Characteristics of the Airway Microbiome of Cystic Fibrosis Patients.

Microbiota as an integral component of human body is actively investigated, including by massively parallel sequencing. However, microbiomes of lungs and sinuses have become the object of scientific attention only in the last decade. For patients with cystic fibrosis, monitoring the state of respiratory tract microorganisms is essential for maintaining lung function. Here, we studied the role of sinuses and polyps in the formation of respiratory tract microbiome. We identified Proteobacteria in the sinuses and samples from the lower respiratory tract (even in childhood). In some cases, they were accompanied by potentially dangerous basidiomycetes. The presence of polyps did not affect formation of the sinus microbiome. Proteobacteria are decisive in reducing the biodiversity of lung and sinus microbiomes, which correlated with the worsening of the lung function indicators. Soft mutations in the CFTR gene contribute to the formation of safer microbiome even in heterozygotes with class I mutations.

Depressive Behavior and Monoamine Contents in Brain Structures of Rats During Chronic Overcrowding.

Effects of chronic overcrowding on the development of depressive-like behavior and changes in monoamine contents in the brain structures involved in the development of depression were studied in rats. It was shown that overcrowding increased grooming duration and did not change the number of crossed squares and postures in the open-field test. In the forced swimming test, overcrowding increased immobility time and deceased the duration of the first episode of active swimming, which attested to depressive-like behavior. Overcrowding reduced the content of dopamine in the striatum, its metabolites (homovanilic and dioxyphenylacetic acids) and dopamine metabolism rate in the hypothalamus, and increased the concentrations of norepinephrine, homovanilic acid, and dioxyphenylacetic acid in the olfactory bulb and homovanilic acid in the prefrontal cortex. No changes were found in the nucleus accumbens. Serotonin content did not change in all analyzed structures. It was hypothesized that the observed changes in the content of monoamines and their metabolites in certain brain structures could underlie the depressive-like behavior induced by overcrowding in rats.

Immunological parameters of the blood and monoamine content in the brain of rats during long-term overcrowding.

Blood immunological parameters (cytokine profile and interferon status) and the level of monoamines and their metabolites in various brain structures (amygdala, hippocampus, septum, and hypothalamus) were studied in rats kept under standard conditions or in overpopulated cages. Long-term overcrowding was associated with reduced expression of IL-4 gene, increased transcription of IL-17, and decreased production of IFN-γ, which attested to impaired humoral and cell-mediated immunity and disturbances in IFN-γ synthesis at the post-transcriptional level. Under these conditions, the levels of norepinephrine and dopamine decreased in the septum, but increased in the hypothalamus. The amount of dopamine metabolite dihydroxyphenylacetic acid decreased in both these structures, and the index of dopamine metabolism (dihydroxyphenylacetic acid/dopamine ratio, DOPAC/dopamine) decreased only in the hypothalamus. Overcrowding was not followed by changes in the parameters of noradrenergic and dopaminergic systems in the amygdala and hippocampus and serotoninergic system in all study structures.

[Detection of c-fos expression in animal brain in a pilocarpine model of temporal lobe epilepsy].

The dynamics of the involvement of different brain structures in a pathological process is very important for decoding the mechanisms of temporal lobe epilepsy. In this work, the experimental model of temporal lobe epilepsy induced by lithium chloride and pilocarpine was used. The method of immunochemical detection of the immediate early gene c-fos was used as an indicator of functioning neurons in the brain. The c-fos expression was determined at different time points (30, 60 and 90 min) after the pilocarpine injection. An increase in the c-fos expression was observed in neuronal populations during the development of the status epilepticus, the time and degree of involvement of different brain structures being different. The expression of c-fos was first observed in the piriform cortex, the olfactory tubercle, thalamic nuclei, lateral habenular nuclei, and the caudate putamen. Then the hippocampus, the septal formation, the amygdala, and basal ganglia were involved in the activation process. In the hypothalamic areas, c-fos expression was observed latest. These data contribute to understanding the mechanisms of temporal lobe epilepsy and searching for the ways of its therapy.

Anxiety level and body weight changes in rats living in overpopulated cages.

Behavioral indicators of anxiety and body weight dynamics were compared in rats kept under standard conditions (4-5 animals per cage) and in overpopulated cages (17 rats per cage). The behavior of rats exposed to overcrowding for a short and long period was evaluated in the open field, light-darkness, and elevated plus-maze tests. The anxiety parameters of animals increased only in the light-darkness test during the acute period of overcrowding and in all tests during chronic overcrowding. Behavioral tests had a negative impact on body weight gain in controls and rats living in overpopulated cages. If the behavioral tests were carried out at the beginning of the experiment, body weight ceased to increase in animals exposed to overcrowding and continued to increase in the controls. Overcrowding without behavioral tests had no effects on the body weight dynamics. The model of chronic (but not acute) overcrowding may be used for evaluation of anxiolytic effects of drugs.

