Issue 3 | September 2023
Over the past 20 years, significant advances have been made in the understanding of cell and tissue culture in the fields of bioengineering, cell biology, and genetics; however, these achievements have largely focused on mammalian systems. In vitro studies on fish cells and tissues have been relatively limited, but the use of fish cell lines as in vitro models for environmental toxicology, particularly cytotoxicity analysis, has been significant. In addition, cultured cells have been used to study fish parasites and as in vitro models for screening immunostimulants. Skeletal muscle is the most important edible tissue in fish and rapid muscle growth determines the advantages of in vivo aquaculture. The generation of fish muscle cell lines can help to provide a reliable platform for deciphering the mechanism of fish skeletal muscle growth both in vivo and in vitro. In addition, cultured fish meat is a promising technology for animal protein-based foods and the concept of cell biomass meat from fish needs to be further developed. Our data demonstrated the feasibility of obtaining and culturing sterlet muscle cells in vitro. This study highlights the potential advantages of cell aquaculture over traditional fishing and aquaculture, and the potential applications of fish muscle cell lines in the study of fish skeletal muscle growth and the production of edible cultured fish meat products.
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This study evaluated the effect of different diets on the probiotic (lacto-, bifidobacteria) and opportunistic (yeast, Escherichia coli) intestinal microflora of CD1 mice. The high-fat diet contained 40% animal fat (lard) and the high-fiber diet contained 40% freeze-dried fiber. The intestinal microflora was determined by the standard method of seeding the contents of the intestine on selective culture media (MPС, Blaurock, Sabouraud, Endo). The results showed that on the 50th day of the experiment in the group of mice with a high fat content, the population of probiotic cultures of lacto- and bifidobacteria decreased, while the population of yeast and enterobacteria increased, compared with the starting point of the experiment and the control group of mice. The weight of mice in this group by the end of the experiment increased by 16%. In the group of mice with a high content of insoluble fiber, a decrease in the populations of probiotic cultures, yeasts and enterobacteria was observed. At the same time, the weight of mice increased by 13.6%. Thus, high fat intake in the diet entails possible disturbances in the intestinal microbiota, an increase in opportunistic microflora, which can lead to intestinal diseases. When using a large amount of insoluble fiber, on the contrary, it leads to a decrease in microflora in general. This is most likely due to a lack of nutrients and enough nutrients (proteins and fats) in the diet, which are still necessary for the microflora.
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Norovirus infection is a leading cause of non-bacterial acute gastroenteritis. This study aimed to analyze the antigenic properties of the VP1 protein of norovirus GII.4 Sydney [P16] circulating in Russia. The analysis was conducted using in silico methods. VP1 amino acid sequence data was used to identify T-helper and T-killer epitopes, linear and conformational B-cell epitopes to assess the conservation of epitopes, and allergenicity of VP1. T cell epitopes with the highest estimated immunogenicity were identified at positions 207-223 and 378-394 in the S- and P-domains of the protein. The tertiary structure of VP1 was modeled, and 2 linear and 47 conformational B-cell epitopes were identified. In addition to the previously described epitopes, a new putative B-cell epitope was identified at position 307-316 of the P2 subdomain. In silico analysis of the primary and tertiary structure of the norovirus VP1 protein showed that it is not allergenic and has various immunogenic epitopes, potentially capable of inducing T- and B-cell immune responses.
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The purpose of the study was to study the fronto-occipital organization of cerebral circulation during short-term mental stress, as well as to study the relationship between the spatial organization of cerebral circulation during mental stress and the effectiveness of behavior control. In this work, attention was paid to potential typological characteristic – fronto-occipital asymmetry of cerebral circulation. Known data suggest a relationship between changes in the spatial distribution of cerebral circulation and the productivity of mental activity and the effectiveness of behavior control. Using the method of rheoencephalography, 40 students (aged 19–23 years) studied the parameters of cerebral circulation during mental stress (GO/NOGO test). The stimulus material was presented on a monitor using the PsychoPy program. The number of errors was estimated. The rheoencephalogram was recorded using the “Reo-Spectrum” device. The spatial organization of pulse blood supply in the basin of the internal carotid and vertebral arteries in the initial state and during mental stress was assessed by the fronto-occipital gradient. With a short-term mental load, students showed an increase in both general and particular fronto-occipital gradients. A negative correlation was established between the increase in the right-diagonal fronto-occipital gradient (reactive control axis) in the first minute of the mental load and the number of errors in the GO/NOGO test. The prospects of using the fronto-occipital organization of cerebral circulation to predict the effectiveness of mental activity are discussed.
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The Influence of BDNF on Anhedonic Behavior in an In Vivo Model of Chronic Unpredictable Mild Stress
Depression is a significant global medico-societal concern. The serotonin system plays a pivotal role in modulating responses to acute stress and is implicated in the development of depressive and anxiety disorders. Recent research has increasingly focused on the potentially beneficial impacts of activating previously less-studied 5-HT4R and 5-HT7R subtypes on cognitive functions in the context of anxiety and depression. Additionally, intercellular adhesion molecules have been associated with the structural remodeling of neurons related to stress and mood disorders, potentially establishing functional connections with serotonin receptors. Furthermore, it is established that the exogenous administration of the neurotrophic factor BDNF can ameliorate the functioning of serotonergic neurons in the brains of rodents. This study aimed to investigate the influence of exogenously administered BDNF on the expression of 5-HT4R, 5-HT7R, and CD44 during a depressive-like state induced by chronic unpredictable mild stress (CUMS) in C57Bl/6 mice. The findings demonstrated that intranasal BDNF administration at a dose of 0.4 μg/kg for seven days sustained normal sucrose preference levels in animals following 21 days of CUMS exposure. While BDNF treatment did not impact the CUMS-induced reduction in mRNA expression of 5-HT4R and 5-HT7R across examined brain regions (cortex, hippocampus, and cerebellum), it did prevent the decrease in CD44 and TrkB receptor expression levels in the hippocampus. Additionally, it maintained BDNF expression levels in the cortex, although not in other brain regions. These results suggest that the application of BDNF in CUMS models has an antidepressant effect without directly affecting serotonin receptors, but probably by modulating 5-HT7R-CD44 interactions.
