medical practitioners was proven in majority (80,8%) of court decisions. Acquittals of defendants were pronounced in 5,9% of court verdicts. Obstetrics and gynecology, surgery, internal medicine and anesthesiology are in the top of high-risk medical specialties. CONCLUSION: Conclusions: Majority of medical malpractice litigations are sued in Ukraine baselessly. In cases of medical negligence majority of defendants are acquitted as usual.

Cerium dioxide nanoparticles possess anti-inflammatory properties in the conditions of the obesity-associated NAFLD in rats.

BACKGROUND: Obesity is a risk factor for non-alcoholic fatty liver disease (NAFLD). The disease is associated with impairment of pro/antioxidant equilibrium and the inflammation in liver tissue. The aim of the work was to investigate the anti-inflammatory properties of the nanocrystalline cerium dioxide (nCeO2) on the rat model of NAFLD associated with monosodium glutamate (MSG)-induced obesity. METHODS: The study was carried out on three groups of rats: control, MSG- and MSG+nCeO2. They were injected with saline (control) or MSG. A month after born MSG-rats had been treated with water in a volume of 2.9ml/kg, MSG+CeO2 groups - with CeO2 intragastrically (i.g.). The anthropometric and carbohydrate metabolism parameters, content of proinflammatory cytokines (interleukin (IL)-1ß, IL-12Bp40, interferon-γ (INF-γ)) and anti-inflammatory cytokines (IL-4, IL-10, tumor growth factor-ß (TGF-ß)) were measured by ELISA. RESULTS: We have demonstrated the anti-obesity effect of nanocrystalline cerium dioxide and for the first time its anti-inflammatory properties. Nanoparticles reduced the content of pro-inflammatory cytokines (IL-1ß, IL-12Bp40) in rat serum and restored the level of anti-inflammatory cytokines (IL-4, IL-10, TGF-ß) to the control values. CONCLUSION: The precise mechanisms of this phenomenon remain to be unclear but we suppose they are at least partially associated with the strong anti-oxidant action of studied substance. Nanocrystalline cerium dioxide attenuates the inflammatory processes in rat blood that can prevent obesity complications and liver injury.

Effects of polyphenol compounds melanin on NAFLD/NASH prevention.

BACKGROUND: One of the pathogenic mechanisms of the progression non-alcoholic liver disease (NAFLD) to nonalcoholic steatohepatitis (NASH) is the accumulation of reactive oxygen species (ROS). So, antioxidant therapy is necessary for successful treatment of the liver injury. We have paid attention to melanin produced by yeast Nadsoniella nigra strain X-1 as novel antioxidant and anti-inflammatory agents with low toxicity. In current study we aimed to investigate the preventive effect of melanin on the monosodium glutamate (MSG) induced NAFLD model in rats. METHODS: The study was carried out on 45 Wistar rats that were divided into 3 groups: intact, MSG- and MSG+melanin groups (n=15 in each group). Newborn rats of MSG- and MSG+melanin groups were administered with MSG (4mg/g, 8µl/g, subcutaneously) at 2nd-10th days of life. Since the age of 1 month, rats of MSG-group were treated with water (0.25ml/100g), rats of MSG+melanin groups-with melanin (1mg/kg) dissolved in water (0.25ml/100g). INTRODUCTION: had been performed intermittently (two-week courses alternated with two-week breaks) for 3 months. In 4-month rats anthropometrical parameters and visceral adipose tissue (VAT) mass were estimated. To assess morphological changes in liver we used NAS (NAFLD activity score). The content of pro-inflammatory cytokines (interleukin (IL)-1ß, IL-12Bp40, interferon (INF)-γ) and anti-inflammatory cytokines (IL-4, IL-10, tumor growth factor (TGF)-ß) were measured by ELISA. RESULTS: We found significantly lower total score (1.0±0.19 vs 3.33±0.36, p<0.001), degree of steatosis (0.73±0.18 vs 1.80±0.17, p<0.001) and manifestation of lobular inflammation (0.27±0.11 vs 1.20±0.17, p<0.001) due to NAFLD activity score in MSG+melanin group compared to MSG-obesity. NASH we confirmed only in 33.3% of rats with MSG-obesity that was significantly higher than after melanin (6.7%) administration (p=0.033). Melanin administration reduce amount of visceral fat on 44.5% (p<0.001) as compared to MSG-obesity group. Melanin reduced the content of IL-1ß in rat serum and restored the level of anti-inflammatory cytokines (IL-10, TGF-ß) to the control values. CONCLUSION: Thus, the administration of melanin can prevent development of NAFLD/NASH in rats with MSG-induced obesity and can be considered as possible novel therapeutic agents but further studies to confirm its action needed.

