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The effect of chronic overcrowding on the social behavior of adult male Wistar rats was studied. From postnatal day 30 (P30) to P180, the rats lived under standard (STND) or overcrowded (CRWD) conditions. Starting from P100, rat behavior was studied in the social preference and tube dominance tests, and aggressive behavior was investigated in the resident-intruder test. After decapitation of rats on P180, amygdala, dorsal hippocampus, ventromedial hypothalamus, and medial prefrontal cortex were collected and analyzed for expression of the IL-1β, TNF, TGF-β1, and IL-6 mRNAs by quantitative polymerase chain reaction. Compared to the STND group, rats from the CRWD group demonstrated shorter interaction time with a social object in the social preference test. They also had more wins in the tube test and initiated more attacks in the resident-intruder test. Expression of the IL1β gene in the hippocampus and medial prefrontal cortex and of the TGFβ1 gene in the hippocampus, amygdala, and prefrontal cortex was increased in the CRWD group. The stress induced by overcrowding increased social dominance and aggressiveness and decreased social motivation in rats. The changes in the social behavior of CRWD rats were accompanied by upregulation of expression of genes for the proinflammatory cytokine IL-1β and the anti-inflammatory cytokine TGF-β1 in a number of brain structures, which can be considered as manifestations of neuroinflammation and compensatory processes, respectively.

Background: Spreading depolarization (SD), underlying mechanism of migraine aura and potential activator of pain pathways, is known to elicit transient local silencing cortical activity. Sweeping across the cortex, the electrocorticographic depression is supposed to underlie spreading negative symptoms of migraine aura. Main information about the suppressive effect of SD on cortical oscillations was obtained in anesthetized animals while ictal recordings in conscious patients failed to detect EEG depression during migraine aura. Here, we investigate the suppressive effect of SD on spontaneous cortical activity in awake animals and examine whether the anesthesia modifies the SD effect., Methods: Spectral and spatiotemporal characteristics of spontaneous cortical activity following a single unilateral SD elicited by amygdala pinprick were analyzed in awake freely behaving rats and after induction of urethane anesthesia., Results: In wakefulness, SD transiently suppressed cortical oscillations in all frequency bands except delta. Slow delta activity did not decline its power during SD and even increased it afterwards; high-frequency gamma oscillations showed the strongest and longest depression under awake conditions. Unexpectedly, gamma power reduced not only during SD invasion the recording cortical sites but also when SD occupied distant subcortical/cortical areas. Contralateral cortex not invaded by SD also showed transient depression of gamma activity in awake animals. Introduction of general anesthesia modified the pattern of SD-induced depression: SD evoked the strongest cessation of slow delta activity, milder suppression of fast oscillations and no distant changes in gamma activity., Conclusion: Slow and fast cortical oscillations differ in their vulnerability to SD influence, especially in wakefulness. In the conscious brain, SD produces stronger and spatially broader depression of fast cortical oscillations than slow ones. The frequency-specific effects of SD on cortical activity of awake brain may underlie some previously unexplained clinical features of migraine aura., (© 2024. The Author(s).)

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.

The complexities of translational strategies make this stage of implementing genetic information one of the most challenging to comprehend and, simultaneously, perhaps the most engaging. It is evident that this diverse range of strategies results not only from a long evolutionary history, but is also of paramount importance for refining gene expression and metabolic modulation. This notion is particularly accurate for organisms that predominantly exhibit biochemical and physiological reactions with a lack of behavioural ones. Plants are a group of organisms that exhibit such features. Addressing unfavourable environmental conditions plays a pivotal role in plant physiology. This is particularly evident with the changing conditions of global warming and the irrevocable loss or depletion of natural ecosystems. In conceptual terms, the plant response to abiotic stress comprises a set of elaborate and intricate strategies. This is influenced by a range of abiotic factors that cause stressful conditions, and molecular genetic mechanisms that fine-tune metabolic pathways allowing the plant organism to overcome non-standard and non-optimal conditions. This review aims to focus on the current state of the art in the field of translational regulation in plants under abiotic stress conditions. Different regulatory elements and patterns are being assessed chronologically. We deem it important to focus on significant high-performance techniques for studying the genetic information dynamics during the translation phase.

