[Physiology of autonomic regulation of kidney and bladder functions and its clinical significance].

The results of a search in PubMed and Elibraru.ru databases using the keywords "autonomic regulation", "kidney function", "bladder function", "ECG monitoring", "PET/CT of the brain" are presented in the review. A regulation of bladder functions, control of blood pressure, heart rate and specialized functions of the nephron are discussed, which are in close relationship with the stem and cortical centers of the brain. The review presents an update at their cause-and-effect relationship and the place of each system in the formation of the general autonomic tone. The proposed integrative approach to the study of this problem will reveal previously unknown autonomous properties of the organs that form this physiological axis and determine the role of cortical dysfunction in the development of visceral pathology, which is fundamentally important for understanding the mechanisms of formation and recurrence of many urological diseases.

[The characteristics of elderly citizen of perceiving communication effects of issues of formation of healthy life-style].

The article discusses results of communication effects of various information actors on elderly citizen stimulating healthy life-style. The purpose of the study was to establish characteristics of perception by elderly Russians communication effects targeting formation and maintenance of healthy life-style, to determine degree of significance of communication channels by which such measures can provide high degree of individual motivation. The contradiction was established. From one hand, life span increases due to achievements of science, medicine and technologies promoting decreasing diseases' mortality and improving health indicators.From the other hand, the need in larger volume of caring services at lesser number of professional personnel providing social care and medical personnel comes up. It is established that age principle is one of the basic ones in segmentation of health state. According it, population of elderly age consists of several main groups having different social characteristics concerning conditions of life, medical indications, consumer behavior and information perception. The elderly pre-retired, elderly relatives, early retired, socially active retired. It is concluded that communication impact on elderly citizens concerning formation of healthy lifestyle and promotion of health saving practices are to be implemented considering specifics of particular communication channels (both interpersonal and mass ones). It is recommended in planning and implementing socially-oriented marketing and educational campaign not only to segment target audiences of senior citizens, but to evaluate degree of significance of communication channels for each of them.

[Pilot study of the manifestations of synchrony of the anatomical-metabolic and functional state of the brain, kidneys and bladder].

im. The results of clinical observations and scientific studies of recent years indicate that abstinence from urination before the introduction of radiopharmaceuticals for PET / CT procedures is accompanied by a significantly lower flow of it into the bladder than after preliminary urination. PURPOSE OF THE STUDY: Comparative analysis of PET/CT metabolism of 18F-fluorodeoxyglucose at the level of the cingulate gyrus of the brain, different regions of the kidney parenchyma and in the bladder wall in individuals without nephrourological history, depending on the phase of the functional state of the bladder. MATERIALS AND METHODS: The results of PET/CT of the whole body of 30 patients of the radiological center of the Tyumen Regional Oncological Dispensary, including 16 men and 14 women (median age 52.5 (38; 63) years), were analyzed, distributed randomly in three equal study groups: after urination, in the intermictory period and immediately after the appearance of the first urge to urinate. RESULTS: Signs of synchrony were revealed in the anatomical and metabolic activity of the cortical vegetative centers, certain regions of the renal parenchyma and the wall of the bladder in the process of urination, the movement of urine along the urinary tract, the stage of its accumulation in the bladder and preparation for urination. CONCLUSION: The data obtained allow us to assume that the analyzed regions are equivalent areas of the general regulatory system that provides physiological processes in the organs of the urinary system. However, this fact requires further research and clarification.

Thermostable Lichenase from Clostridium thermocellum as a Host Protein in the Domain Insertion Approach.

Clostridium thermocellum lichenase (endo-ß-1,3;1,4-glucan-D-glycosyl hydrolase, EC 3.2.1.73 (P29716)) has been tested for the insertion of two model fluorescent proteins (EGFP and TagRFP) into two regions of this enzyme. Functional folding of the resulting proteins was confirmed by retention of lichenase activity and EGFP and TagRFP fluorescence. These results convincingly demonstrate that (i) the two experimentally selected lichenase loop regions may serve as the areas for domain insertion without disturbing enzyme folding in vivo; (ii) lichenase permits not only single but also tandem insertions of large protein domains. High specific activity, outstanding thermostability, and efficient in vitro refolding of thermostable lichenase make it an attractive new host protein for the insertional fusion of domains in the engineering of multifunctional proteins.