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In 2010, the first scientific study was published on genome-wide data on ancient DNA belonging to a male of the Paleo-Eskimo culture, who lived about 4000 years ago. Since then, advances in DNA techniques have made it possible to sequence hundreds and thousands of ancient genomes. Today, 13 years later, scientists have obtained genome data from more than 10,000 ancient humans, and data accumulation continues at an exponential rate. The vast majority of the studied ancient genomes were obtained from various places in the territory of Eurasia, which is distinguished by the huge diversity of its genes, cultures, and languages. Here we give an overview of the migration, mixing and continuity of the human population across the territory of Eurasia, starting from the period of its settlement by modern people and ending with the most mobile period in the history of mankind - the Iron Age.
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Antisocial behavior (ASB) is a complex phenotype caused by the interaction between genetic and environmental factors. In past decades several genome-wide association studies (GWAS) and their meta-analyses identified up to 500 SNPs linked to externalizing pathology in Western Europeans. However, a question on their relevance to ASB in Eastern Europeans (i.e., Russians) remains open. Therefore, the present study aimed to replicate the effect of SNPs obtained from externalizing behavior GWAS meta-analysis on homicide behavior considering a possible modulating effect of social/lifestyle factors. We have selected top six SNPs (p < 10–21) from recent GWAS meta-analysis of ASB (Karlsson Linnér et al., 2021) including CADM2 rs993137, ZIC4 rs2279829, REV3L rs458806, XKR6 rs4240671, SORCS3 rs11596214, and BDNF rs6265. Subsequent genotyping was performed in the sample of homicide offenders (N = 227, 7% women) and corresponding control group (N = 254). A series of logistic regression (PLINK v.1.09) confirmed the association of REV3L rs458806 in the total sample (p = 0.044, OR = 1.346), while SORCS3 rs11596214, ZIC4 rs2279829, XKR6 rs4240671 demonstrated their association with criminal behavior in the subgroups including smoking, low-educated offenders, individuals with psychopathologies and conflicts in families. Our findings replicated the effect of REV3L, SORCS3, ZIC4, and XKR6 genetic variants on ASB in the Russian cohort under a moderating impact of social/lifestyle factors. However, the effect of social/lifestyle factors including sex, somatic diseases, and smoking on escalating antisocial behavior exceeded that of examined genetic variants.
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The focus of this review is on evaluating the contribution of various regulatory mechanisms of calcium metabolism to the execution of key microglial functions such as patrolling, migration, proliferation, polarization, as well as mitochondrial plasticity and inflammasome assembly. We address current issues on the regulation of calcium homeostasis in microglial cells and microglia-like cells (MLCs). A concise historical overview of microglia and MLCs is provided, followed by an analysis of their functioning in both normal and pathological conditions. We refer to the functional classification of various calcium channels and transporters expressed in the plasma membrane and endoplasmic reticulum of microglia along with elucidation of the mechanisms leading to elevated cytosolic Ca2+ concentrations in microglial cell upon their activation. Then, we discuss the contribution of NAD+-glycohydrolase/CD38 to the regulation of calcium homeostasis in microglia. The review highlights contemporary approaches for manipulating microglial calcium metabolism with potential implications for the treatment of neurodegenerative diseases and neuroinflammation. Additionally, we briefly mention on modern imaging methods for studying calcium signaling in microglia. Thus, we summarize current data that shed the light on the intricate interplay between calcium regulation and microglial function in brain (patho)physiology. It also offers insights into potential therapeutic strategies and visualization techniques in the context of diagnostics and treatment of neurodegenerative disorders and neuroinflammation.
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The ability to track some structural changes in enhanced green fluorescent protein (EGFP) by observing its fluorescence makes EGFP a convenient object for studying the protein denaturation process and the influence of some factors on denaturation, in particular, the presence of nanoparticles. In this work, we studied the EGFP fluorescence during its de- and renaturation processes, as well as the influence of the addition of iron oxide nanoparticles on EGFP fluorescence and these processes. Kinetic measurements of denaturation revealed some details of this process. During renaturation, we managed to achieve a 60% recovery of EGFP fluorescence compared to the native protein. We also demonstrated significant effects of the presence of iron oxide nanoparticles. Iron nanoparticles approximately doubled the denaturation rate and suppressed protein renaturation.
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1,2,4-trioxolanes were obtained by the ozonation of fish oil with a mixture of oxygen and ozone to study their physicochemical properties. The biological activity of 1,2,4-trioxolanes with betulin from birch bark extract in fish oil was evaluated under hypoxia and immobilization stress in rats. 1,2,4-trioxolanes composition led to LPO indexes normalization (malondialdehyde, Schiff bases, diene and triene conjugates), the activation of NADP/H and NAD/H-dependent enzymes (GR, G6PDH, LDH, AlDH), as well as SOD and catalase, under stress in rats. Thus, we estimated 1,2,4-trioxolanes with betulin in fish oil to regulate oxidative and energy metabolism under hypoxia and immobilization stress in rats. The findings show that studied composition can be useful to prevent and treat the diseases caused by oxidative stress.
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The Republic of Sakha (Yakutia), located in the northeastern part of Russia, is characterized by an extremely cold climate, to which the indigenous people is adapted. Over the past decades, there has been a significant increase of the incidence of type 2 diabetes mellitus (T2DM) among the indigenous population. It is known that polymorphisms of the mitochondrial genome, in particular, the 16189C variant of hypervariable segment I (HVS-I), may contribute to the development of T2DM. The aim of the study was to assess the association of mitochondrial DNA (mtDNA) HVS-I polymorphisms with the type 2 diabetes mellitus in the Sakha (Yakut) population. Sequencing of HVS-I mtDNA in 102 patients with T2DM and 101 non-diabetic controls revealed 67 haplotypes and 64 SNP variants. There was no statistically significant difference in the frequencies of detected HVS-I polymorphisms and haplotypes between the two groups, which indicates the absence of a close association between HVS-I polymorphisms and T2DM in the Sakha population.
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Overtraining syndrome (OTS) is a condition associated with prolonged dysfunctional adaptation to physical exercise and a long-term imbalance between training and recovery that results in decreased sports performance lasting from several weeks to months with serious consequences for the health of athletes. The problem of diagnosis and prevention of OTS remains relevant, as the diagnosis is often made retrospectively. Currently, no clear reliable biochemical or functional markers for early detection of OTS are described, and the features of pathogenesis of this syndrome remain unclear. The present overview describes the basic theories of OTS development, the main biomarkers and their diagnostic significance, as well as some novel parameters and methods that may be possibly perspective for early detection of OTS.
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Issue 2 | June 2023
Given the complexity and huge variety of human diseases and areas of medicine aimed at reducing or eliminating the negative consequences of various disorders in the normal functioning of complex systems, it is important to study these complex processes in model organisms. This article provides a short overview of human diseases and some applied areas of medicine in which some progress has been made through the study of model animals. In the future, new knowledge obtained on various animal models can be used to elucidate the etiology of disorders, with subsequent implementation in clinical medicine.