[Effects of human cultural neuronal and mesenchymal stem cells on the rat learning and brain state after acute hypoxia].

The aim of the study was to compare the effects of neurotransplantation of cultural neural stem cells (NSC) and mesenchymal stem cells (MSC) on the rat behaviour and brain state after acute hypoxia. It was shown that development of two-way avoidance defensive conditioning in a shuttle box improved in rats-recipients with NSC, but not MSC as compared to control. Both the transplants of NSC and transplants of MSC exert neuroprotective influence on the rat brain. NSC both in vitro (before transplantation) and in vivo (on day 27 after transplantation) gave rise to all neural cell types: stem/progenitor cells, precursors of neurons and glia, neurons and glial cells. MSC population in vitro and in vivo (on day 10 after transplantation) consisted of fibroblast-like cells which were eliminated by day 20 after transplantation and were surrounded by reactive glia. We suggest that effects of NSC may be connected with their good survival and potential to differentiate into neurons and with trophic influence on the brain of recipient, whereas MSC only have possible positive trophic effect at early stages after transplantation.

[Effects of human and rat interferons-alpha on the behavior of rats of different ages. Comparative study of the homology of amino acid sequences].

Effects of chronic intranasal administration of human and rat interferons alpha on feeding and defensive behavior of rats were studied. Natural leukocyte human interferon "Lokferon" (a mixture of alpha interferon subtypes) and recombinant rat interferon alpha of the first subtype were used in the dose of 350 ME per rat daily. In addition, using the databases NCBI and EBI, we quantitatively estimated homology of amino-acid sequences between different subtypes of human and rat interferons. Both human (mostly in young rats) and rat interferons (mostly in old rats) increased rat feeding behavior after food conditioning to an audio tone. In old (but not in young) rats, both human and rat interferons worsened the ability of time interval assessment. In young (but not old) rats, both interferon kinds improved avoidance conditioning. The degree of homology between different human and rat interferons varied from 72% to 77%. Thus, generally, the effects of rat and human alpha interferons (350 ME) on rat conditioning were similar. This may be due to high degree of homology of amino-acid sequences between the two interferons.

[Dose-dependent effects of neuroimmunomodulator interferon alpha on behavior of humans and animals].

The review analyses the literature on polyfunctional activity of the cytokine interferon-alpha (IA) which has neuroimmunomodulatory properties and is used for treatment of viral and oncologic diseases. The major intention is devoted to action of small and high doses of IA on different forms of animals and human behavior. We suggest that chronic treatment with small IA doses is more effective than high dose therapy.

[The role of interferon-alpha in regulation of nervous system functions].

The review analyses contemporary data on the role of the interferon-alpha in the central nervous systems. Interferon-alpha is one of the key polyfunctional cytokines providing integrative activity of the neuro-immuno-endocrine complex. The emphasis is made on the molecular mechanisms of anti-viral, anti-proliferative and neuromodulating actions of the interferon-alpha in the brain. Mechanisms of its involvement in regulation of pain, sleep, body temperature, circadian rhythms, food consumption etc. are considered. Based on the literature and our data we hypothesized a dose-dependent action of exogenous interferon-alpha on the nervous system. We suggest that optimal schemes of chronic application of small interferon-alpha doses can be more expedient for treatment of viral or oncologic diseases than large doses causing neuropsychiatric disorders.

[Effect of small doses of interferon-alpha on food conditioning in young and ageing rats].

Low doses (10 or 350 ME) of human interferon-alpha (HIA) were intranasally applied to young (3-4 months) and ageing (12-15 months) Wistar rats during food conditioning. In control groups, development of the conditioned reflex to acoustic stimulus (tone) did not differ significantly in young and ageing rats in the course of chronic applications of the HIA. However, the control ageing rats were better than young rats in time-interval conditioning. Small doses of HIA do not cause anorexia in rats whereas large doses do so. Tone-conditioning did not change in rats of both ages when they were treated with 10 ME of the HIA; moreover, 350 ME increased food motivation, especially in young rats. Time-interval conditioning in aging rats was descended by both doses to the level of young rats, whereas in young rats it did not change at all. We suggest that these differences between ages may by accounted for be different affinity and concentration of micro-opiod receptors (which are the targets for the HIA) in the brain structures responsible for food behaviour, and for counting time intervals.

[Influence of small doses human interferon-alpha intranasal injection on behavior of rats of different age].