Expression of Soluble Active Interferon αA in Escherichia coli Periplasm by Fusion with Thermostable Lichenase Using the Domain Insertion Approach.

A recombinant DNA in which the interferon αA (IFN-αA) gene sequence is integrated into a loop region of the gene coding thermostable lichenase was constructed. This approach of insertion fusion with thermostable lichenase is advantageous in terms of increasing the solubility, stability, and production of the fusion partner in soluble form in general and in the periplasm of bacterial cells in particular. Thus, the insertion of IFN-αA into the loop (53 a.a.) of thermostable lichenase from Clostridium thermocellum resulted in effective expression of the soluble form of the recombinant protein in the periplasm of Escherichia coli without any compromise in biological activity of IFN-αA, while the thermostable lichenase retained its ability for functional folding without dramatic loss of its basic activity and thermostability.

Clostridium thermocellum thermostable lichenase with circular permutations and modifications in the N-terminal region retains its activity and thermostability.

The Clostridium thermocellum lichenase (endo-ß-1,3;1,4-glucan-D-glycosyl hydrolase) displays a high thermostability and specific activity and has a compact protein molecule, which makes it attractive, in particular, for protein engineering. We have utilized in silico analysis to construct circularly permuted (CP) variants and estimated the retained activity and thermostability. New open termini in the region of residues 53 or 99 in two lichenase CP variants (CN-53 and CN-99) had no effect on their activity and thermal tolerance versus another variant CP variant, CN-140 (cut in the region of residue 140), which displayed a dramatic decrease in the activity and thermostability. Construction and further activity and thermostability testing of the modified lichenase variants (M variants) and CP variants with peptides integrated via insertion fusion have demonstrated that the N-terminal regions in the lichenase catalytic domain (53 and 99 amino acid residues) that permit circular permutations with retention of activity and thermostability of the enzyme as well as the region between the C and N termini of the native lichenase in thermostable and active lichenase variants (CN-53 and CN-99) may be used for integrating small peptides without the loss of activity and thermostability. These findings not only suggest that CP predictions can be used in search for internal integration sites within protein molecule, but also form the background for further enzymatic engineering of the C. thermocellum thermostable lichenase aiming to create new fusion proteins.

[The Manifestation of the Anxiety during Fear Conditioning in Wistar Rats].

In order to identify the correlation between anxiety and conditioned fear, the behavior of the same male Wistar rats was compared in three anxiety tests (open field, light-dark box and elevated plus-maze) and in Pavlovian auditory fear conditioning paradigm using correlation, factor and variance analyses. The correlation between anxiety/bravery and locomotion indexes in different tests was not revealed. Positive correlations between grooming, urinations and defecations, rearing in three tests were revealed. These data suggest that animals reacted to various tests differently, resulting, apparently in the emergence of different anxiety levels, specific for each test. Vegetative reactions, inclination to exploration and substituting behavior were more stable characteristics of rats. Anxiety behavior in elevated plus-maze correlated to freezing response to context after fear conditioning, while high-anxiety rats had higher level of freezing to context than low-anxiety rats. The higher freezing response to sound after fear conditioning was found in rats with middle locomotor activity in open field. Conditioned fear to the context and to the sound was associated with different forms of rat anxiety during different tests.

[Influence of D1, D2 Receptor Blockade in Basolateral Amygdala on Behavior of Rats with High or Low Levels of Anxiety and Fear].