It is well known that neonatal pro-inflammatory challenge (NPC) acquire a predisposition to the development of a number of neuropsychiatric diseases: depression, anxiety disorders, autism, attention deficit hyperactivity disorder. Symptoms of these diseases can manifest themselves in adulthood and adolescent after repeated exposure to negative influences. Preventing the development of the negative consequences of NPC is one of the main tasks for researchers. The exposure to an enriched environment (EE) was shown to have anxiolytic, anti-depressive, and pro-cognitive effects. The present work was aimed to investigate the effects of the long-term EE on anxious-depressive and conditioned fear behavior in normal male and female rats and subjected to NPC. The NPC was induced by subcutaneous administration of lipopolysaccharide (LPS, 50 μg/kg) on 3d and 5th PNDs. The control animals received saline (SAL). The rats were placed in the EE from 25 to 120 PND. Animals housed in the standard conditions (STAND) served as controls. In adult female and male rats of the STAND groups, LPS did not affect the anxiety, depressive-like behavior and conditioned fear. The EE increased motor and search activity in males and females. In the open field, the EE reduced anxiety in males of the SAL and LPS groups and in females of SAL groups compared to the STAND housed animals. In the elevated plus maze, the EE decreased anxiety only in males of the SAL group. In the sucrose preference test, the EE did not change sucrose consumption in males and females of SAL and LPS groups, while, in the forced swimming test, the EE reduced depressive-like behavior in females of both SAL and LPS groups. The enrichment decreased the contextual conditioned fear in male and female of SAL groups, but not of the LPS group, and did not affect the cue conditioned fear. The corticosterone reactivity to the forced swimming stress increased in males of the EE groups. The basal level of IL-1beta in blood serum decreased in males of the SAL-EE group. Thus, the EE reduced anxiety in males, depressive-like behavior in females, and contextual conditioned fear in males and females compared to the STAND housed animals. Although the NPC did not affect these behaviors in the STAND groups, LPS prevented the beneficial EE effects on anxiety and conditioned fear. The opposing effects of LPS were dependent on sex and type of testing., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

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.

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.

Background: RNA-seq has become a standard technology to quantify mRNA. The measured values usually vary by several orders of magnitude, and while the detection of differences at high values is statistically well grounded, the significance of the differences for rare mRNAs can be weakened by the presence of biological and technical noise., Results: We have developed a method for cleaning RNA-seq data, which improves the detection of differentially expressed genes and specifically genes with low to moderate transcription. Using a data modeling approach, parameters of randomly distributed mRNA counts are identified and reads, most probably originating from technical noise, are removed. We demonstrate that the removal of this random component leads to the significant increase in the number of detected differentially expressed genes, more significant pvalues and no bias towards low-count genes., Conclusion: Application of RNAdeNoise to our RNA-seq data on polysome profiling and several published RNA-seq datasets reveals its suitability for different organisms and sequencing technologies such as Illumina and BGI, shows improved detection of differentially expressed genes, and excludes the subjective setting of thresholds for minimal RNA counts. The program, RNA-seq data, resulted gene lists and examples of use are in the supplementary data and at https://github.com/Deyneko/RNAdeNoise ., (© 2022. The Author(s).)

A broad and amazingly intricate network of mechanisms underlying the decoding of a plant genome into the proteome forces the researcher to design new strategies to enhance both the accumulation of recombinant proteins and their purification from plants and to improve the available relevant strategies. In this paper, we propose new approaches to optimize a codon composition of target genes (case study of interferon-αA) and to search for regulatory sequences (case study of 5'UTR), and we demonstrated their effectiveness in increasing the synthesis of recombinant proteins in plant systems. In addition, we convincingly show that the approach utilizing stabilization of the protein product according to the N-end rule or a new protein-stabilizing partner (thermostable lichenase) is sufficiently effective and results in a significant increase in the protein yield manufactured in a plant system. Moreover, it is validly demonstrated that thermostable lichenase as a protein-stabilizing partner not only has no negative effect on the target protein activity (interferon-αA) integrated in its sequence, but rather enhances the accumulation of the target protein product in plant cells. In addition, the retention of lichenase enzyme activity and interferon biological activity after the incubation of plant protein lysates at 65 °C and precipitation of nontarget proteins with ethanol is applicable to a rapid and inexpensive purification of fusion proteins, thereby confirming the utility of thermostable lichenase as a protein-stabilizing partner for plant systems.