Behavior of young (3-4 month old) and ageing (12-15 month old) rats was studied during chronic intranasal application of low doses (10 ME or 350 ME) of human interferon-alpha (HIA). In ageing rats HIA did not affect dynamics (days 0th, 8th and 16th) of (a) locomotive and (b) investigative activity in the "open field" test and in two-side defensive conditioning, and (c) decreased anxiety ("open field", "light-darkness" test). In young rats HIA (a) increased locomotive activity by 16th day (it decreased in control), (b) investigative activity did not change (in control it decreased by 8th day; "open field" test), (c) anxiety decreased in the "open field" and increased in "light-darkness" tests, (d) development of conditioned reflex improved (during 2nd learning session in 5 days after the first one). Thus, small doses of HIA differently affected behavior of rats depending on the age and experimental situation. However, both HIA doses changed rats' behavior in the same direction. We suggest that chronic low doses of HIA can regulate different aspects of behavior, but not suppress activity as it is commonly thought. This regulation can be performed via modulation of neuro-immuno-endocrine complex.

[Structural-metabolic changes in neurons of hippocampus and neocortex under the influence of "polydan"].

The aim of this work was to study the influence of different regimens of polydan administration on the structural and metabolic state of neurons of rat hippocampus and neocortex. The animals of experimental groups were administered 75 mg/kg of polydan intraperitoneally once or 5 times. Animals of control groups received similar injections of sodium chloride isotonic solution. Light microscopic study of hippocampus and somatosensory area of neocortex demonstrated 4 types of neurons that could be distinguished according to the degree of their staining with methylene blue. Polydan administration resulted in redistribution of relative proportions of neurons of these types and in increase of the number of nucleoli in these cells, thus indicating an activation of synthetic processes. Quantitative ultrastructural analysis of ribosomes in the cytoplasm of neurons of hippocampal area CA3 and neocortical layer V has confirmed the activation of protein synthesis. Intensification of synthetic processes in neurons was further supported by the analysis of mitochondrial cristae and matrix. It is suggested that synthesis activation in brain neurons is a structural and functional basis for the mnemotropic effect of polydan.

Influences of an acoustic signal with ultrasound components on the acquisition of a defensive conditioned reflex in Wistar rats.

The effects of short (90 sec) exposures to a complex acoustic signal with ultrasound components on the acquisition of a defensive conditioned two-way avoidance reflex using an electric shock as the unconditioned stimulus in a shuttle box were studied in female Wistar rats. This stimulus induced audiogenic convulsions of different severities in 59% of the animals. A scale for assessing the ability of rats to acquire the conditioned two-way avoidance reflex was developed. Presentation of the complex acoustic signal was found to be a powerful stressor for Wistar rats, preventing the acquisition of the reflex in the early stages (four and six days) after presentation. This effect was independent of the presence and severity of audiogenic convulsions in the rats during presentation of the acoustic signal. On repeat training nine days after the acoustic signal (with the first session after four days), acquisition of the reflex was hindered (as compared with controls not presented with the acoustic signal). However, on repeat training at later time points (1.5 months after the complex acoustic signal, with the first session after six days), the rats rapidly achieved the learning criterion (10 correct avoidance responses in a row). On the other hand, if the acoustic signal was presented at different times (immediately or at three or 45 days) after the first training session, the animals' ability to acquire the reflex on repeat training was not impaired at either the early or late periods after exposure to the stressor. These results suggest that the complex acoustic signal impairs short-term memory (the process of acquisition of the conditioned two-way avoidance reflex at the early post-presentation time point) but has no effect on long-term memory or consolidation of the memory trace.

[Influence of acoustic signal with ultrasonic components on the development of defensive conditioned reflex in Wistar rats].

Short-term (90 s) effects of the complex acoustic signal (CAS) with ultrasonic components on the development of defensive conditioned reflex of two-way active avoidance in a shuttle-box were studied in female Wistar rats. The learning ability of rats was measured on a scale designed in our laboratory. It was shown that CAS stimulation triggered an audiogenic seizure of different strength in 59% of animals. The CAS was extremely stressful for Wistar rats: it prevented the active avoidance learning in early terms after its application (the first training session in 4 or 6 days). This effect did not depend on the presence or intensity of audiogenic seizures during CAS. In the second training session in 9 days (the first session was in 4 days), learning was impaired as compared to control without CAS. However, during repeated training procedure 1.5 months after the CAS (the first session in 6 days), rats rapidly reached the criterion of learning (10 consecutive avoidance reactions). On the other hand, if the CAS was presented with different time lags (immediately, in 3 or in 45 days) after the first training session, the ability of animals to learn during the second session was not impaired both in early and late terms after exposure to the stressor. The results suggest that exposure to CAS prevents development of short-term memory but does not affect consolidation process and long-term memory.

A special case of learning disorder in isolant rats as a model of disintegration.

