Melanin synthesis by black yeast-like fungi Psedonadsoniella brunnea: dependence of L-tyrosine quantity in the cultural medium

To determine the optimal conditions for the synthesis of melanin by black yeast fungi Pseudonadsoniella brunnea (Basidiomycota, Agaricomycotina, Agaricomycetes, Polyporales, Meripilaceae), depending on the amount of L-tyrosine in the culture medium was the purpose of the work. The standard Malt Extract Broth (MEB) liquid nutrient medium was used within this study. L-tyrosine was added to the culture medium in a quantity of 0.01, 0.025 and 0.05%.To obtain the melanin the cultivation of Pseudonadsoniella brunnea was carried out at pH 1-1.5, temperature + 21 ± 1 ° C during 7 days. Statistical processing of the results was carried out using generally accepted methods of variation statistics. It has been established that the level of melanin synthesis by black yeast-like fungi Pseudonadsoniella brunnea depends on the amount of L-tyrosine introduced into the culture medium. The MEB nutrient medium containing 0.05% L-tyrosine in this series of experiments found to be the best composition for obtaining melanin by the strain-producer Pseudonadsoniella brunnea. Compared to control (MEB without L-tyrosine), the amount of melanin synthesized by Ps. brunnea in these conditions increased by 2.5 times. The further research into the optimal conditions for the cultivation of black yeast-like fungi Pseudonadsoniella brunnea in order to obtain melanin is relevant and promising.

The content of matrix metalloproteinases in bladder cancer tumors

Bladder cancer (BC) is the 9th in frequency oncologic disease, with the highest number of patients in developed countries. The disease is more commonly diagnosed in men – about 75%. The ability to invade surrounding tissues and metastasis to individual organs is one of the fundamental properties of malignant tumors. According to modern ideas about mechanisms of metastasis, basement membranes and extracellular matrix are the main barriers, and tissue structures are needed to overcome the invasive growth of tumor cells. Almost all of the extracellular matrix components can be destroyed by the use of metal-matrix proteinases (MMP), moreover, the precursors of growth factors and adhesion molecules on the cell surface may be the MMP substrates. MMPs also participate in epithelial-mesenchymal transformation, which provides metastasis. Previous studies by other authors point to an increase in the biosynthesis of various MMPs in the outbreak of tumors and in metastases, such changes associated with the degree of differentiation of the tumor, the depth of the invasion, as well as their association with poor further prognosis and low survival rates in patients with various cancers. The purpose of our work was to investigate the content of MMP-1, 2, 3, 8 and TIMP-1 in the tumors and walls of the healthy bladder in patients with BC, depending on the stage of TNM classification. We investigated the increase in the content of MMP-1, 2, 3, 8 and TIMP-1 in samples of healthy bladder walls for stage 3 and 4 of the BC, indicating the participation of the investigated parameters in the growth and invasion of bladder tumors. The results of our study are consistent with the results of previous studies conducted in the study of other neoplasms, which indicate the relationship between the studied parameters with the development of cancer.

Eeffect of an aquaeous extract of Phaseolus vulgaris pods on morphological properties of post-burn esophagus

The most widespread chemical damage in children – burn of the esophagus. It occurs when accidentally swallowing substances that have in their composition alkali, acid or other caustic substances. The depth of burn depends on the concentration and time of contact of the mucous membrane with the affecting agent. As a result of penetration of alkalis and contact with the wall of the esophagus, along with necrosis, damage to the mucous membrane, submucosal and muscular membrane of the esophagus occurs. Despite a large number of drugs that have been tested in the treatment of complications caused by esophageal burns, there is little information that would indicate the high effectiveness of the previously used drugs. The analysis of literature data shows the positive influence of polyphenols of natural origin on the normalization of physiological and biochemical parameters in various pathologies. The extract of common beans (Phaseolus vulgaris pods extract (PVPE)) is an example of a rich bioactive component of the source with beneficial characteristics for humans. Therefore, the purpose of our work was to investigate the effect of the Phaseolus vulgaris pods extract on the healing of post-burns wounds. When morpho-functional characteristics of the esophagus changes in the conditions of alkaline burn of the esophagus of the 2nd degree, it was shown that at 1 day of the experiment there is a violation of the mucous membrane, which manifests itself in the destruction of the multilayered epithelium, namely, in the submucous membrane there is a significant accumulation of leukocytes, leukocytic infiltration is fixed in the partly destroyed muscle layer. At 31 days of study in a group of animals with burn of the esophagus there is a certain restoration of the integrity of the mucous membrane of the esophagus, mainly with hyperplasia; there is also a decrease in the presence of leukocytes; however, there is an enlargement of the connective tissue in the submucosal membrane, which may indicate the formation of a scar. The data of the morphological description are confirmed by the morphometric parameters of the calculation of the stenosis index. With the use of extracts of beans under chemical burns esophagus conditions, improvement of the morpho-functional status of the esophagus of rats was noted: the formed connective tissue scar does not significantly affect the stenosis index (its significance is the intermediate position); the high level of leukocyte infiltration is reduced to the control parameters, which may indicate the obvious therapeutic effect of the extract of beans pods at alkaline burns of the esophagus.