Spreading depolarization (SD), a self-propagated wave of transient depolarization, regularly occurs in the cortex after acute brain insults and is now referred as an important diagnostic and therapeutic target in patients with acute brain injury. Here, we show that the amygdala, the limbic structure responsible for post-injury neuropsychological symptoms, exhibits strong regional heterogeneity in vulnerability to SD with high susceptibility of its basolateral (BLA) region and resilience of its centromedial (CMA) region to triggering SD by acute focal damage. The BLA micro-injury elicited SD twice as often compared with identical injury of the CMA region (71% vs. 33%). Spatiotemporal features of SDs triggered in the amygdala indicated diverse patterns of the SD propagation to the cortex. Our results suggest that even relatively small cerebral structures can exhibit regional gradients in their susceptibility to SD and the heterogeneity may contribute to the generation of complex SD patterns in the injured brain.

Background: For the needs of modern biotechnology, a quantitative approach to the control of regulatory elements at all stages of gene expression has long become indispensable. Such a control regime is impossible without a quantitative analysis of the role of each regulatory element or pattern used. Therefore, it seems important to modify and develop the accuracy, reproducibility, and availability of methods for quantifying the contribution of each regulatory code to the implementation of genetic information., Results: A new vector system for transient expression in plants is described; this system is intended for quantitative analysis of the contribution of regulatory elements to transcription and translation efficiencies. The proposed vector comprises two expression cassettes carrying reporter genes (of the Clostridium thermocellum thermostable lichenase and E. coli β-glucuronidase) under the control of different promoters. Herewith we also propose a new method for quantification of the effect of tested regulatory elements on expression, which relies on assessment of the enzyme activities of reporter proteins taking into account the transcription of their genes., Conclusions: In our view, this approach makes it possible to precisely determine the amounts of reporter proteins and their transcripts at all stages of expression. The efficiency of the proposed system has been validated by the analysis of the roles of known translation enhancers at the stages of transcription and translation., (© 2022. The Author(s).)

Instructors can foster equitable learning environments when they communicate that they value growth and diversity and by providing opportunities for students to reflect and to engage in cross-group interactions with diverse others. Additionally, perspective-taking activities engage empathy and have been used to reduce racial bias. An activity was introduced at the start of the semester to promote a more scientifically informed view of traits and intelligence, while also encouraging creativity, evidence-based thinking, and teamwork in an introductory biology course that was taught in an online synchronous format due to the COVID pandemic. Student groups worked in breakout rooms to create a slide in a shared file describing a research plan to test for human intelligence from the perspective of an alien life form. Students had the freedom to choose their alien to have whatever abilities and intelligence they wanted. The activity was highly student-centered, with students showing through their work that an understanding of intelligence is closely linked to the methods used to measure it. In this way, the activity promoted a more nuanced understanding of traits. This COVID-era invention resulted in a successful strategy that can be used in a variety of course delivery formats to support the teaching of content, as well as to promote a growth learning mindset and inclusive education., Competing Interests: The author declares no conflict of interest., (Copyright © 2022 Pavlova.)