The combination of two factors - isolation and ontogenesis (starting from 21 days) and subsequent training of rats to a cyclical habit - could in some animals (30%) lead to the formation of unusual behavioral strategies consisting of maintenance of the cyclical habit without reinforcement with food. This dissociation from the vital motivation is regarded as an analog of the disintegration phenomenon, a key step in psychoneurotic disorders. The most common type of disturbance in isolated animals was suppression of searching activity, with extreme learning difficulty, though a training process including a two-month break led to a significant increase in motor-searching activity in this situation. These behavioral abnormalities were accompanied by morphological changes in the sensorimotor cortex of the brain, with a relative thinning of layer 5 and selective decreases in the density of satellite glial cells and deviations from the normal correlational relationships between behavioral and neuroglial measures.

Effect of polidan on ultrastructural changes in the cerebral cortex and hippocampus of rats.

Electron microscopy and morphometry of neurons in layer V of the somatosensory area in the neocortex and CA3 field in the dorsal hippocampus showed that single and 5-fold intraperitoneal injection of Polidan was followed by ultrastructural and metabolic changes in neurons reflecting activation of protein synthesis. The number of free ribosomes decreased; the number of polysomes and count of ribosomes in polysomes and tubules of the granular endoplasmic reticulum increased. Study of mitochondria and neuropil showed that Polidan activates synthetic processes in the neocortex and hippocampus. It should be emphasized that single treatment with Polidan led to functional activation of synthetic processes, while 5-fold injection of Polidan was followed by hyperactivation of synthetic processes and depletion of ultrastructures in the neocortex and hippocampus.

[A special case of learning disruption as a model of a disintegration phenomenon].

Combination of two factors in rats such as the isolation (during 2-4 months, beginning from the 21st day of age) and procedure of space cyclic learning results in a rare kind of behavioral strategy (in 30% animals), when the cyclic behavior is realized without food reinforcement. Such a dissociation between the vital motivation and searching behavior may be considered as an analogue of the disintegration phenomenon in neuropsychotic patients. The deep depression of learning owing to lowering of search represents the dominant type of behavioral disorders in isolants. The cyclic habit training, including a 2-months pause between two sessions, significantly increases behavioral search activity. The behaviors are accompanied by morphological shifts in the sensomotor cortex: significant decrease of the fifth layer (giant pyramids) thickness, selective lowering in the satellite glia density and elimination of normal behavior-glia correlations.

The effects of Mexidol on the acquisition of food-related conditioned reflexes and synaptic ultrastructure in field CA1 of the rat hippocampus after single acoustic stimuli with ultrasonic components.

The effects of a complex acoustic signal with ultrasonic components on the ultrastructure of synapses field CA1 of the rat hippocampus were studied in conditions of two-week courses of the wide-spectrum antioxidant Mexidol (compared with an untreated group); the effects of complex acoustic signals on the dynamics of acquisition of a food-related conditioned reflex using a standard stimulus (a tone) and on the acquisition of a trace conditioned reflex to estimating time intervals were also studied, in the same groups of rats. Controls consisted of unstressed rats treated and not treated with Mexidol. Ultrastructural analysis of the redistribution of vesicles in the synaptic terminals of hippocampal field CA1 showed that synaptic transmission was impaired when assessed one day after exposure to the complex acoustic signal. Mexidol prevented impairment of synaptic transmission. The complex acoustic signal had negative effects on conditioned reflex activity in rats and Mexidol had normalizing actions on the acquisition of conditioned reflexes in stressed rats. These results lead to the conclusion that the antioxidant Mexidol can be applied to the prophylaxis of the impairments in CNS cognitive functions frequently seen in stress.

Transplantation of cultured astrocytes attenuates degenerative changes in rats with kainic acid-induced brain damage.

Viability of astrocyte grafts introduced into CA1 pyramidal layer of the left dorsal hippocampus after injection of kainic acid into this brain region and the effects of these grafts on the hippocampus and amygdala were studied on Wistar rats. In rats with astrocyte grafts the degree of destruction in fields CA1-CA2 of the dorsal and ventral hippocampus, fields CA3-CA4 of the ventral hippocampus, and central and basolateral amygdala was lower compared to animals with kainic acid-induced hippocampal damage and control rats; destructions in the dentate fascia were absent. Our results suggest that astrocyte grafts stimulate neurogenesis in the mature brain of recipient rats with kainic acid-induced brain damage.

[Neurogenesis in the mature olfactory bulb and it's possible functional destination]. / Neirogenez v zreloi oboniatel'noi lukovitse i ego vozmozhnoe funktsional'noe prednaznachenie.

This review is concerned with neurogenesis in the mature mammalian brain with emphasis on cell population renewal in the olfactory bulb (OB). The structural and functional features of the OB are considered along with data on neurotropic viruses and toxic dust penetration into the CNS through the OB. We hypothesize a protective role of neurogenesis in the mature OB. This suggests that normal renewal of cell populations in the OB is an important barrier mechanism protecting the brain from invasion of small amounts of harmful neurotropic agents (ex. viruses and particles of toxic dust), which can cause various neurodegenerative diseases.