Biological value of raspberry fruits

The human body needs a constant flow of biologically active substances. Berry products, including raspberries, are a source of natural and affordable BAS. Raspberry berries contain a complex of BAS, in particular fructose, glucose, organic acids, vitamins A, B1, B2, C, P, E, essential oils, pectins, fiber, complex of polyphenols, copper salt, iron, potassium, folic acid, etc. Berries are salicylic acid and can for a long time maintain the beneficial properties of various processed products. Berries contain a lot of food fibers, it is beneficial for digestion. Berries contain betasitosterol and anthocyanins have anti-sclerotic properties, and coumarins normalize blood coagulation. Berries contain a lot of food fibers, it is beneficial for digestion. Raspberry berries contain coarse dietary fiber, β-sitosterol, anthocyanins and coumarins, these substances have a positive effect on digestion, have anti-sclerotic properties and normalize blood clotting. The purpose of the research aimed at carrying out a comparative characterization of the basic biochemical substances content and the quality of various berries and raspberry hybrids grown at the research field "Fruit and Vegetable Garden" of NULES of Ukraine. During the research the complex of international and state regulatory documents were used, in particular: the berries were selected in the state of technical maturity, in accordance with the state standard 7179: 2010; the sample size corresponded to ISO 874-2002; the determination of the components of the biochemical composition was carried out: the content of dry soluble substances were determined according to ISO 2173-2013, invert sugars – state standard 4954: 2008, titrated acids – state standard 4957: 2008, ascorbic acid – ISO 6557-2: 2014; organoleptic evaluation – according to the generally accepted methodology. Based on the obtained results, it was found that among the studied samples of berry varieties Babine Lito, Sontse Kieva, hybrids – Hybrid №4a, Hybrid №2 were characterized by the highest organoleptic and biochemical parameters, therefore it is expedient to use them both for consumption in a fresh form and for processing. The obtained results should be taken into account during the planning of raspberry planting and in further selective work during the creation of competitive varieties and hybrids of raspberries with a high content of biologically active substances in berries.

Isolation and characterization of the mutant N-terminal catalytical module of the B. taurus Tyrosyl-tRNA synthetase with the replacement of Trp87 and Trp283 by alanine

Aminoacyl-tRNA synthetase is one of the major enzymes of protein synthesis. The mammalian tyrosyl-tRNA synthetase consists of two structural units, the N-terminal catalytic (mini TyrRS) and the C-terminal cytokine-like modules. In a full length TyrRS, the N-terminal module carries out the catalytic function of binding the amino acid to tRNA, while the C-module adjusts and stabilizes the placement of tRNA in the active center of the enzyme. After cleavage of tyrosyl-tRNA synthetase with elastase on the mini TyrRS and C-module, the latter exhibit cytokine properties. The aim of the work was to optimize the expression of cloned cDNA miniTyrRS Bos taurus in plasmid pET30a-39KYRS in which the tryptophan codons at position 87 and 283 are replaced with alanine codons using the site-directed mutagenesis, and to obtain the mutant one-tryptophan protein of the mini BtTyrRS for further study on using methods of fluorescence spectroscopy of conformational changes of the enzyme at the stage of tyrosyladenylate formation and in interaction with the acceptor end of tRNATyr, as well as determination of the effect of tryptophan residus in positions 87 and 283 in its structure on the structurally dynamic and functional properties of the enzyme. It was found that the replacement of two tryptophan codons into the alanine codons in the cDNA of the mini TyrRS cloned in the expressing plasmid pET30a-39KYRSW40 does not affect the synthesis and solubility of the mutant form of the enzyme in the strain E.coli BL21 (DE3) pLysE. The amount of soluble form of the recombinant mutant mini BtTyrRS in the cytoplasm of bacterial cells, when expressed in E. coli BL21 (DE3) pLysE strain, is significantly enhanced by incubation of bacterial culture at a temperature 25 ° C compared to a culture incubation at 37° C. The yield of the obtained purified protein of the mutant mini BtTyrRS is 2.5 mg per average from 100 ml of culture medium, which is sufficient for further structural and functional studies of the mutant form of the enzyme. The compact structure of the recombinant protein is shown by fluorescence spectroscopy.