Late blight is one of the most economically important diseases affecting potato and causing a significant loss in yield. The development of transgenic potato plants with enhanced resistance to infection by Phytophthora infestans may represent a possible approach to solving this issue. A comparative study of the leaf response in control potato plants ( S.tuberosum L. cultivar Skoroplodnyi), control transgenic plants expressing the reporter gene of thermostable lichenase (transgenic licB M3 line) and transgenic plants expressing cyanobacterial hybrid genes ∆9-acyl-lipid desaturase (transgenic desC lines) and ∆12-acyl-lipid desaturase (transgenic desA lines) to infection with P. infestans has been performed. The expression of desaturase genes in potato plants enhanced their tolerance to potato late blight agents as compared with the control. The lipid peroxidation level raised in the leaves of the control and transgenic desA plants on third day after inoculation with P. infestans zoospores and remained the same in the transgenic desC plants. The number of total phenolic compounds was increased as early as on the second day after infection in all studied variants and continued to remain the same, except for transgenic desC plants. Accumulation of flavonoids, the main components of the potato leaf phenolic complex, raised on the second day in all studied variants, remained unchanged on the third day in the control plants and decreased in most transgenic plants expressing desaturase genes. The results obtained in our study demonstrate that the expression of genes of Δ9- and Δ12-acyl-lipid desaturases in potato plants enhanced their resistance to P. infestans as compared with the control non-transgenic plants due to concomitant accumulation of phenolic compounds, including flavonoids, in the leaves. All these changes were more pronounced in transgenic desC plants, which indicates that the Δ9-acyllipid desaturase gene appears to be a potential inducer of the production of biological antioxidants in plant cells.

Spreading depolarization (SD), a self-propagating wave of near-complete breakdown of the transmembrane ion gradients with water influx, regularly occurs in traumatized human brain. Here, we investigated long-term neurobehavioral consequences of injury-triggered SDs. Recently, we revealed that SD is reliably triggered by micro-injury of the amygdala, a key brain structure involved in fear processing. Using the classical Pavlovian fear conditioning paradigm, we investigated effects of the post-retrieval amygdala micro-injury and associated SD on fear memory in rats. We found that neither SD nor micro-injury alone affect fear response 24 h later but profoundly change it in subsequent extinction phase. If bilateral injury of the amygdala did not induce SD, fear extinction was severely impaired, while conditioned fear in rats with the identical amygdala injury triggering SD was successfully extinguished similarly to naïve rats. Our study provides first experimental evidence for involvement of injury-induced SD in extinction of traumatic fear memory., (Copyright © 2021 Elsevier B.V. All rights reserved.)

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., (© Springer Nature Switzerland AG 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

As part of a wider reform to scaffold quantitative and research skills throughout the biology major, we introduced course-based undergraduate research experiences (CURE) in sections of a large-enrollment introductory biology laboratory course in a mid-level, public, minority-serving institution. This initiative was undertaken as part of the in the National Science Foundation / Council for Undergraduate Research Transformations Project. Student teams performed two or three experiments, depending on semester. They designed, implemented, analyzed, revised and iterated, wrote scientific paper-style reports, and gave oral presentations. We tested the impact of CURE on student proficiency in experimental design and statistical reasoning, and student research confidence and attitudes over two semesters. We found that students in the CURE sections met the reformed learning objectives for experimental design and statistical reasoning. CURE students also showed higher levels of experimental design proficiency, research self-efficacy, and more expert-like scientific mindsets compared to students in a matched cohort with the traditional design. While students in both groups described labs as a positive experience in end-of-semester reflections, the CURE group showed a high level of engagement with the research process. Students in CURE sections identified components of the research process that were difficult, while also reporting enjoying and valuing research. This study demonstrates improved learning, confidence, and attitudes toward research in a challenging CURE laboratory course where students had significant autonomy combined with appropriate support at a diverse public university., Competing Interests: DR is a member of the editorial board of PLOS One. The remaining authors have declared no competing interests.

Scientific education provides a set of tools to make sense of a complex world by teasing out complicated cause-and-effect relationships, such as the elimination of effects of confounding factors in controlled experiments. There is evidence that depth of understanding of controlled experiments is lacking among undergraduate science students despite exposure to controlled experiments in courses. To examine the understanding of controlled experiments, we developed a two-tiered assessment that includes closed-ended and open-ended questions, with three types of questions, i.e., (i) a scientific scenario about a flawed drug study, (ii) an everyday-life scenario about flawed thinking regarding product effectiveness, and (iii) a direct question about explaining controlled experiments. Consistent with previous findings, we demonstrated that large percentages of students in introductory biology courses at both a research-intensive institution and a primarily undergraduate, minority-serving institution failed to recognize the need to account for confounds. Based on these findings, we tested the hypothesis that scientific literacy could be improved through a course-based intervention using an active learning framework focused on science as a process of evaluating alternative hypotheses. We found start-to-end-of-semester improvement in students' identification of unaccounted confounds with a scientific scenario in an intervention course but not in the control course. Interestingly, students in both the control and intervention courses showed improvement when tested with a scenario based on everyday life. The study findings suggest that a concerning number of college students may not account sufficiently for uncontrolled variables in real-life situations, and we present a widely applicable instructional strategy that improves on this broadly relevant scientific reasoning skill., (Copyright © 2021 Kreher et al.)