The influence of bilateral D1 or D2 receptors antagonists infusions into the rat basolateral amygdala on anxiety, as well as the expression, extinction and re-learning of conditioned fear was studied. Subjects were the male Wistar rats with high and low anxiety behavior in elevated plus maze, and also rats with low and high freezing responses during fear conditioning. The infusion of D1 receptor antagonist (SCH23390, 1 µg/0.5 µL in each side) reduced the expression of the conditioned fear to sound in rats with low freezing level, accelerated fear extinction and impaired re-learning in all animals. The injection of D2 receptor antagonist (raclopride, 1 µg/0.5 µL in each side) accelerated the extinction of conditioned fear to contextual cues in all rats and had a weak anxiolytic-like effect on behavior of high anxiety rats in elevated plus maze. These findings testify to the role of D1 receptors in the acquisition, expression and extinction of conditioned fear to stimuli, and D2 receptors in the occurrence of anxiety and fear to the contextual cues. There was discovered different sensitivity of animals with different levels of anxiety and fear to the infusion of dopamine receptors antagonists in the amygdala that suggested the inequality of dopaminergic transmission in the amygdala of animals with individual differences.

[DIFFICULTIES OF DIFFERENTIAL DIAGNOSTICS OF SEVERE COMMUNITY-ACQUIRED PNEUMONIA].

Severe community-acquired pneumonia implies the necessity of its early differential diagnostics from a wide range of nosological forms requiring not only therapeutic but also surgical treatment. The authors consider the causes of hypodiagnostics of community-acquired pneumonia and illustrate difficulties of differential diagnosis of this condition by concrete examples.

[Effects of intrabasolateral amygdala infusions of GABAA-receptor agonist and antagonist on expression and extinction of conditioned fear in rats with different freezing duration].

The rats were divided into two groups (little and much freezing animals) on the grounds of differences in freezing times after fear conditioning. Effects of bilateral intrabasolateral amygdale infusions of GABAA-receptors agonist (muscimol, 0.1 microg/0.5 microL in each side), antagonist (bicuculline, 0.07 microg/0.5 microL), saline (control, 0.5 microL) on expression and extinction of fear were compared in two rats groups. Muscimol infusion before retention test led to a decrease the time of freezing, it maximal biased on fear expression in little freezing rats. Bicuculline infusion increased aggressiveness of rats, decreased freezing time, induced elements of panic behavior, and evoked maximal behavioral changes in much freezing animals. Muscimol or bicuculline infusion before extinction training resulted in facilitation of extinction in much freezing rats, but not effected in little freezing animals. Received results testified to non-equivalence influences of GABA-receptors agonist and antagonist in animals with different fear levels and supposed differences in GABAergic receptors.

[Morphophysiological and biochemical characteristics of potato plants with various expression rates of the Δ12 acyl-lipid desaturase gene].

This paper reports on morphophysiological and biochemical characteristics of control and potato plants (Solarium tuberosum L., Skoroplodnyi cultivar) transformed with the Δ12 acyl-lipid desaturase gene (desA) grown long-term in vitro. The transformed plants showed faster growth and faster ontogenesis as compared to controls, which was accompanied with changes in the accumulation of photosynthetic pigments (chlorophylls a and b, carotenoids) and phenolic compounds, including flavonoids in the leaves. These characteristics were pronounced to a high degree in Line II plants with high expression rates of the desA gene, whereas Line I plants (moderate expression rate) were similar to control plants in many parameters.

[Range of Gene candidates involved in biosynthesis of laccase from basidiomycete Trametes hirsuta].

A comparative analysis of transcripts from the basidiomycota T. hirsuta grown with and without an inducer of the laccase biosynthesis was carried out. Methods of subtraction hybridization and massive parallel sequencing were used for this purpose. Unique transcripts encoded by genes that have a relatively high level of expression and belong to different gene ontology categories were identified. Also, a large number of transcripts were found to encode for predicted proteins, as well as noncoding transcripts. The latter may represent regulatory RNA molecules. Transcripts that increase their abundance when the laccase synthesis is induced are selected as gene-candidates involved in the laccase biosynthetic pathway.

[Effects of intra-amygdala injection of GABA(A)-receptor agonist and antagonist on respiratory and cardiac components of conditioned fear in rabbits].