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COVID-19 is a disease first reported in 2019 that claimed the lives of more than 6.5 million people worldwide, paralyzed transport links and locked the borders of many states for a long time. In 2023, 3 years have passed since, yet not all countries have fully recovered and lifted the restrictions, which, of course, highlights that COVID-19 has had a huge impact on all aspects of modern life. The pandemic has given a strong impetus to the development of science and the study of COVID-19 and infectious diseases in general around the world, many articles on COVID-19 have been published in the past 3 years. Particularly interesting was the fact that while some people were asymptomatic, had mild COVID-19, other patients required mechanical ventilation and even medically induced coma. In this regard, the study of the genetic factors contributing to the severe course of the disease, comorbidities and the individual response to drugs has become especially relevant. In our work, we consider the main genes and entire loci of chromosomes involved in the pathogenesis of COVID-19. Genes such as IFNAR2, TMPRSS2, ACE2, TYK2, DPP9, HLA, OAS3, ABO, 3p21.31 locus and 12q24.13 locus have been considered; in addition, the association of severe COVID-19 with diseases such as type 2 diabetes, cardiovascular disease and obesity was discussed.
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Bacillus cereus is an opportunistic Gram-positive anaerobic spore-forming bacterium found in various environmental conditions. One of the pathogenicity factors is hemolytic toxin II, which belongs to the group of β-barrel pore-forming toxins. HlyII B. cereus differs from other β- barrel pore-forming toxins by having a C-terminal extension of 94 amino acid residues. In this work, the binding sites of monoclonal antibodies obtained against recombinant HlyIICTD on the protein surface were determined. Using monoclonal antibodies, it was shown that HlyIICTD interacts with the erythrocyte membrane in a strictly defined manner. The localization of epitopes recognized by monoclonal antibodies suggested that the binding site of HlyIICTD with the membrane is localized in the C-terminal region of HlyIICTD molecule.
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Biomedical research often requires the use of precise equipment for micromanipulation, particularly in cellular biology. However, relatively inexpensive devices for cellular manipulation with electrophysiological methods or local chemical application with micrometer accuracy are rarely available in the equipment market. In this study, we present a method to develop a micromanipulator device based on stepper motors that is controlled by a microcontroller via a gamepad. This micro-motion system can be easily produced in any laboratory for various scientific experiments that require the movement of the electrode or pipette with a precision of several micrometers.
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Objectives: to analyze early postoperative arrhythmias in patients after surgical treatment of atrial fibrillation, correction of valvular pathology and sympathetic denervation of the pulmonary arteries in order to correct high pulmonary hypertension. Materials and methods: the study included patients with diagnosed mitral valve disease complicated by atrial fibrillation and high-grade pulmonary hypertension (40 mm Hg or more). All studied patients underwent mitral valve replacement, Maze IV biatrial surgery. In contrast to the patients in the study group, pulmonary artery denervation was not performed in the control group. On the first day after transfer to the ward from the intensive care unit, patients were fitted with a wearable cardiorecorder, which provides continuous ECG recording of one lead with automatically machine analysis with the formation of a pre-medical conclusion. Results: In all studies, a mechanism was implemented for remote ECG transmission of data in the "real time" mode for evaluation by a cardiologist. The study group demonstrated the best recovery and preservation of sinus rhythm in the early postoperative period. 92% of the patient had sinus rhythm, while in the control group this indicator was - 81% (p = 0.05). Radiofrequency denervation of the pulmonary arteries is a safe and effective method for correcting high pulmonary hypertension and helps to maintain sinus rhythm after the concomitant Maze IV procedure (p = 0.05). The use of automatic cardiac screening has demonstrated the feasibility of detecting early arrhythmias in automatic mode with the possibility of timely personalized correction of antiarrhythmic therapy and treatment tactics in general.
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Various studies have led to our present knowledge of α1-adrenoreceptors and their role in the regulation of cardiovascular physiology. Our study aimed to study the effect of the α1-adrenergic receptor agonist methoxamine on the frequency of spontaneous activity and electrical activity parameters of the atrial myocardium with a preserved sinus node and an imposed rhythm in newborn rats. In working cardiomyocytes of newborn rats with a preserved sinus node, methoxamine caused an increase in the frequency of spontaneous activity. α1-adrenergic receptor agonist methoxamine increased the duration of the repolarization phase of the action potential in both the imposed and the own rhythm.
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A comparative analysis of the effects of serotonin (200 μg/kg), dopamine (60 μg/kg), promethazine (2 mg/kg) and sulpiride (1 mg/kg) administered once intraperitoneally on heart rate variability in male non-linear rats was carried out. Serotonin and dopamine do not change heart rate, but increase the centralization index by 122% (p < 0.05), serotonin due to an increase in the power of LF waves by 136% (p < 0.05), dopamine due to a downward trend power of HF waves by 51% (p < 0.1) compared to the control. Promethazine reduces the power of LF and VLF waves by 64-66% (p < 0.05) and the index of centralization (p < 0.01), while the proportion of HF waves in the spectrum increases to 75%, which is higher than the control (p < 0.01). Sulpiride increases heart rate frequency by 25% (p < 0.01) and the index of centralization (p < 0.001), reduces the power of HF-waves by 70 % (p < 0.001) from the initial state, increases the power of LF-of waves by 336% and VLF-waves by 197% (p < 0.001), so the total proportion of LF and VLF waves increases to 85%, which is higher than the control (p < 0.05). The results suggest that peripheral serotoninergic and dopaminergic mechanisms have opposite effects on heart rate variability.
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This research evaluates particularities of surface structural organization of surgical samples of intracapsular distracted formations in treating post-traumatic fibrous ankylosis of proximal interphalangeal joints by means of scanning electron microscopy. The research is carried out on small-fragment samples of intracapsular formations collected in the course of treatment from 8 patients of the Microsurgical Department. Morphological evaluation of the surface of two types of samples was performed: that of proximal interphalangeal joint capsules, and of the regenerate itself. Structural analysis of the surgical samples allowed to identify considerable differences both in form and size of single structure-forming elements, and in their organizational types. It was established that the regenerate has a more compact structure. In the course of microanalysis of elemental composition, it was ascertained that mass percentage of calcium and phosphorus in the examined regenerate samples is 4.4 and 2.2 times respectively higher compared to the capsule samples. These results could be used in further research into approaches to the evaluation of connective tissue regeneration after surgeries, as well as in making recommendations for operative or physiotherapeutic patient treatment in cases of joint capsule distraction.
