Effect of gold and silver nanoparticles on the morpho-functional state of the epididymis and prostate gland in rats

Metals are widely used in modern medicine: iron, copper, zinc, vanadium, titanium – all of them are vital for treatment of different diseases. Recently a new field of medical technology has emerged, which focuses on the biomedical application of metallic nanoparticles, with a particular interest in a gold and silver-based materials. These structures are already used for photothermal anticancer therapy, drug delivery, bioimaging, radiosensitizers and as drugs themselves. Despite the wide usage of nanoparticles, we still don’t know much about the toxicity of nanomaterials. Nanotoxicological studies are mainly carried out in vitro, but in vivo effects are still elusive. Hence, we focused on the reproductive toxicity of gold and silver nanosized particles. Spherical 10–15 nm gold and silver nanoparticles were synthesized through the reduction of sodium tetrachloroaurate (III) and silver nitrate respectively with ascorbic acid in the presence of sodium polyphosphate as a coating and stabilizing agent. Next, these particles were administered intraperitoneally to the young and adult animals (1- and 6-months old respectively) at 1 mg/kg dose for 10 days. As quantitative markers of functional activity, we used the diameter of epididymal tubules, height and the nuclear cross-section of epididymal epitheliocytes and relative volume of the prostatic epithelium. We showed that intraperitoneal administrations of nanogold to young animals caused no significant histological changes, although we found a decrease in the nuclear cross-sectional area of epididymal epitheliocytes. At the same time, nanogold caused more morphometric changes in adult animals. Similar results were obtained from the nanosilver groups. Silver nanoparticles caused an observable decrease of sperm quantity in the lumen of epididymal tubules with a simultaneous increase in the number of extraepididymal cells in young animals. Morphometric parameters of the epididymis and prostate also decreased. Administration of nanosilver to adult animals also downregulated the morpho-functional state of both organs, although no histological changes were found. We showed that both nanogold and nanosilver can cause adverse effects on the functional activity of the epididymis and prostate of rats. It is worth mentioning that silver nanoparticles were generally more toxic than the gold ones, which correlates with their known mechanism of cellular action. Although the exact mechanisms of gold and silver action require further study, our results are useful for practical usage of nanomaterials in biomedical and clinical practice.

Biochemical parameters in the blood of grass snakes (Natrix natrix) in ecosystems under varying degrees of anthropogenic influence

The grass snake Natrix natrix (Linnaeus, 1758) is a partly hygrophilous species, distributed throughoutUkraine. This snake may be considered as a test object for environmental biomonitoring. Modern biochemical methods make it possible to obtain new scientific data on the effects of anthropogenic pressure on reptiles. Blood is a sensitive and informative indicator of the condition of an organism as it responds quickly to most changes in exogenous and endogenous factors, and reflects negative influences on both individual and, indirectly, populations. Changes in biochemical parameters may be used as biomarkers of the state of health of reptiles in ecosystems under varying degrees of anthropogenic pressure. Due the increase in anthropogenic influence the development and introduction of new methods of perceptual research, collection of up-to-date information and development of a database of reptile biochemical parameters have become an urgent priority. We collected mature individuals of the grass snake in floodplain ecosystems on the right bank of the Dnieper River in Dnipropetrovsk city. Grass snakes from floodplain habitats on the left bank of theSamaraRiver (O.L. Belgard Prysamarskii International Biosphere Station, Novomoskovsk district, Dnipropetrovsk province) were studied as the control specimens. Our study demonstrated statistically significant differences between snakes from the study sites in the amount of albumin, urea and urea nitrogen, and inorganic phosphorus, as well as in alanine aminotransferase (ALT) and alkaline phosphatise (AP) activity. The amount of albumin in the blood serum of specimens from the anthropogenically transformed areas was significantly lower (by 25%) than in that of the snakes caught in the control habitats. Decrease of the albumin concentration usually indicates abnormal processes in the kidneys and liver. According to the changes observed in the concentration of albumin, a corresponding increase in the albumin to globulin ratio by about 30% was found. A statistically significant increase in alkaline phosphatase activity may demonstrate negative physiological changes in tissues of the liver and bones. Increased activities of ALT and AP indicate certain changes in the physiological condition of the liver of snakes from the anthropogenically transformed site. Other studied parameters did not exceed the limits found for different species of reptiles. Significant differences between certain biochemical parameters in the serum of snakes from the studied regions may reflect pathological processes in the grass snakes from transformed ecosystems, mainly in their liver and kidneys. The results can be used in future studies as benchmarks for assessing the condition of the organism of this species in the conditions of anthropogenic stress.