The role of GABAergic system of amygdala in vegetative and behavioral manifestations of conditioned fear was studied in groups of active and passive rabbits, selected upon testing in open field and light-dark chamber. During training, the light (4 s) was paired with electrodermal stimulation of hind limbs(1 s, 10 Hz). Conditioned fear was judged by heartbeats and respiratory rates before and in the process of conditioned and unconditioned stimuli presentation. Solutions of GABA(A)-receptor agonist (muscimol, 0.1 µg/µL) or antagonist (bicuculline methbromide, 0.5 µg/µL) were applied to the right or left basal nucleus of amygdala after conditioning. Muscimol injection evoked more powerful vegetative changes in passive rabbits than in the active ones; respiratory deceleration during freezing in response to conditioned stimuli was eliminated and the reaction to electrodermal stimulation was reduced or altered. Activation of the left amygdala by bicuculline resulted in the increased probability of active locomotor reactions in active rabbits, activation of the right amygdale, on the contrary, increased probability of freezing. The results evidence the non-equivalence of amygdalar GABAergic system in active and passive animals.

[Expression of heterologous genes in plant systems: new possibilities].

The basic methods used in current practice for stable and transient expression of heterologous genes in plants are presented and compared. The key areas of research in the heterologous expression of genes in plants have been identified by analyzing literature and experimental data: modeling of metabolic pathways; creation of marker-free transgenic plants; the search for new regulatory elements and plant genes influencing the efficiency of expression of heterologous genes in plants; development of new methods for analyzing of transgenic plants and new approaches to the expression of heterologous genes in plants.

[Set of module vectors for stable or transient expression of heterologous genes in plants].

A set of module vectors for stable or transient gene expression in plants was constructed with regard to the majority of factors ensuring efficient heterologous gene expression in plants. The vectors are convenient to clone new regulatory elements and genes of interest via simple molecular cloning procedures. The vectors can be used to obtain transgenic plants with stable heterologous gene expression as well as to achieve transient expression because one vector includes the gene for the tomato bushy stunt virus p19 protein, which acts as a suppressor of posttranscriptional gene silencing.

[Effects of intra-amygdala injection of GABA(A)-receptor agonist and antagonist on behavior of active and passive rabbits in negative emotional situations].

The effects of right-side or left-side intra-amygdala injections of the GABA(A)-receptor agonist muscimol hydrobromide (0.1 microg/1 microl) and antagonist bicuculline methiodide (0.05, 0.1, 0.5 microg/1 microl) on the behavior of active and passive rabbits were studied in open field, light-dark test and during presentation of emotionally significant stimuli. The effect of compounds injection was differed in active and passive rabbits. The active rabbits were more sensitive to bicuculline injection and the passive rabbits to muscimol administration. Bicuculline induced anxiolytic-like effects on the active animals in open field, light-dark test and motor-activating effects during emotionally significant stimuli. Muscimol induced anxiolytic-like effects on the passive rabbits and sedative effects on the both groups of rabbits. The differences in effectiveness of right-side and left-side intra-amygdala injections on behavior of rabbits were revealed: more powerful changes were during injection of bicuculline in left and muscimol in right amygdala. Anxiolytic-like effects were revealed during injection of bicuculline into left amygdala of active rabbits and muscimol into right amygdala of passive rabbits. These findings indicate that there are individual-typological and interhemispheric differences in functioning of GABAergic system of amygdala.

Effects of Social Isolation on the Development of Anxiety and Depression-Like Behavior in Model Experiments in Animals.

This review describes the role of social isolation in the development of anxiety and depression-like behavior in rodents. The duration of social isolation, age from onset of social isolation, sex, species, and strain of animals, the nature of the model used, and other factors have been shown to have influences. The molecular-cellular mechanisms of development of anxiety and depression-like behavior under the influence of social isolation and the roles of the HHAS, oxidative and nitrosative stress, neuroinflammation, BDNF, neurogenesis, synaptic plasticity, as well as monoamines in these mechanisms are discussed. This review presents data on sex differences in the effects of social isolation, along with the effects of interactions with other types of stress, and the roles of an enriched environment and other factors in ameliorating the negative sequelae of social isolation.