Probiotics Supplemented with Omega-3 Fatty Acids are More Effective for Hepatic Steatosis Reduction in an Animal Model of Obesity.

Today probiotics have been suggested as a treatment for the prevention of NAFLD. Omega-3 fatty acid supplementation may have beneficial effects in regulating hepatic lipid metabolism, adipose tissue function and inflammation. The present study was designed to determine whether probiotics plus omega-3 are superior to probiotics alone on the monosodium glutamate (MSG)-induced NAFLD model in rats. We included 60 rats divided into four groups, 15 animals in each. Rats of group I were intact. Newborn rats of groups II-IV were injected with MSG. The III (Symbiter) group received 2.5 ml/kg of multiprobiotic "Symbiter" containing concentrated biomass of 14 probiotic bacteria genera. The IV (Symbiter-Omega) groups received "Symbiter-Omega" combination of probiotic biomass supplemented with flax and wheat germ oil (250 mg of each, concentration of omega-3 fatty acids 1-5 %). In both interventional groups reduction in total NAS score was observed. Supplementation of alive probiotic mixture with omega-3 fatty acids lead to 20 % higher decrease in steatosis score (0.73 ± 0.11 vs 0.93 ± 0.22, p = 0.848) and reduction by 16.6 % of triglycerides content in liver as compared to probiotic alone. Our study demonstrated more pronounced reduction in hepatic steatosis and hepatic lipid accumulation after treatment with combination of alive probiotics and omega-3 as compared to probiotics alone.

Prevention of NAFLD development in rats with obesity via the improvement of pro/antioxidant state by cerium dioxide nanoparticles.

BACKGROUND: One of the pathogenic mechanisms of the nonalcoholic fatty liver disease (NAFLD) is the accumulation of reactive oxygen species, which in turn aggravates the disease progress. We have investigated novel cerium dioxide nanoparticles (nCeO2) due to their promising antioxidant auto-regenerative ability and low toxicity. METHODS: 30 white male Wistar rats were divided into 3 groups: control, monosodium glutamate (MSG)-induced obesity and MSG treated with nCeO2 (MSG+nCeO2) groups. Newborn rats of control group were injected with saline (control). MSG- and MSG+nCeO2 groups were injected with MSG (4 mg/g concentration, 8 µl/g volume) between the 2nd and the 10th days of life subcutaneously [13]. At the age of 1 month, rats of group II were administered water 2.9 ml/kg orally, MSG+nCeO2 group received 1 mM solution of nCeO2 1 mg/kg orally. 4-months rats were sacrificed and the liver was harvested for histological and biochemical analysis. To assess the morphological changes in the liver we used NAS (NAFLD activity score). The content of lipid peroxidation products and enzymatic activity of superoxide dismutase (SOD) and catalase in the liver were studied by standard biochemical methods [Refs]. RESULTS: In 4-month rats we found significantly lower total score (1.3±0.26 vs 3.6±0.34, p<0.001), degree of steatosis (1.1±0.18 vs 2.1±0.18, p<0.001), manifestation of lobular inflammation (0.2±0.13 vs 1.2±0.2, p<0.001) and ballooning degeneration (0.0±0.0 vs 0.3±0.15, p=0.034) due to NAS in the nCeO2 group compared to the MSG-group. nCeO2 significantly decreased lipid peroxidation in the liver tissue, namely it reduced the conjugated dienes content by 27% (p<0.05), TBA-products - by 43% (p<0.05) and Schiff bases - by 21% (p<0.05). CONCLUSIONS: Due to its antioxidant properties nCeO2 significantly reduces the incidence of NASH and improves the main NAFLD histological features.