Influence of the liposomal preparation Butaintervite on protein synthesis function in the livers of rats under the influence of carbon tetrachloride poisoning

This article presents the results of research into the influence of the complex liposomal preparation Butaintervit on protein synthesis function in the livers of rats under the influence of carbon tetrachloride poisoning. Intramuscular injection of carbon tetrachloride into rats at a dose of 0.25 ml per100 gof body weight causes antigenic load on the body and leads to disruption of protein synthesis function in the liver. This is shown by reduction in blood levels of total protein and its fractions. Thus, the level of albumin in the serum of rats under the conditions of oxidative stress was 70% lower than in clinically healthy animals. However, the level of total protein in the serum was only 10% lower. This is because, along with the decrease of albumin content in the serum, the levels of globulin protein fraction increased by 8.8%. This has led to albumin/globulin disparities in the serum of sick animals. As a result, the value of A/G coefficient was 0.28 ± 0.03, compared to 0.52 ±0.02 inclinically healthy rats. For the normalization of functional state of the liver under oxidative stress it is advisable to apply the liposomal preparation Butaintervite, which in its structure contains butafosfan, interferon, thistle and vitamins A, D and E. Under conditions of oxidative stress and under the action of the liposomal preparation in the rats from the second experimental group we have found significant increase in the levels of total protein and albumins and a decrease in serum globulin in the animals on the fifth and tenth days of the experiment. On the fourteenth day of the experiment under the conditions of oxidative stress and under the action of the liposomal preparation in the rats from the second experimental group the normalization of protein synthesis function in the liver was observed. The level of indicators of total protein, albumin, globulin and the coefficient of albumin/globulin compared with the control group of animals were within normal values.

Analysis of the application efficiency of the enzyme preparation Ladozim Proxy in the poultry industry

The necessity has been widely noted of improving the nutritional value of forage mixtures used in the poultry industry through the use of enzyme preparations on the basis of phytase for splitting phytic acid and its salts contained in the feed. This technological approach is relevant in feeding farm animals, particularly poultry, as they are unable to synthesise endogenous phytase. On the base of the Agro-Industrial Complex "Oril-Leader" we researched the influence of the enzyme preparation Ladozim Proxy as a feed component for broiler chickens and laying hens on the growth and biochemical parameters of the birds and their egg-laying capacity. The feed ration consisted of wheat, corn, sunflower meal, extruded soy and protein-vitamin complex Biomix. For the research experimental and control groups of broiler chickens were formed (50 birds in each group) and laying hens (20 birds in each group). The experimental feeding period was 35 days for the chickens and 30 days for the hens. The research used physical-chemical methods for determining the content of nitrogen, calcium and phosphorus in the feed, the blood and the excrement of the birds. Statistical data processing was carried out by ANOVA. It was established that the use of Ladozim Proxy in the amount of 60–100 mg/kg feed promotes an increase of the live weight of chickens by 21%. The findings can be explained by an increase of the feed nutrients available for digestion in the gastrointestinal tracts of the poultry due to the hydrolytic action of the enzyme complex added to the phytic-containing components of the feed. The result of these phenomena is also a positive dynamic in assimilation of nitrogen, calcium and phosphorus in the feed by chickens of the experimental group: use of these elements increased by 10–16%. The preparation had no effect on hematological indices of the organisms of the poultry and increased the amount of calcium and phosphorus in the blood. The use of the preparation has not only physiological, but also economic importance because it allows a reduction in feed intake and expenditure on feed for poultry reared for agricultural production. The use of the phytase preparation does not affect the quantitative indicators of egg-laying by hens, but an increase in the thickness of the egg shells was noted, due to deposition of calcium in the organism of the experimental group.