Oxidative modification of proteins in rat serum under experimental osteoarthrosis and long-term administration of a multiprobiotic

The aim of the study was to investigate the effect of multiprobiotics on the content of products of oxidative modification of proteins and the level of sulfhydryl groups in blood serum of rats during monoiodoacetate-induced osteoarthritis. The study was carried out on white non-linear, sexually mature male rats (weight 180-240 g), according to general ethical principles of experiments on animals. All animals were divided into four experimental groups. The first group - Control: animals got injection into knee ligament 0.05 ml of 0.9% NaCl solution on the first day of the experiment and then got intragastric administration 1 ml of drinking water per 1 kg of the animal weight daily for 14 days from the 8th to 22nd days. The second group - Multiprobiotic: animals got injection into knee ligament 0.05 ml of 0.9% NaCl solution on the first day of the experiment and then got intragastric administration 140 mg/kg of multiprobiotic Symbiter® (Prolisok ", Ukraine) diluted in 1 ml of drinking water per 1 kg of animal weight. The third group, MIA-induced OA: animals got injection into knee ligament 1 mg of sodium monoiodacetate, dissolved in 0.05 ml of 0.9% NaCl on the first day of the experiment and then got intragastric administration 1 ml of drinking water per 1 kg of the animal weight daily for 14 days from the 8th to 22nd days. The fourth group – MIA-induced OA + Multiprobiotic: animals got injection into knee ligament 0.05 ml of 1 mg of sodium monoiodacetate, dissolved in 0.05 ml of 0.9 % NaCl on the first day of the experiment and then got intragastric administration 140 mg/kg of multiprobiotic diluted in 1 ml of drinking water per 1 kg of animal weight. All animals were killed on day 30 of the experiment, according to the protocol of the ethics committee with rapid blood sampling. The content of the products of oxidative modification of proteins (OMP) and oligopeptides was determined by the level of carbonyl derivatives that were detected in reaction with 2,4-initrophenylhydrazine. The level of total, protein-bound and non-protein sulfhydryl (SH) -groups was measured by the Elman method. It has been established that MIA-induced OA disturbed oxidative-antioxidant balance of the rat serum: the content of the products of oxidative modification of proteins increases and the content of sulfhydryl groups decreases in the serum. It was shown that with the long-term administration of multiprobiotics in animals with MIA-induced OA, the above indicators were restored.

The effects of melatonin administration in determined times of day on the kidney in rats with high-calorie diet-induced obesity

Kidneys, like the cardiovascular system, are one of the main target organs, the most vulnerable to obesity, because the first take on the metabolism correction function at an increasing excess of fat tissue in the body. Kidney affection under obesity is a multifactorial thing that is caused by a number of processes, including inflammation, oxidative stress, lipid metabolism disorders, renin-angiotensin-aldosterone system activation, insulin resistance and other factors. An optimal candidate for reducing the harmful effects of kidneys obesity should be a compound that simultaneously exhibits anti-inflammatory and antioxidant properties, controls the circadian rhythm, and also affects on the adipokines secretion. The molecule that meets these conditions is melatonin.The aim of our study was to determine morpho-functional state (morphology characteristic of kidney glomeruli and tubules; morphometric parameters: area and density of glomeruli) of kidney in rats with high-calorie (high fat) diet-induced obesity after melatonin administration in determined time of the day. Melatonin was administered daily by gavage for 7 weeks in dose 30 mg/kg 1 h before lights-off (ZT11) rats with high-calorie diet (HCD). Rats with HCD had huge changes in kidney morphology, which manifested in presence of numerous mesangial cells outside glomeruli and lipid droplets in tubules epithelial cells, while area and density of glomeruli in cm2 decrease. In general kidney with above mentioned characteristic from HCD rats lose their ability to conduct strongly renal function. After melatonin used in rats with HCD arise leveling of pathological changes, which associated with consumption of HCD. Namely, in rats with development obesity melatonin administrations led to increase area and density in comparison to HCD group, moreover glomeruli density reach control values. This is suggest that melatonin have protecting effect against glomerular degeneration. In conclusions, melatonin influence on kidney morpho-functional state in rats with HCD and turn back pathological its changes, moreover evening administration can use for obesity therapy via its strong action on conservation glomerular morphology.