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Bronchial asthma (BA) can be accompanied by cognitive impairment (CI). However, the relationship between cognitive dysfunction and asthma has not been fully elucidated. Cognitive disorders in patients with asthma largely depend on age, asthma control impairment, the severity of symptoms of the disease, and the duration of its course. The severity of cognitive dysfunction begins to appear already in childhood and increases significantly in adult and elderly patients with asthma. Cognitive impairments can affect adherence to therapy in patients with asthma and negatively affect the achievement of BA control. There is evidence that chronic and acute hypoxia with insufficient control of BA can contribute to the development of cognitive dysfunction. Comorbid conditions also contribute to the enhancement of cognitive dysfunction in patients with asthma. It is known, for example, that asthma is associated with increased levels of depression and anxiety, which may contribute to the development of cognitive dysfunction. Increased body weight may also be a risk factor for cognitive impairment in BA patients. Timely detection of CI and appropriate correction will positively affect the results of therapy. This review examines the features of cognitive dysfunction in patients with BA of different ages, the relationship of CI to the control and duration of the disease, and the impact on the cognitive status of comorbid conditions. This evidence will allow us to streamline our knowledge about the state of cognitive function in patients with asthma for their timely diagnosis, correction, and, ultimately, improvement of BA control.
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The aim of the study was the IR spectroscopic and chromatographic analysis of products formed during the ozonation of fish oil. Fish oil samples were ozonized using an ozone therapy apparatus with an ozone destructor "Medozons - O3" (Nizhny Novgorod, Russia). The study of the chemical composition of fish oil ozonolysis products was performed on a Shimadzu IR Prestige 21 (Japan) infrared Fourier spectrophotometer in the region of wave numbers 4000 - 400 cm–1 in the form of liquid films in KBr, NaCl, or ZnSe windows. Chromatographic study of the composition of fatty acids was carried out on an Agilent 7890B gas chromatograph with a mass-selective detector 5977A. Qualitative and quantitative determination of propionic acid was carried out on the gas chromatographic complex "Chromosome GC – 1000". Chromatographic data processing was performed on the hardware and software complex "Chromatek-Analyst". The dynamics of IR spectra and quantitative composition of fatty acids (chromatographically) of the studied samples before and after ozonation were evaluated. The concentration of reactive oxygen species was controlled by iodometric titration according to the interstate standard GOST ISO 3900 – 2013. The acid number and saponification number of reaction products were determined by chemical methods. It was found that during the processing of fish oil, the reaction with ozone mainly proceeds locally along the ω–3 double bonds, leading to the formation of hydroperoxyesters and hydroperxy acids, as well as propionic acid. It is shown that the number of fragments of ω-6, ω-7 and ω-9 fatty acids practically does not change.
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The article discusses the organism`s adaptive capabilities of different age groups in the population with the use of cardiointervalography and hemodynamic parameters. There was a decrease in the total power of the heart rate variability spectrum and the power of all spectral parameters with an increasing age. In age groups 1 and 2, LF, % and VLF, % were more pronounced, there was a significant predominance of VLF, % in group 2, which reflected an increase in the level of the sympathetic link regulation activity. A more significant increase in the power of very low frequency oscillations indicates an increase in the influence of the central circuit of regulation along with an increase in the cerebral ergotropic influences. Assessment of oxidative metabolism by fluorescence spectroscopy showed an increase in the indicator in group 2, with a tendency to decrease in group 3 as compared to the values of group 1. The performed analysis of the frequency-amplitude spectrum revealed changes in the reactivity of the structures of the microvasculature depending on age. An increase in the volume of blood entering the microcirculation system was recorded, but the regulatory effect on microcirculation was different: group 2 was dominated by central mechanisms of regulation, in group 3 there was a predominance of local regulatory mechanisms. Thus, with age, there is a decrease in the adaptive reserves of the body with a tension in the regulatory vegetative homeostasis and a transition from nervous mechanisms of regulation to a lower level of humoral mechanism.
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A study was made of the general condition, changes in the relative mass of internal organs and blood parameters of C57Bl/6 mice 3 months after local irradiation of the chest. Irradiation was carried out in five equal fractions with total doses of 5, 10 and 20 Gy. Single cases of death of irradiated animals were noted in each group 2 months after irradiation. 90 days after irradiation, an increase in the relative mass of the lung was noted in all groups of irradiated animals, as well as a decrease in the relative mass of the spleen and liver. A decrease in the number of platelets in all groups after irradiation was established, as well as a change in erythrocyte indices, which was most pronounced in groups irradiated at doses of 10 and 20 Gy. An increase in the activity of transaminase enzymes in the blood serum of animals depending on the dose of irradiation, an increase in the activity of LDH and phosphatases, as well as a decrease in the level of total protein were noted, which indicates damage to the organs of the thoracic region and liver and persistent metabolic disorders in animals 90days after local irradiation.
Issue 1 | March 2023
Introduction: the average thickness of the tympanic membrane (TM) normally fluctuates around 100 µm. Most of the measurements of the TM thickness presented in previous works were performed on cadaveric material. The anatomical parameters of the TM can be measured using optical coherence tomography (OCT). Materials and methods: we used spectral-domain OCT with a non-contact probe to study the structures of the middle ear. The results of OCT examination of 58 tympanic membranes of patients without middle ear pathology were analyzed. Quantitative analysis of OCT images was performed using the ImageJ program. The thickness of the tympanic membrane was calculated manually at 5 randomly selected points. To convert optical values into metric ones, we used an average refractive index of 1.45. Results: the TM thickness in patients without ear pathology was 138 µm (±29). Conclusions: OCT is an effective method of non-invasive examination of the TM and can be successfully used to measure the TM thickness. Improving the algorithms for processing OCT images in order to automatically calculate the TM thickness and make TM topographic maps remains a crucial task and its solution is expected to significantly improve the diagnostic properties of the method.