Pancreatic response of spontaneously hypertensive rats to the exogenous administration of melatonin in the spring and autumn

The aim of the study was to investigate and compare the effect of exogenous melatonin administration on the morphofunctional state of the exocrine and endocrine parts of the pancreases of spontaneously-hypertensive rats (SHR line) in spring and autumn. The research was conducted on 48 male SHR line rats, age 4 months. The rats of the experimental group received melatonin (Unipharm Inc.,USA) every day (at 10 am) orally at a dose of 5 mg/kg of body weight. The duration of the experiment was 28 days. Morphofunctional activity of the exocrine and endocrine parts of the pancreas was assessed using morphological and morphometric methods. The histological preparations were made by the standard methods. The slides were photographed using a digital camera on a Nikon microscope. The morphometric analysis was performed on digital images using the computer program Image J. The morphological signs of the exocrine pancreas activity were greater in the control rats in spring compared to autumn. Conversely, the activity of the endocrine part of the pancreas in control animals was higher in autumn, than in spring. Exocrine pancreas activity increases after administration of melatonin regardless of time of the year. This was evidenced by the increase in size and height of the epithelial acini, and the number of exocrinocytes placed in them. The width of the interlobular and interacinus connective tissue layers in the pancreas decreased in spring (mostly) and in autumn. The activity of the endocrine part of the pancreas increased after administration of melatonin in spring. Proof of this was an increase in the number and size of islets of Langerhans and the number of endocrinocytes they contained, compared with control values. The number and size of islets of Langerhans, the number and density of the endocrinocytes they contained decreased in the rats which received melatonin in autumn. This indicates a reduction of the endocrine pancreas activity. These results may have implications for therapeutic dosing of melatonin in patients with hypertension in different seasons of the year.

Phagocytic and metabolic activity of neutrophils of rats in the early stages of post-embryonic development under the influence of biologically active substances of the salt extract of Hirudo verbana

The nitroblue tetrazolium test reflects the degree of activation of oxygen-dependent mechanisms of bactericidal activity of phagocytotic cells, so the increase in nbt-test and phagocytic indices shows increased activation in the bacterial process and enhanced protection with the help of the absorption ability of the cells in the body. We examined females of nonlinear rats after feeding of their offspring, which 2 weeks before and 2 weeks after mating were injected intraperitoneally with antigens of saline extract of medical leeches (4 injections) and the offspring at days 1, 15, 30, 45, 60. The dosage of salt extract antigens was carried out by determining the protein concentrations using the Lowry protein assay. The animals were immobilized using an immobilizing device. The animals were distributed into three groups: the first research and experimental group of animals was subject to the influence of medical leech salt extract antigens to the amount of 0.5 ml (at the rate of 3 µg/g of animal weight); the second group of intact animals was not subject to intervention, the third control group of animals was administered intraperitoneally saline 0.5 ml of 9% saline solution. We studied females after feeding offspring and offspring at 1, 15, 30, 60 days. The animals were kept in vivarium conditions in individual cages on a standard diet. In the experiment we used 60 females of non-linear rats and 300 of their offspring. All animals were decapitated under ether anesthesia. After that, we took blood, made a solution with crystal heparin and examined the laboratory parameters: total number of leukocytes and leukocyte formula. Evaluation of nonspecific resistance of the organism was investigated using tests which characterize the activity of neutrophils (nbt-test, phagocytosis indices). The results of our studies revealed a positive effect of antigens of saline extract of medical leeches on the absorptive and metabolic activity of neutrophils in the females and their offspring in all study periods compared with the control, an increased number of leukocytes, due to the change of the process of differentiation and deposition of cells in immunogenic organs on proliferation. Changes in immunological parameters indicate the immunostimulatory effect of AG ML on leukopoiesis and on the functional activity of neutrophils – the first line of defense against entry into the body of a variety of bacteria, fungi and protozoa.