Background: Since the beginning of the use of reporter proteins for expression analysis, a variety of approaches have been developed and proposed; both qualitative and quantitative. The lack of simple methods for direct observation of gene expression in living organisms makes it necessary to continue to propose new methods. In this work, we consider a method for the quantitative analysis of the expression of thermostable lichenase from Clostridium thermocellum used as a sensitive reporter protein., Results: In this study, we report the design a high throughput fluorometric method for quantification of thermostable lichenase C. thermocellum using Congo red and further experimental verification of its relevance and efficiency in assessment of the functional role of regulatory sequences in the plant cell., Conclusions: The specific interaction between the dye Congo red and [Formula: see text]-D-glucans formed the background for designing a high-throughput fluorometric assay for quantification of C. thermocellum thermostable lichenase as a reporter protein for plants. This assay (i) makes it possible to precisely measure the amount of reporter protein in a plant sample; (ii) has shown a high sensitivity for quantification of thermostable lichenase; (iii) is more time- and cost-efficient as compared with the Somogyi-Nelson assay; and (iv) is to the least degree dependent on the presence of the tested buffer components as compared with the Somogyi-Nelson assay., (© 2021. The Author(s).)

A large data array on plant gene expression accumulated thanks to comparative omic studies directs the efforts of researchers to the specific or fine effects of the target gene functions and, as a consequence, elaboration of relatively simple and concurrently effective approaches allowing for the insight into the physiological role of gene products. Numerous studies have convincingly demonstrated the efficacy of transient expression strategy for characterization of the plant gene functions. The review goals are (i) to consider the advantages and limitations of different plant systems and methods of transient expression used to find out the role of gene products; (ii) to summarize the current data on the use of the transient expression approaches for the insight into fine mechanisms underlying the gene function; and (iii) to outline the accomplishments in efficient transient expression of plant genes. In general, the review discusses the main and critical steps in each of the methods of transient gene expression in plants; areas of their application; main results obtained using plant objects; their contribution to our knowledge about the fine mechanisms of the plant gene functions underlying plant growth and development; and clarification of the mechanisms regulating complex metabolic pathways.

Objectives : Spreading depolarization (SD) is a well-recognized component of the stress response of the cortex to its acute injury. Cortical SD has been shown to occur in severe brain insults and standard neurosurgical procedures in patients and is supposed to promote delayed secondary brain injuries. Stereotactic surgery and site-specific intracerebral microinjections produce a small tissue injury when a thin needle is inserted directly into the brain parenchyma (via the cannula guide). The present study was designed to examine whether such a parenchymal damage can trigger SD. Methods : Experiments were performed in awake freely moving rats with simultaneous video-monitoring of behavior and recording of SD-related DC potentials in the cortex and striatum. A parenchymal damage was produced by 1-mm protruding of thin (0.3-mm diameter) cannula beyond the tip of cannula guide preliminary implanted into the amygdala or deep cortical layers. Results : We found that the micro-injury of the brain parenchyma the volume of which did not exceed 0.3 mm 3 was sufficient to initiate SD in a very high proportion of rats (75-100%). The amygdala showed increased resistance against the injury-induced SD compared to the cortex. We further showed that SD triggered by the local micro-injury invaded remote intact regions of the cortico-striatal system and evoked specific changes in spontaneous animal behavior. Discussion : The findings indicate that SD may represent a previously unidentified side effect of local parenchymal injury during site-specific microinjections and stereotactic surgery.