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For a long time, the strains Bifidobacterium bifidum 1, Bifidobacterium bifidum 791, and Bifidobacterium longum 379 have been successfully used as probiotic producers; the conclusions about the possibility of their use, their role and functions in the human body were made empirically based on indirect signs. At present, high-tech equipment and modern research methods allow to conduct a complete study of the properties and characterization of strains; it is regulated by modern regulatory documents. In our work we applied the classical bacteriological method, MALDI TOF mass spectrometry method, NGS whole genome sequencing. The analysis of 27 clones of the strains revealed that they have typical morphological properties and stable biochemical profiles; the lists of ionized protein masses (so-called "strain markers") were determined. The whole genomes of strains deposited by us in GenBank in 2015-2017 were analyzed for the first time; we found that they do not contain pathogenicity genes, integrated plasmids, determinants of transmissible antibiotic resistance, and belong to phylogenetic clusters formed by probiotic-producing strains. It was shown that B. bifidum 1 and B. bifidum 791 have a pronounced ability to consume glycans of intestinal mucus, while B. longum 379 can efficiently consume plant glycans. The genomes of B. bifidum 791 and B. longum 379 strains contain genes for the synthesis of the most important neurometabolites of tryptophan and folic acid; the genome of B. bifidum 791 strain contains genes for the synthesis of lasso peptide and flavucin, class I lantibiotics with a wide spectrum of antimicrobial and antiviral activity.
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A review using the information on the problem in domestic and foreign sources of literature was carried out. Selected wild berries of Yakutia in their composition contain anthocyanins, flavanols and other biologically active compounds with experimentally proven anticancer effect in vitro against breast cancer cells. It has been noted that there is an insufficient study of wild berries as preventive agents against breast cancer. Experimental studies have proven the role of lingonberry in inhibiting the development of carcinogen-induced epithelial cancer, and cell culture of lingonberry and cloudberry can reduce the viability and proliferative activity of breast cancer cells. It is recommended to conduct researches on the effect of extracts, juice of northern berries on the development of breast cancer cell culture and find out their possible preventive and therapeutic effect.
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Cataract remains the leading cause of reversible blindness worldwide. Inflammatory postoperative complications remain a significant challenge associated with cataract phacoemulsification. These include corneal melting, also known as aseptic keratomalacia, a sight-threatening inflammatory condition that precedes corneal perforation. Case description: a 70-year-old patient underwent cataract phacoemulsification and subsequently developed indomethacin induced corneal erosion. Despite 2 months of conservative treatment, the erosion progressed to a corneal ulcer. To prevent corneal perforation, a prophylactic conjunctival flap was performed. Dry eye disease and Sjogren's syndrome were diagnosed postoperatively. Follow-up was 5 years. Conclusions: the presented case demonstrates for the first time the role of indomethacin as a trigger of corneal melting after cataract phacoemulsification. Corneal melting, potentially leading to corneal perforation, may occur after successful surgery not only in patients with diagnosed dry eye disease and systemic collagenosis but also in asymptomatic patients. Torpid progression, resistance to conventional therapy and a high risk of corneal perforation require a specialist to select appropriate therapeutic and surgical treatment methods as soon as possible. Treatment begins with withdrawal of NSAIDs, local and systemic steroid therapy, intensive corneal lubrication and objective monitoring using anterior segment optical coherence tomography.
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No doubt that the identification of predisposition toward certain types of activities becomes essential, which will allow using additional educational technologies as early as possible, thereby contributing to the development of existing powers in individuals and, as a result, increasing the productivity of future endeavors. There has been conducted a molecular genetic study of the frequency distribution of alleles and genotypes of 15 polymorphic genes variants, those involved in the regulation of memory span, synaptic plasticity, and the functioning of neurotransmitter systems of the brain of 7th and 8th grade students. DNA samples recovered from peripheral blood lymphocytes of 197 adolescents (89 boys, 108 girls), served as the material for molecular genetic testing; the students of the Municipal Autonomous Educational Institution "Lyceum No. 42" of the city district of Ufa city of the Republic of Bashkortostan - none registered in medical records. Statistically significant associations were defined: on the "general activity" scale with polymorphic loci rs6265 of the BDNF gene, rs4680 in the COMT gene and rs1387923 of the NTRK2 gene, on the "mathematical intelligence" scale - rs6265 of the BDNF gene, on the psychoticism scale - rs2710102 in the CNTNAP2 gene, rs6280 in the DRD3 gene, rs1018381 in the DTNBP1 gene and rs1387923 in gene NTRK2, on the neuroticism scale - rs6675281 in the DISC1 gene, on the “spatial intelligence” scale - rs6675281 in the DISC1 gene and rs4971684 in the NRXN1 gene, on the “extroversion” scale - rs2832407 in the GRIK1 gene.
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In-depth studies of the etiology and consequences of brain damage induced by chronic prenatal hypoxia are now urgently needed to improve the therapeutic strategies and reinforcing the adaptive capacity of nerve cells to oxygen deficiency. Here, we analyzed the characteristics of spontaneous calcium activity of primary hippocampal cultures obtained from mouse embryos exposed to chronic prenatal hypoxia in a late period of development in vitro (DIV 15 - DIV 21) and the adaptive capabilities of neuron-glial networks to the acute hypoxic injury. Chronic hypoxic stress caused several delays in the functional development of primary hippocampal cultures. On DIV 15, the cultures exhibited the spontaneous calcium activity with a decreased number of active cells and duration of Ca2+ oscillations. The reduced values of network characteristics (DIV 15) in the presence of partial destruction of the functional architecture of neuron-glial networks with a reduced force of correlated connections between cells during entire observation period were also shown. Chronic hypoxic stress altered the functional culture's response to acute oxygen deficiency. On day 7 after acute hypoxia modelling, against the background of significant decrease in the number of functionally active cells, the frequency and duration of Ca2+ oscillations did not differ from the intact values. Nevertheless, significant decrease in the network characteristics accompanied by substantial rearrangements in the functional architecture of neuron-glial networks indicate the death of significant part of the network elements and increase the risk of aggravation the synaptic transmission impairments leading to loss of network functionality in more distant post-hypoxic period.
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The paper presents the results of a study of patterns of exploratory behavior and processes of lipid peroxidation in liver tissue in young and old female rats, depending on the stage of the estrous cycle. The studies were carried out on females in the estrus and diestrus stages. It was shown that females at the age of 2-3 months were characterized by regular reproductive cycles with an average duration of 4.5 ± 0.11 days, which corresponds to the normal estrous cycle of sexually mature rats, which includes 4 consecutive stages. Animals 24 months old had cycles different from the normal physiological cycles of young animals. The work shows that in young sexually mature females the motor activity in the open field test is higher, and the level of anxiety is lower in the estrus stage compared to the analogous indicators in the diestrus stage. In young females, the baseline MDA level was significantly higher in the diestrus stage relative to that in the estrus stage. Patterns of exploratory behavior and the level of LPO depended on the age of the animal. Old rats had an increased level of anxiety regardless of the stage of the reproductive cycle, while young females were characterized by higher patterns of exploratory activity. In addition, old female rats had a higher level of free radical oxidation in comparison with young animals.