Comparative experimental investigation on the efficacy of mono- and multiprobiotic strains in non-alcoholic fatty liver disease prevention.

BACKGROUND: To investigate the efficacy of different probiotic strains, their combinations and forms (alive or lyophilized) in nonalcoholic fatty liver disease (NAFLD) prevention. METHODS: In this study, 70 rats have been used divided into 7 groups of 10 animals in each: I - intact rats, II-VII - rats with monosodium glutamate (MSG)-induced NAFLD. Rats with NAFLD were untreated (group II, MSG-obesity group) and treated with probiotics (groups III-VII). In order to develop NAFLD, newborn rats of groups II-VII were injected with a solution of monosodium glutamate (MSG) (4 mg/g) subcutaneously (s.c.) at 2nd,4th, 6th, 8th,10th postnatal day. The groups III-V received lyophilized monoprobiotics B. animalis VKL, B. animalis VKB, L.casei IMVB-7280, respectively. The group VI received 2.5 ml/kg of an aqueous solution of a mixture of the three probiotic strains (2:1:1 Lactobacillus casei IMVB-7280, Bifidobacterium animalis VKL, Bifidobacterium animalis VKB) at a dose of 50 mg/kg (5 × 10(9) CFU/kg) (g) (intragastrically). The group VII was treated with multiprobiotic "Symbiter" containing biomass of 14 alive probiotic strains (Lactobacillus + Lactococcus (6 × 10(10) CFU/g), Bifidobacterium (1 × 10(10)/g), Propionibacterium (3 × 10(10)/g), Acetobacter (1 × 10(6)/g)) at a dose of 140 mg/kg (1.4 × 10(10) CFU/kg). The treatment with probiotics was started at the age of 1 month. There were 3 courses of treatment, each included 2-week administration and 2-week break. All parameters were measured in 4-month aged rats. RESULTS: Introduction of MSG during the neonatal period leads to the NAFLD development in the 4-months old rats. For steatosis degree there was no significant difference between MSG-obesity group and lyophilized monocomponent probiotics groups (III-V). The highest manifestation of steatosis was observed for B. animalis VKL group (2.0 ± 0.25) as compared to B. animalis VKB (1.70 ± 0.21) and L. casei IMVB-7280 (1.80 ± 0.20). The steatosis score changes between all monoprobiotics groups (III-V) were insignificant. Administration from birth of both alive (VII) and lyophilized (VI) probiotic mixture lead to a significant decrease by 69.5 % (p < 0.001) and 43.5 % (p < 0.025) of steatosis score respectively as compared to the MSG-obesity group (2.3 ± 0.21 %). For both alive and lyophilized probiotic mixtures, reduction of lobular inflammation was observed. These histological data were confirmed by the significant decrease of total lipids and triglycerides content in the liver approximately by 22-25 % in groups treated with probiotic mixtures (VI, VII) compared to the MSG-obesity group. CONCLUSION: We established failure of NAFLD prevention with lyophilized monoprobiotic strains and the efficacy of probiotic mixture with the preference of alive probiotic strains.

Antioxidative effects of cerium dioxide nanoparticles ameliorate age-related male infertility: optimistic results in rats and the review of clinical clues for integrative concept of men health and fertility.