Clostridium thermocellum lichenase (endo-β-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.

The control of translation in the course of gene expression regulation plays a crucial role in plants' cellular events and, particularly, in responses to environmental factors. The paradox of the great variance between levels of mRNAs and their protein products in eukaryotic cells, including plants, requires thorough investigation of the regulatory mechanisms of translation. A wide and amazingly complex network of mechanisms decoding the plant genome into proteome challenges researchers to design new methods for genome-wide analysis of translational control, develop computational algorithms detecting regulatory mRNA contexts, and to establish rules underlying differential translation. The aims of this review are to (i) describe the experimental approaches for investigation of differential translation in plants on a genome-wide scale; (ii) summarize the current data on computational algorithms for detection of specific structure⁻function features and key determinants in plant mRNAs and their correlation with translation efficiency; (iii) highlight the methods for experimental verification of existed and theoretically predicted features within plant mRNAs important for their differential translation; and finally (iv) to discuss the perspectives of discovering the specific structural features of plant mRNA that mediate differential translation control by the combination of computational and experimental approaches.

The main specific features of β-1,3-1,4-glucanases (or lichenases, EC 3.2.1.73), the enzymes that in a strictly specific manner hydrolyze β-glucans of many cereal species and lichens containing β-1,3 and β-1,4 bonds, are reviewed as well as the current strategies used for their protein design, which have been successfully applied to make lichenases more attractive and promising for biocatalytic conversion of biomass, in particular, in the areas of their biotechnological application, such as brewing industry, animal feed manufacture, and biofuel production, which will in future allow these technologies to be economically and ecologically beneficial.

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.

Recombinant interferon-β1b (IFN-β1b) is an effective remedy against multiple sclerosis and other diseases. However, use of small polypeptide (molecular weight is around 18.5 kDa) is limited due to poor solubility, stability, and short half-life in systemic circulation. To solve this problem, we constructed two variants of PASylated IFN-β1b, with PAS sequence at C- or N-terminus of IFN-β1b. The PAS-modified proteins demonstrated 4-fold increase in hydrodynamic volume of the molecule combined with 2-fold increase of in vitro biological activity, as well as advanced stability and solubility of the protein in solution as opposed to unmodified IFN-β1b. Our results demonstrate that PASylation has a positive impact on stability, solubility, and functional activity of IFN-β1b and potentially might improve pharmacokinetic properties of the molecule as a therapeutic agent.

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.

Laccases, blue copper-containing oxidases, ≿ an play an important role in a variety of natural processes. The majority of fungal laccases are encoded by multigene families that express closely related proteins with distinct functions. Currently, only the properties of major gene products of the fungal laccase families have been described. Our study is focused on identification and characterization of laccase genes, which are transcribed in basidiomycete Trametes hirsuta 072, an efficient lignin degrader, in a liquid medium, both without and with induction of laccase transcription by copper ions. We carried out production of cDNA libraries from total fungal RNA, followed by suppression subtractive hybridization and mirror orientation selection procedures, and then used Next Generation Sequencing to identify low abundance and differentially expressed laccase transcripts. This approach resulted in description of five laccase genes of the fungal family, which, according to the phylogenetic analysis, belong to distinct clusters within the Trametes genus. Further analysis established similarity of physical, chemical, and catalytic properties between laccases inside each cluster. Structural modeling suggested importance of the sequence differences in the clusters for laccase substrate specificity and catalytic efficiency. The implications of the laccase variations for the fungal physiology are discussed., (Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)

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.

The Clostridium thermocellum lichenase (endo-β-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., (Copyright © 2014 Elsevier B.V. All rights reserved.)

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.

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.

The influence of GABA(A) receptor agonist (muscimol hydrobromide, 0.1 μg/0.5 μL) and antagonist (bicuculline methiodide, 0.2 μg/0.5 μL) injections into the right and the left basolateral amygdala on the behavior of the high- and low-anxiety rats was investigated in elevated plus-maze test. High- and low-anxiety rats had different sensitivities to injections of GABA receptor agents. Administration of muscimol increased open-arm time only in the high-anxiety rats, indicating anxiolytic-like effect. Injection of bicuculline increased aggression of all rats, as well as enhanced locomotion and exploration in high-anxiety rats, increased open-arm time and emotionality in the low-anxiety animals. More powerful changes in behavior of rats were revealed after injections of muscimol into the left amygdala of high-anxiety rats and bicuculline into the right amygdala of low-anxiety ones. The results evidence the existence of individual typological and interhemispheric differences in functioning of the amygdalar GABAergic system.