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We have analyzed the distribution of alleles and genotypes of three polymorphic variants: rs11385942 of the LZTFL1 gene, rs657152 of the AB0 gene, and rs2109069 of the DPP9 gene previously associated with COVID-19 in a sample of 80 presumably healthy individuals of the Permsky Bashkirs from the Permsky region of Russia, 48 representatives of the Burzyan region and 47 representatives of the Arkhangelsky district of the Republic of Bashkortostan of Russia. No statistically significant differences were shown between the Burzyan Bashkirs and the populations of Southern Siberia in the rs11385942 LZTFL1 locus, in contrast to the subpopulations of the Permsky and Arkhangelsky Bashkirs. We also revealed statistically significant differences in the frequency of the minor allele, the rs11385942 LZTFL1 and rs657152 loci of the ABO gene between Burzyan Bashkirs and a mixed sample of Komi and Udmurts. In turn, the subpopulations of the Permsky and Arkhangelsky Bashkirs in the rs657152 locus of the ABO gene were statistically significantly different from the Tatars.
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Issue 4 | December 2022
Optical coherence tomography (OCT) is one of the most promising, innovative and rapidly emerging intraoperative imaging modalities for neurosurgical guidance by brain tissue imaging, "optical biopsy", brain cerebral vascular detection, nerve fibers and white matter tracts detection. In this article, we provide a short survey of cross-polarization OCT and different types of OCT probes that can be used in routine neurosurgical practice. Through different types of probes there are multiple applications where OCT can play a highly complementary role in offering the real-time microscopic assessment and imaging of normal and pathological brain tissues. The biopsy-needle based probe for CP OCT was shown as an effective instrument for brain tissue mapping and express estimation of tissue status as well as for detecting large blood vessels to prevent causing bleeding during biopsy sampling. The folded CP OCT probe for intraoperative use for brain tissue examination was shown as a potentially efficient sensor head for CP OCT. The probe demonstrated high lateral resolution in diffractive limited probing beam quality. The length of the dismountable probe tip allows using the probe under operating microscope. The designed family of specialized probes allows CP OCT to occupy a niche of devices for express brain tissue examination in situ after finishing of the approvement for clinical use process.
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The effect of therapeutic technologies used in the treatment of burns on the parameters of nitrogen metabolism has not been sufficiently studied. In this regard, the assessment of the level of nitrites and nitrates of blood and tissues in combined thermal trauma (CTT) under the influence of NO is of scientific and practical interest. The aim of the study was to determine the concentration of nitric oxide metabolites in the blood and organs under the influence of NO during CTT. The experiment was carried out on white male rats of the Wistar line. CTT (contact burn on the area of 20% of the body surface and thermal inhalation exposure for 20-30 seconds) was applied under anesthesia. Animals with CTT were treated daily with intraperitoneal injections of a 10% dinitrosyl iron complexes (DNIC) solution or were inhaled with NO (20ppm). The concentration of nitrites and nitrates was determined in the blood plasma and homogenate of the liver, heart, kidneys, and lungs. CTT was accompanied by hyperproduction of NO in blood plasma on the 1st, 7th and 10th days after injury with a maximum growth on the 1st day. The largest decrease in NO2- and NO3- levels during CTT was observed in the kidneys > liver > lungs > heart. The use of DNIC in CTT led to an increase in NO2- and NO3- in the organs. Inhalations of NO during CTT increased the concentration of NO2- and NO3- in the homogenate of the lungs > kidneys > heart > liver.
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Selenium and its compounds are promising immunomodulatory agents. We studied the ability of selenium nanoparticles (Se NPs) and their combination with immunoglobulins (IgG) to influence the functional responses of immune cells and the expression profile of “stress associated”. The cytotoxic effect of Se NPs was also studied in primary and immortalized cell cultures. Se NPs were obtained by laser ablation in water followed by fragmentation. Fragmentation control was performed using acoustic and optical methods of size estimation. The size distribution of Se NPs was narrow and an average size was 100 nm. The Se NPs did not exhibit cytotoxicity against fibroblasts, hepatocytes and cell line L-929. Weak cytotoxicity was found for the HL-60 granulocyte-like culture. On a surviving culture of mouse granulocytes, no cytotoxic effect was found. The addition of Se NPs in combination with IgG can modulate the maximum and total production of ROS by murine granulocytes induced by W-peptide and PMA and modulate the proportion of granulocytes with calcium responses to Wp. Se NPs modulate the action of IgG on the physiological responses of granulocytes. In the study of gene expression, similar patterns were found. Se NPs increase the expression of HSP90, NFkb, Xrcc4, SOD2 genes; IgG enhances the effect of Se NPs, while IgG decreased the expression of these genes. This phenomenon can be explained by the interaction between Se NPs and IgG. Data of spectral methods showed the binding of IgG to Se NP surface and a partial change in the spatial structure of IgG.
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Physico-chemical characteristics of optical-induced break-down depend weakly on the protein concentration. This factor distinguishes lysozyme from other aqueous solutions which contain objects with nano-size. The laser radiation of the lysozyme aqueous solution during different time periods shows patterns that are quite similar to the classical patterns. In this particular paper, it has been observed that the catalytic activity of lysozyme is decreasing after the laser radiation. The optical density of the protein solutions is increasing. The fluorescence intensity of the lysozyme solution is determined by the amino-acid residues. It decreases before and after the laser light influence. The position of the maximum of the excitation and emission did not change. The shape of the fluorescence signal on the 3D maps also does not change significantly. This observation suggests the degradation of the amino-acid residues. The viscosity and the pseudo plasticity of the lysozyme aqueous solutions increased. However, there is no massive damage of the polypeptide chain. On the contrary, the optical break-down shows the intensive forming of the lysozyme aggregates.
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Control of the processes of survival and differentiation of immature neurons - non-newly generated immature neurons - nng-Ins - in the cortical areas of the brain is important for preventing the de-velopment of neurological dysfunction in disorders of brain development and physiological aging. We used olfactory stimuli (OS) in the dynamics of postnatal development (P21, P60) in CD1 mice. They have been exposed to the protocol of simulation and assessment of the piriform cortex activation upon olfactory stimuli presentation. Then, at 2 hours, 24 hours, and 7 days after stimuli presentation, we analyzed parameters of learning and memorization, social recognition, anxiety as well as the patterns of expression of nng-INs markers (DCX, PSA-NCAM), proliferation marker (Ki67), marker of postmitotic cortical neurons (Tbr1), and immediate early gene c-fos expression as a marker of neuronal activation. We found that in the period from 2 to 60 days of mice postnatal development, proliferating and non-proliferating cells co-expressing DCX and PSA-NCAM were present in the piriform cortex (PC) and responded to the presentation of olfactory stimuli. Activation of nng-INs (DCX+ PSA-NCAM+ Ki67-) in the OS-stimulated brain plasticity is more evident in the immature developing PC, whereas appearance of mitotically active neuroblasts (DCX+ PSA-NCAM+ Ki67+), presumably, coming from other neurogenic niches of the brain upon OS-driven PC activation as well as stimulus-induced differentiation of locally present nng-INs might dominate in the mature piriform cortex (P60). Thus, immature neurons in the PC might contribute to the brain plasticity in the early ontogenesis.