Relationships of oxidative stress and systemic inflammation markers depending on the degree and duration of hypertension

Arterial hypertension (AH) is a heterogenic and multisystem disease. It has been suggested that oxidative stress (OS) and systemic non-specific inflammation may be involved in pathogenesis of cardiovascular pathology including AH. The aim of our study was to characterize the plasma C-reactive protein (CRP) level as a marker of systemic inflammation in relation to OS development (on the base of 8-isoprostane level assessment), depending on duration and degree of AH. We examined 117 persons, of which 102 patients from 30 to 65 years old (average age – 54.7 years) who had previously not been receiving regular antihypertensive therapy had I–III degrees of essential hypertension and 15 healthy persons (average age – 48.7 years). In 34 patients from this group the degree of OS activity was determined by 8-isoprostane level as the main marker of OS. The control group consisted of 10 healthy persons, by age and gender comparable with the study group. Determination of plasmatic CRP levels and the level of 8-isoprostane in the serum was performed by ELISA. The study established an increase of the plasmatic CRP levels in patients with hypertension, and a statistically significant increase of serum 8-isoprostane content in hypertensive patients compared to the control group. When assessing the relationship of 8-isoprostane and CRP content in patients with different degrees of hypertension we found that the strongest positive relationship between their levels was observed in the case of I degree hypertension. This may indicate the role of oxidative stress in the pathogenesis of hypertension as a damaging mechanism which contributes to the activation of immune mechanisms and further progression of the disease. Increased CRP and 8-isoprostane levels confirm the involvement of autoimmune mechanisms and oxidative stress in the pathogenesis of hypertension. The level of C-reactive protein is dependent on the duration of hypertension, while the 8-isoprostane levels – only on degree of hypertension. A raised level of C-reactive protein can be used as an independent marker of systemic inflammation in patients with arterial hypertension.

Modern diagnostics of nonalcoholic fatty liver disease: noninvasive research methods

Nonalcoholic fatty liver disease (NAFLD) is increasingly recognized as the most common cause of chronic liver disease worldwide. NAFLD represents a wide spectrum of conditions, ranging from simple benign steatosis to nonalcoholic steatohepatitis, which sometimes progresses to cirrhosis and hepatocellular carcinoma. The pivotal issue in the management of patients with NAFLD is the diagnosis of steatohepatitis and fibrosis at an early stage. In this review we present recent data on nonalcoholic fatty liver disease evaluation. Although liver biopsy is regarded as the gold standard for assessment of hepatic steatosis and steatohepatitis, its use has several limitations, including the potential risk of sampling errors, intra- and interobserver variability, invasiveness and the stress it causes to patients, the high cost and the potential for complications. In this review a simple and reliable non-invasive alternative with indicated sensitivity and specificity is described. Non-invasive markers should aim ; in primary care settings, to identify the risk of developing NAFLD among individuals with increased metabolic risk; in secondary and tertiary care settings, to identify those with a worse prognosis, e.g. severe steatohepatitis; monitor disease progression; predict response to therapeutic interventions. Achieving these objectives could reduce the need for liver biopsy. Thus, according to the natural history of NAFLD, all patients with a low risk of developing advanced disease, eventually diagnosed by one of above non-invasive parameters, could be referred to primary care, whereas subjects at high risk of developing advanced disease should be sent to specialists for the evaluation of the degree of fibrosis and the choice of specific management. According to the Clinical Practice Guidelines for the management of NAFLD, ultrasound is the preferred first-line diagnostic procedure for imaging of NAFLD, as it provides additional diagnostic information. Whenever imaging tools are not available or feasible (e.g. large epidemiological studies), serum biomarkers and scores are an acceptable alternative for the diagnosis of steatosis. The combination of biomarkers/scores and transient elastography might confer additional diagnostic accuracy. At the same time, the identification of advanced fibrosis or cirrhosis by serum biomarkers/scores and/or elastography is less accurate and needs to be confirmed by liver biopsy, according to the clinical context. Аlthough there are a range of controversial issues regarding the use of non-invasive methods for the assessment of NAFLD, these methods are being actively developed, researched and introduced into clinical practice as an equivalent and substitute for liver biopsy.