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.

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.

The active and passive rabbits selected previously on the basis of their behavior in open field and light-dark test, were subjected to fear conditioning using pairing light (4 s) with footshocks (10 Hz, 0.5 s). Heart rate and respiration rate were measured during the classical fear conditioning. Heart rate and respiration rate decreased in response to light before footshock in case of passive-defensive reaction. There were no heart rate and respiration rate reduction in the course of the active defensive reaction. In active rabbits, as compared to passive ones, the frequency of active locomotors reactions and heart rate were higher, the decrease of respiration rate to light was observed at later stages of training, and the detected bradycardia was not stable. Thus, based upon vegetative characteristics, the active rabbits had lower level of fear than passive ones. The active or passive behavioral strategies of animals were preserved during fear conditioning.

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.

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.

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.

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.

By plotting cross-correlation histograms differences were found in interaction of conjectural small "interneurons" and large principal cells of the central and basal amygdalar nuclei in negative emotional situations. The network activity of "nterneurons" was higher than in principal cells. "Interneurons" more frequently had excitatory and inhibitory input or output connections with neighbouring cells, latency of their connections with other cells was smaller than in principal neurons. Interaction of "interneurons" and principal cells differed in animals with active and passive behavioural strategy in negative emotional situations. As compared to active animals, in passive rabbits inhibitory connections to "interneurons" from other cells occurred more frequently, excitatory or inhibitory connections from "interneurons" to principal cells appeared more rare.

The problems of improvement of outpatient care are considered, methodology of more active implementation of hospitalization technologies, the use of day care, hospital at home is proposed. It is emphasized that the introduction of hospitalization technology makes it less than treatment in hospital costs to improve quality of life for huge numbers of patients.

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.

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.

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.

Homologous DNA recombination in eukaryotes is necessary to maintain genome stability and integrity and for correct chromosome segregation and formation of new haplotypes in meiosis. At the same time, genetic determination and nonrandomness of meiotic recombination restrict the introgression of genes and generation of unique genotypes. As one of the approaches to study and induce meiotic recombination in plants, it is recommended to use the recA gene of Escherichia coli. It is shown that the recA and NLS-recA-licBM3 genes have maternal inheritance and are expressed in the progeny of transgenic tomato plants. Plants expressing recA or NLS-recA-licBM3 and containing one T-DNA insertion do not differ in pollen fertility from original nontransgenic forms and can therefore be used for comparative studies of the effect of bacterial recombinases on meiotic recombination between linked genes.

The work is part of a study of the gene pool for Daghestan ethnic groups. In total, 38 alleles and eight genotypes were studied at 14 loci (AB0, Rhesus, P, Lewis, Kell, HP, GC, C'3, TF, 6-PGD, GLO1, ESD, ACP, and PGM1) of immunogenetic and biochemical polymorphic gene systems. A high frequency of allele d of the Rhesus system was observed in all populations examined (0.399-0.474). Among the rare haplotypes of the Rhesus system, we observed CDE in the Degva population, Cde in the Sergokala and Degva populations, and cdE in the Sergokala and Vanashimakhi populations. The typical Caucasian ACP1c allele of the ACP1 locus, which is rather uncommon, was observed at a relatively high frequency in three (Segokala, Vanashimakhi, and Gubden) of the four local populations under study. In the Lewis system, a high frequency of the Le(a+b+) phenotype, which is characteristic of early childhood, was detected in the adult populations of Sergokala and Degva. The rare PGM1v allele of the phosphoglucomutase 1 system (PGM 1) was additionally observed in the Sergokala population. Statistical analysis identified 19 cases where the observed phenotype frequencies significantly differed from the frequencies expected from the Hardy--Weinberg equilibrium.