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The high risk of ovarian cancer is primarily associated with mutations in BRCA1 and BRCA2 genes. However, mutations in these explain only a small proportion of cases. Mutations in other genes are also involved in the disease. As a result of previous exome sequencing of DNA samples from breast cancer Germany patients with clinical signs of a hereditary form of the disease without major mutations in the BRCA1, BRCA2, CHEK2 and NBN genes, potentially pathogenic genetic variants in new breast and ovarian cancer candidate genes were selected. Selected as a result of bioinformatics analysis genes are involved in vital cell signaling pathways such as repair, apoptosis, cell cycle regulation, cell proliferation, migration, differentiation, as well as immune response and inflammation. Recently, biological microarray technologies have been widely used to study the general genetic variability throughout the human genome in order to determine genetic associations with the disease and search for genes involved in the pathogenesis of multifactorial pathologies. The use of such approaches can be very useful for identifying risk markers for the development and severity of diseases. Our case-control study is aimed at researching potentially pathogenic variants selected as a result of exome sequencing of DNA samples from Caucasian patients using microarray technology Fluidigm to assess their contribution to ovarian cancer pathogenesis in Bashkortostan. Most of the researched alleles were found with different frequencies among cases and controls; however, our data indicate that the researched potentially pathogenic variants do not contribute to ovarian cancer pathogenesis in Bashkortostan populations.
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Integration of Immature Neurons Maintain Unidirectional Connectivity of Pre-Existed Network In Vitro
Recovery of the neural networks after brain injury can be studied in vitro using cell patterning technologies such as microfluidics. In this study, neural tissue recovery of cortical networks was modeled in a three-chamber microfluidic chip by growing two weakly coupled neuronal networks and by plating new immature cells between it. The direction of synaptic connections was formed by the asymmetric design of the microchannels in the chip. We investigated dynamics of morphological characteristics of neurites growth through the microchannels and spiking activity propagation between the networks after integration of new cortical cells. The results can be used for the development of new approaches for brain functional recovery after injury.
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The article considers the development of biocomposite materials as bacterial cellulose-based hydrogel, chitosan and alginate, plus physiologically active compounds – fusidic acid, resveratrol and dihydroquercetin. It has been found that the use of hydrogel systems derived from microbial polysaccharides and containing bacterial cellulose (BC) / sodium alginate with CaCl2 /sodium fusidate (SF) and BC/chitosan /sodium fusidate helps to reduce the intensity of lipid peroxidation processes and stabilize phospholipid and fatty acid composition of the skin. It is consistent with the findings of the authors on longer release of sodium fusidate from biocomposite materials of this composition. Therefore, the use of BC and chitosan in combination with sodium fusidate, which exhibits antibacterial properties, and the crosslinking of sodium alginate with calcium chloride solution proves to be the most effective for restoring the skin’s lipid composition and shortening the course of treatment. Most likely, this effect must be explained by the constant release of physiologically active compounds from hydrogel composites and its impact on damaged skin areas.
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In the last decade, glial cells, primarily astrocytes, have received increasing attention in studying various aspects of brain functioning. Although astrocytes are not electrically excitable cells, they are able to generate and transmit Ca2+ signals, which can propagate from cell to cell, forming «calcium waves». Calcium signaling allows astrocytes to interact with each other, as well as regulate neuronal function. Despite numerous studies on calcium events in astrocytes under various pathological conditions, the features of astrocytic calcium dynamics in aging are currently understudied. Our work aimed to analyze the features of calcium activity in primary astrocyte cultures during physiological and accelerated aging, as well as to assess the effect of hypoxic damage on calcium activity. For this purpose, we developed an algorithm for the accurate detection of calcium events and the calculation of indicators describing the main parameters of calcium events in a cell. It was shown that the frequency of calcium event generation in astrocytes decreases during aging while their duration and amplitude increase. Hypoxia aggravates the changes caused by cell aging.
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The visceral system can influence consciousness and emotions. In this paper, we investigated whether the processing of short-term emotional stimuli along different phases of the cardiac cycle is selectively regulated. Emotional and neutral faces were presented to the volunteers during cardiac systole, when the release of blood from the heart causes arterial baroreceptors to centrally signal the strength and time of the heartbeat, and in diastole, the period between heartbeats when the baroreceptors are at rest. Participants passively observed the faces appearing in the oddball paradigm in immersive virtual reality (VR). Combining electroencephalography and electrocardiography, cardio synchronous ERP was recorded. The ERP data at the occipital Oz electrode demonstrate the dependence of emotional stimuli in respect to the cardiac cycle. ERP in VR were higher during diastole than systole. More specifically, the cardiac phase influenced late component P300 of the ERP, did not influence early P100 and affected P200 only to angry face. This new evidence that events related to cardiac function can modulate emotional perception in virtual reality might be one more prime example of how body feedback shapes emotions.
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Erythrocytes, performing their basic oxygen transport function, simultaneously affect blood viscosity by changing the deformability of their structure, and also have a vasodilating effect on the walls of blood vessels using NO. An important role in this regulation is played by the purinergic signaling system, which was confirmed by this study of the morphofunctional parameters of red blood cells in the presence of ATP and sodium nitroprusside (SNP). We found that under the action of 0.5 mM ATP on red blood cells, the ability of hemoglobin (Hb) to bind oxygen decreases against the background of a slight increase in complexes with oxygen, while the total number of membrane-cytoskeletal proteins also decreases. This, in turn, is accompanied by a redistribution of Hb molecules and an increase in the area of red blood cells. In the presence of 5.0 mM ATP, the oxygen transport function of erythrocytes and the quantitative composition of membrane proteins change similarly, while the geometric height and volume of the cells are significantly reduced. The combined effect of 100 μM SNP and 0.5 mm ATP has the greatest effect on the conformation of hemoporphyrin molecules, which leads to a sharp increase in hemoglobin complexes with oxygen, while the affinity of hemoglobin for oxygen decreases. The membrane-cytoplasmic component of the cell also undergoes changes. Thus, ATP, both separately and in combination with SNP, affects the oxygen transport and regulatory function of red blood cells, activating the purinergic signaling pathway and triggering a cascade of adaptation reactions in the cell.