BACKGROUND: Male infertility has largely idiopathic, multifactorial origin. Oxidative stress is a major factor that affects spermatogenesis, in particular in aging. Cerium dioxide nanoparticles (CNPs) due to their antioxidative properties are promising to impact on the development of male infertility. The aims of this study were to investigate the effects of CNPs on fertility parameters in 24-month male rats and to overview relevant literature in the field of personalized treatments, predictive diagnosis, and preventive measures for male health and fertility. METHODS: We included 30 24-month-old male rats. After a week of adaptation to the standard diet, the rats were randomly divided into three groups with ten rats in each. Group 1 (controls) received only a standard diet. The rats of group 2 and 3 in adjunct to the standard diet during 10 days received intragastrically 10 % sodium citrate and citrate-coated CNPs in dose 1 mg/kg, respectively. We assessed sex hormones, epididymal sperm parameters and spermatogenesis, ultrasound, and morphological data of rat reproductive organs. RESULTS: After a 10-day administration of CNPs, we revealed significant decrease of lipid peroxidation product levels in serum and increase of catalase and SOD activity, associated with increase of sperm count (p < 0.001) and improvement in quantitative sperm parameters (motility, viability, and percentage of spermatozoa). We found no significant changes between sperm quantitative parameters in citrate-treated rats and controls and observed age-related decrease of activated Leydig cell number and focal atrophy of the seminiferous tubules. In CNP group, we observed regeneration of seminiferous tubules, increase number and activation of Leydig cells, and 2.5-fold significant increase of serum testosterone. Ultrasound data showed the slight increase of linear measurement and volume of rat testes in CNP group. Review highlights the benefits for predictive diagnosis, preventive measures, and personalized approaches to manage male infertility in the general concept of male health also related to aging. CONCLUSION: Citrate-coated 2-5-nm CNPs lead to increase in sex hormones levels, sperm count, and quality, as well as the activation of spermatogenesis in 24-month-old male rats. Nanoceria demonstrated the perspectives to be an effective infertility treatment via reduction of oxidative stress in male reproductive organs, in particular in aging.

Diagnostic accuracy of acyl-ghrelin and it association with non-alcoholic fatty liver disease in type 2 diabetic patients.

BACKGROUND: Ghrelin is a hormone produced mainly by the cells lining the fundus of the stomach, which is involved in regulation of lipid and glucose metabolism. Two major forms of ghrelin can be found in circulation: an acylated form, and non-acylated form. Serum acyl-ghrelin (AG) concentration is significantly increased in patients with visceral obesity and insulin resistance. This study was conducted to evaluate changes in serum AG levels, its diagnostic accuracy and association with NAFLD in patients with type two diabetes (T2D). METHODS: In this cross-sectional study, 91 T2D patients, age of 40-80 years, were included. All patients were divided into 3 groups. The control group included 28 T2D patients without NAFLD. The main group included 63 T2D patients with NAFLD, which was divided in 2 subgroups depending on transaminase levels: normal (n = 37) and elevated (n = 26) transaminases group. To assess the diagnostic accuracy of AG for NAFLD we used ROC-analysis. RESULTS: We observed 1.5 (p = 0.016) and 2.5 (p < 0.001) fold increasing of serum AG levels in patients with NAFLD and normal or elevated transaminases compared to control groups. In multivariate logistic regression analysis high AG level was an independent, from transaminases activity, triglycerides (OR 1.791; 95 % CI 1.162-2.759; p = 0.008) and degree of IR (OR 1.599; 95 % CI 1.019-2.508; p = 0.044) predictor that associated with NAFLD. When serum AG used as non-invasive marker for NAFLD detection AUROC was 0.835 (95 % CI 0.752-0.918, p < 0.001). The cut-off value was >0.52 ng/ml, with sensitivity, specificity, PPV and NPV - 60.3 %, 92.8 %, 95.0 %, 50.9 % respectively. For distinguishing patients with NAFLD and elevated transaminases from patients with NAFLD and normal values AG was less effective. CONCLUSIONS: Our study has demonstrated that elevated AG level were associated with NAFLD. Patients with elevated transaminases had significantly higher AG levels. An increase of AG over 0.52 ng/ml can be used as a diagnostic marker for NAFLD detection in patients with T2D.