Relation of success in study of foreign language with speed of interhemispheric transfer of information

Bilingual speakers seem to outreach monolingual speakers in performing non-verbal tasks for testing executive functions, such as Simon, Flanker and Stroop tasks, as well as in capacity of working memory. Other researchers have doubts about these cognitive benefits of bilinguals and multilinguals. The study used a combined test with the tasks of the Stroop, Poffenberger, Sperry in native (Ukrainian) and foreign (English) languages. Schoolboys of lyceum were the subjects surveyed (14-15 years old). Stimuli (the word "Green" or "Red", "Blue" or "Yellow" written in relevant or irrelevant color) were exposed on the right or left from the center of the screen. In the case of congruence the word and its semantic meaning should press one button by the ipsilateral hand ("yes"), while in the case of mismatch – the other button by the contralateral one ("no"). Latent periods of response to stimuli, which reflect the speed of the interhemispheric transfer of information, were taken into account. Correlation analysis of the success in study in the nine subjects of the three blocks (humanities, natural and formal disciplines) reveal a direct correlation of speed of reaction when performing the complex Stroop test in both the native and English languages with the success in the learning English language, what may indicate on special dependence of the success from interhemispheric interaction. In order for the foreign language to be automated and become "all the more native", it is need the fast access of the executive structures of the brain, such as the front-parietal neural network, to the linguistic neural networks, presented in both hemispheres. According to literature, the inhibitory control mechanism from the dorsolateral prefrontal cortex as the key structure of the front-parietal brain system may be one of several mechanisms underlying bilingual superiority. The results obtained by us complement this conception, indicating the importance of the speed of interhemispheric interaction.

C-reactive protein and medium weight molecules concentration in serum of rats under conditions of carrageenan-induced inflammation and prolonged prophylactic administration of chondroitin sulfate

The aim of the work was to investigate the preventive effect of Chondroitin Sulfate on the concentration of C-reactive protein and medium weight molecules in blood serum of rats at local inflammation of the hind limb. The studies were conducted on white non-linear, sexually mature male rats weighing 180–240 g, in compliance with the general ethical principles of experiments on animals. All animals were divided into four experimental groups. The first group – control: animals sub-planar injected 0.1 ml of 0.9 % NaCl solution into the posterior right limb. The second group – animals received a therapeutic dose of 3 mg x kg-1 chondroitin sulfate daily for 28 days daily. The third group – animals were infused intramuscularly with 0.1 ml of 0.9 % NaCl solution in the posterior right limb for 28 days and for 29 days inflammatory edema of the limb was stimulated (animals were sub-planar injected with 0.1 ml of 1 % carrageenan solution to the posterior right limb ) The fourth group – for 28 days rats were daily intramuscularly injected with a therapeutic dose of 3 mg x kg-1 chondroitin sulfate, after which on 29th day, inflammatory edema of the limb was stimulated. Animals were killed 3 hours after injection of carrageenan solution according to the protocol of the ethical committee, and then blood sampling for further research was quickly taken. The total number of animals involved in experimental studies was 40 individuals. Creactive protein concentration was determined by turbidimetric method. The content of medium weight molecules was determined by screening method. It has been established that with carrageenan-induced inflammation of the posterior limb, the concentration of C-reactive protein and medium weight molecules increases in the serum. It was shown that the prophylactic administration of chondroitin sulfate based drug on animals with carrageenan-induced inflammation restored the abovementioned parameters.

Morphological features of the mucous membrane of the colon of patients with early and late ulcerative colitis debut

The purpose of the research was to determine the morphological features of the mucous membrane of the large intestine of patients with UC depending on the age of patients. A comparative analysis of the incidence and type of UC of patients with early onset and late debut of the disease was performed. It has been stated that for patients with a debilitating post-traumatic disorder, the left ventricular and total forms are characteristic; in patients with late debut, the distal form of UC predominates. The main trends in the activity of UC in patients with early onset and late debut have been determined. In the analysis of the degree of activity of UC revealed that in patients from 50 years of age and older, with the late debut of the disease, the first one is mostly found to be the least degree of activity, the second – the middle and the third – the high degree, they are found to be less likely. In turn, in patients under 50, the second and third stages of the disease are most often noted. Another component of the main characteristics of the UC was the histological changes of the mucous membrane of the large intestine. Thus, the conducted studies allowed to state that the number of PAS-positive substances in mucus is significantly higher in patients with distal and left-sided UC. In patients with UC there is a marked mucosal intestinal mucosa of qualitative and quantitative composition of mucus. These changes were characterized by a decrease in MUC2 as the disease progressed, as well as levels of MUC4 and TFF3. In patients with early onset of NSC, there is a more intense reduction in the number of mucins and TFF3 already with a minimal activity of UC associated with low levels of mucin from the intracellular mucus and indicates a more aggressive course of it. At the same time, patients with a late debut revealed a sharp decrease in the number of MUC2 and TFF3 with high activity of UC.