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The paper focuses on studying the rheological properties of erythrocytes in patients with a new coronavirus infection in the acute period and during early convalescence. A significant decrease in erythrocyte membrane plasticity was found in all patients in the acute period of the disease and every second recovering patients in the early post-COVID period. An increase in the degree and rate of erythrocyte aggregation in a quarter of cases, regardless of the observation period, was accompanied by inhibition of erythrocyte disaggregation mechanisms, which can serve as a pathophysiological basis for the formation of hemorheological disorders both in the micro- and macrovasculature in the first 3 months after recovery.
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This year Nobel Prize in Physiology or Medicine awarded to Swedish paleogeneticist Svante Pääbo demonstrated the significance of ancient genomes investigation, shedding bright light to the aDNA science. As an ancient DNA study has evolved from the 1980s to the present days it has experienced peak of inflated expectations, dramatic fall to trough of disillusionment and slope of enlightenment with exponential data accumulation after the successful introduction of NGS approach. Today ancient DNA study is a rapidly developing and facilitated by advanced technologies science that challenges our assumptions about the past by analyzing over a million-year-old ancient specimens. Here we provide an overview of peculiar properties of ancient DNA analysis of human and microbes genomes.
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Issue 3 | September 2022
B. pumilus metalloproteinase was firstly isolated and characterized by Kazan Federal University scientists. Primary structure analysis showed that the novel enzyme has no analogs among prokaryotic enzymes and occupies an intermediate position between two large families of the metzinkin clan metalloproteinases – adamalysins and astacins. These families are mainly represented by eukaryotic enzymes, which play an important role in human life and health. A more detailed study of the structure and functions of novel metalloproteinase requires an efficient expression system. B. pumilus metalloproteinase gene (mprBp) was cloned into the pGP382 expression vector under a strong constitutive promoter of the degQ36 gene (PdegQ36). The resulting construct was used to transform B. subtilis Δ6 strain. This strain was constructed by CRISPR/Cas9 genome editing technology with deletion of some prophage genes of B. subtilis 168. The functional role of prophage genes is poorly understood. It is possible that prophage deletion will increase the expression of secreted enzymes. For the transformed strain we determined the dynamics of growth and accumulation of proteolytic activity by hydrolysis of azocasein. The dynamics of proteolytic activity accumulation by this strain has a different character in contrast to the protease-deficient strains carrying the gene of the investigated enzyme. The result of this work was to obtain an effective producer strain of adamalizin-like metalloproteinase of B. pumilus, which can be used in the production of the enzyme for subsequent studies.
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Along with the wide spread of bacterial antibiotic resistance over the world, the treatment efficiency of infectious disease is greatly affected by the mixed biofilm formation by pathogenic bacteria. Staphylococcus aureus and Pseudomonas aeruginosa, a frequent cause of nosocomial infections, exhibit both synergistic and antagonistic interactions in co-culture, leading to various changes in the metabolic profile of bacteria, which in turn affect their sensitivity to antimicrobials. Here we show that S. aureus cell-free culture liquid exhibits bacteriostatic properties and increases the efficacy of antimicrobials against P. aeruginosa. Thus, the MICs of amikacin, gentamicin, and ciprofloxacin decreased 2-4 fold in the presence of cell-free supernatant of S. aureus 24 h culture. Furthermore, the combination of the latter with antimicrobials increased the efficacy of amikacin up to 64-fold. Thus, the combined use of cell-free culture liquid of S. aureus with broad-spectrum antibiotics can be used to increase the effectiveness of antimicrobial therapy of P. aeruginosa.
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The problem of the influence of inflammation on the processes of higher nervous activity is still relevant. In particular, it implies an analysis of the safety and effectiveness of biological regulators’ effects with neurotropic and immunomodulatory potential. The study examined the effects of Salmonella tiphy LPS on behaviour and cognitive function in adult male Wistar rats with different levels of melatoninergic system activity, using various tests. It was found that daily injection of LPS to rats of the group А (50 μg/kg, i.p.) for 10 days leads to the formation of a special pattern of behavior with a low level of tentative research activity in the presence of increased anxiety. The rats of the group В received combined exposure to LPS (50 μg/kg, i.p.) and melatonin (5 mg/kg, orally) distinguished a higher level of various variants of motor and research activity in conditions of relatively low anxiety in contrast to the rats of the group А. The paper discusses the features and mechanisms of the formation of "painful" behavior in an experimental model of inflammation, depending on the level of activation of the components of the melatoninergic system.
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Microbial production of testosterone from phytosterol is a promising alternative to the chemical synthesis from androstenedione, which is currently used. It is possible using wild-type or recombinant strains of Mycolicibacterium neoaurum. One of the key steps of biotechnology development is inoculum preparation which determines the volumetric/mass ratios of the bioreactors and affects the biotransformation rate and duration. In the present work, the conditions for the growth of testosterone-producing strain of Mycolicibacterium neoaurum recombinant strain with enhanced 17β-HSD activity were optimized. The effects of medium composition, temperature, and glucose supplements on biomass size and density were estimated. The results are of importance for the development of microbial technology for testosterone production.
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Noroviruses with the GII.P16 polymerase gene have long been considered rare. However, since 2015, there have been reports from different geographical regions about the spread of new recombinant strains of norovirus in which GII.P16 polymerase is associated with a capsid protein of various genotypes. In the autumn of 2016, a sharp increase in the frequency of detection of noroviruses was observed in Nizhny Novgorod, which coincided with the appearance of new recombinants GII.4_Sydney and GII.2 – with GII.P16 polymerase. Based on the sequences of the RNA-dependent RNA polymerase gene present in the GenBank database, a phylogenetic analysis of noroviruses with GII.P16 specificity was performed by constructing Bayesian phylogenetic trees. Analysis of amino acid sequences showed that representatives of the novel variant of the GII.P16 polymerase gene had five substitutions compared to earlier strains. These substitutions are located near sites responsible for the enzyme activity of polymerase and could affect the transmissivity of the virus. Acquisition of a novel variant of the GII.P16 polymerase gene by noroviruses with different capsid protein genotypes probably provides certain advantages for recombinants and creates prerequisites for their wide distribution.
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