Association of group erythrocyte antigens and B-cell non-Hodgkin lymphoma

We analyzed the frequency distribution of blood groups AB0, Rhesus and Kell in patients with B-cell non-Hodgkin lymphoma. An increase of D− patients (31.0%) among ill in comparison to healthy individuals (16.5%) was found. Based on published data, we exclude weakening or alteration of the D antigen for this pathology. We found an association of B-cell non-Hodgkin lymphoma and D− phenotype. It is established that the chances of developing the disease in D− individuals is 2 times higher than for D+ individuals. However, we did not find a significant association of frequency of minor antigens Rhesus (C, c, E, e, CW), K antigen Kell system, A and B antigens AB0 system and the development of B-cell non-Hodgkin lymphoma. We have analyzed the statistics of incidence of non-Hodgkin lymphoma inWestern Europeand Asian countries. We noticed a roughly twofold increase in the incidence of non-Hodgkin lymphoma inWestern Europecompared to Asian countries, where the proportion of D− individuals is almost zero. Parallel with the results of our study, it has been found that the chances of D– persons developing B-cell non-Hodgkin lymphoma are twice as high as for D+ individuals. InUkraine,BelarusandRussiathe incidence of non-Hodgkin lymphoma is almost at the level of Asian countries even though the percentage of D– individuals corresponds to Western European countries (15–16%). This can be explained by other genetic factors, or reduction in quality of care and diagnostics in post-Soviet countries compared toWestern Europe. Thus, D antigen may have a protective role in the morbidity of non-Hodgkin lymphoma.

Dyslipidemia in patients with arterial hypertension and extrasystoles

The significant incidence of hypertensive disease among the Ukrainian population makes the search for new methods of management and treatment of patients suffering from this pathology a matter of high priority. Our research is focused on study of changes of some biochemical parameters in patients suffering from hypertensive disease as well as patients suffering from hypertensive disease and coexistent extrasystoles. Arterial hypertension leads to different structural and functional changes in the cardiovascular system. In many patients these changes may result in rhythm disorders. 120 patients have been examined (42 men and 78 women) between the ages of 27 to 81 (average 59.8) suffering from hypertensive disease of the II stage as defined by the Ukrainian Association of Cardiologists and from extrasystoles (more than 30 episodes per hour of research) of different topography and 30 persons (13 men and 17 women) suffering from hypertensive disease of the II stage between the ages 30 to 76 (average 56.4) without rhythm disturbance). The first clinical group was formed by 54 (45%) patients suffering from repeated supraventricular extrasystoles (SE) (average 62 ± 2 SE per hour of research). The second clinical group was formed by 42 (35%) patients suffering from repeated ventricular arhythmia (VA) (average 37 ± 2 VA per hour of research). The third group was formed by 24 (20%) patients suffering from both repeated SE and VA (average 48 ± 3 SE and 32 ± 3 VA per hour of research). We conducted 24 hours’ monitoring of blood pressure and electrocardiography using the “Dia Card”-system (“Solvaig”, Ukraine) for all patients. This revealed that ventricle extrasystoles and combined extrasystoles in patients with essential hypertension, II stage are associated with more unfavourable changes in 24 hours’ profile of blood pressure. The paper analyses the cardiac arhythmia structure of heart rate variability in patients with arterial hypertension (AH) and extrasystoles. The findings confirmed the presence of desynchronization in the activity of the neurohnmoral system. Thus, different mechanisms of arhythmogenesis were revealed in patients with arterial hypertension. In the results, frequent ventricular and combined extrasystoles were associated with an unfavourable blood pressure profile during 24-hours monitoring as well as higher end-diastolic left ventricular volume, diastolic dysfunction, aortic valve fibrosis, aorta remodeling, endothelial dysfunction and more prominent lipid disorders. Supraventricular extrasystoles were associated with hypertriglyceridemia, left ventricular diastolic dysfunction and left atrium enlargement.