The interhemisphere interaction of neurons in bilateral derivations from parietal and sensomotor areas of neocortex and the area CA1 of hippocampus were studied in rabbits with active and passive behavioural strategy in the open field by plotting histograms of crosscorrelation. In passive animals, there was asymmetry in bilateral neuronal interaction: with right-sided dominance in the neocortex and with left-sided that--in the hippocampus. On the contrary, in active rabbits, the left-sided dominance was observed in the neocortex, and the lateralization was not revealed in the hippocampus. The brain laterality was reflected in motor asymmetry of animals in preferring left or right turns in the open field. Passive rabbits made relatively more left turns, and the active animals--right turns. Systemic administration of agonist GABA(B) receptors phenibut decreased behavioural responses to emotional stimuli and eliminated interhemisphere asymmetry observed usually in negative emotional situations. Thus the interhemisphere asymmetry of the neocortex and hippocampus is correlated with individual typological characteristics of animals and reflects the readiness to preferential forms of behavioural responses in active and passive rabbits.

Thermostable lichenase encoded by licB gene of Clostridium thermocellum can be used as a reporter protein in plant, bacterial, yeast, and mammalian cells. It has important advantages of high sensitivity and specificity in qualitative and quantitative assays. Deletion variants of LicB (e.g., LicBM3) retain its enzymatic activity and thermostability and can be expressed in translational fusion with target proteins without compromising with their properties. Fusion with the lichenase reporter is especially convenient for the heterologous expression of proteins whose analysis is difficult or compromised by host enzyme activities, as it is in case of fatty acid desaturases occurring in all groups of organisms. Recombinant desaturase-lichenase genes can be used for creating genetically modified (GM) plants with improved chill tolerance. Development of an analytical method for detection of fused desaturase-lichenase transgenes is necessary both for production of GM plants and for their certification. Here, we report a multiplex polymerase chain reaction method for detection of desA and desC desaturase genes of cyanobacteria Synechocystis sp. PCC6803 and Synechococcus vulcanus, respectively, fused to licBM3 reporter in GM plants.

Three genes mutations in which cause familial forms of Alzheimer's disease are known to date:PSEN1, PSEN2 and APP; and APOE gene polymorphism is a strong risk factor for Alzheimer's disease. We have evaluated allele and genotype frequency distribution of rs11136000 polymorphism in clusterin (CLU) gene (or apolipoprotein J, APOJ) in populations of three Russian regions and i nAlzheimhner's diseasepatients. Genome-wideassociation studies in samples from several European populations have recently revealed highly significant association o fCLU gene with AD (p = 8.5 x 10(-10)). We found no differences in allele and genotype frequencies of rs11136000 between populations from Moscow, Ural and Siberia regions. The allele frequencies are close to those in European populations. The genetic association analysis in cohort of Alzheimer's disease patients and normal individuals (>500 individuals ineach group) revealed no significant association of the rs11136000 polymorphism in CLU with Alzheimer's disease in Russian populations. Although our resultsdo not confirm the role of CLU gene as a majorgenetic factor forcommon form of Alzheimer's disease, the data do not rule out the possibility of modest effect of CLU and interaction between CLU and APOE genotypes in etiology of Alzheimer's disease.

Cross-correlation histograms were used to compare the interaction of close-lying cells in the hippocampus (field CA1) and the parietal-temporal areas of the neocortex in active and passive rabbits during exposure to emotionally significant stimuli. Interaction of hippocampal neurons depended on the type of the behavioral response to the stimulus. The greatest changes from baseline were seen in active rabbits in orientational-investigative and active defensive reactions, while the greatest changes in passive animals were in freezing. In all states, cases of common inputs to hippocampal neurons were found more frequently in passive rabbits and excitatory links between neurons with short delays (up to 40 msec) were more frequent in active rabbits. Interaction of neocortical neurons, in contrast with hippocampal neurons, was less dependent on the type of behavioral response to the stimulus and the animal's behavioral strategy. These results provide evidence of individual-typological features in information processing in hippocampal field CA1 in active and passive animals in emotionally negative situations.