[The Dagestan gene pool: polymorphism of immunogenetic and biochemical markers in Kumyks].

This study is a part of long-term investigations devoted to the analysis of the gene pool of Dagestan ethnic groups. The phenotype (in %), gene, and haplotype frequencies in Kumyk ethnic group are reported. A total of 39 alleles and six haplotypes of 14 loci (AB0, Rhesus, P, Levis, Kell, HP, GC, C'3, TF, 6PGD, GLO1, ESD, ACP, and PGM1) of immunobiochemical genetic marker systems were examined. Rare haplotypes of the Rhesus system were identified, including CDE in the Karabudakhkent population with the frequency of 0.030, and Cde and cdE in the Dorgeli population with the frequencies of 0.034 and 0.38, respectively. Similarly to the other ethnic populations of Dagestan examined, Kukyk populations carried rare, albeit typically "Caucasoid" gene ACP1(c) of the AcP1 locus. The frequency of this allele in the two populations was similar, constituting 0.031 for Karabudakhkent and 0.032 for Dorgeli. In Kumyks, allele frequencies of the AB0, Rhesus, P, Lewis, Kell, HP, GC, C'3, TF, 6PGD, GLO1, ESD, ACP, but not PGM1, systems were similar to the mean allele frequencies at these loci observed in the other ethnic groups from the Dagestan, Caucasus, and the whole European historical ethnographic province. At the same time, the allele frequency values obtained were different from those for the populations of Kazakhstan, Central Asia, Siberia, and the Ruswsian Far East. Thus, the results obtained for classical genetic markers indicate that Kumyks are genetically closer to the indigenous populations of Dagestan than to Turkic-speaking populations. Analysis of the fit of the observed phenotype frequencies to the Hardy-Weinberg expectations showed that compared to other indigenous populations of Dagestan examined, in Kumyks the genetic state of the population upon random allele association was close to equilibrium. Probably, this state was determined by practical absence of the consanguineous marriages upon preservation of intra-aul endogamy.

[Interaction of amygdalar neurons of active and passive rabbits in negative emotional situations].

Interaction of basal and central nuclear neurons of amygdala was studied by plotting histograms of crosscorrelation in passive and active rabbits exposed to emotionally significant stimuli. The behavior of animals was studied in the open field, light-dark test and during presentation of emotionally significant stimuli. Rabbits of different typological groups applied a certain behavioral strategy in a variety of behavioral tests. Intergroup differences were revealed in the interaction of neighboring cells of amygdala. Passive rabbits (as comparied to active rabbits) demonstrated more excitatory connections and less inhibitory connections with the latency from 50 to 150 ms. Interactions with the delta1-range and theta2-range frequencies in passive animals were more rarely observed. The asymmetry of the interhemispheric neuronal interaction in amygdala with the right dominance was revealed in passive but not active animals. The results testify that amygdala is involved in the choice of behavioral strategy, and the level of its activation is higher in passive animals.

[Influence of anxiolytic afobazole on amygdalar neuronal activity and behavior of rabbits in emotionally negative situations].

The influence of systemic injection of anxiolytic afobazole (1 mg/kg) on interaction between neurons of the basal and central amygdaloor nuclei and behavior of rabbits in negative emotional situations was studied in order to reveal characteristics of amygdalar neuronal network properties during unconditioned fear. An afobazole injection results in a prolongation of stay in the light compartment of dark-light chamber, in a decrease in freezing probability after exposure of emotional stimuli, in appearance of some aggressive reactions in open field. Behavioral alterations were accompanied by duration increase and latency changes of amygdalar neuronal inhibitory interactions, by increase of probability of neuronal interactions on delta-range frequency, by decrease of interactions on theta-range frequency. Under the influence of afobazole the asymmetry with right dominance in interhemispheric interaction of amygdalar neurons decreased. Changes in network properties ofamygdalar neurons during afobazole injection testify to reduction of activation level of this structure taking place at the expense of enhancement of inhibitory interactions between neurons.