[Effects of diet on the gut microbiome in patients with depression]. / Vliyanie diety na mikrobiom kishechnika u patsientov s depressiei.

OBJECTIVE: To investigate the effects of diet on the gut microbiota and to assess the relationship of these factors with depression. MATERIAL AND METHODS: Microorganisms that predominate in depressed patients were identified and associations of the identified organisms with the patients' diet were performed. Fourteen depressed patients and 14 healthy volunteers with the same socio-demographic parameters were included in the study. The Hamilton Depression Scale, Generalized Anxiety Disorder Questionnaire, and the Center for Epidemiologic Studies Questionnaire were used. RESULTS: Erysipelatoclostridium and Clostridium innocuum species were 11.3 and 14.4 times higher in depressed patients compared with healthy controls. Fusicatenibacter saccharivorans, Faecalibacterium prausnitzii and Roseburia faecis species, as well as members of the genus Roseburia were statistically significantly more abundant in the healthy volunteers group (6.5, 2.14, 8.75 and 5.2 times more frequently compared to patients). The presence of these microorganisms was correlated with dietary components. CONCLUSION: Our study revealed groups of microorganisms that differ in healthy volunteers and depressed patients. The association of these microorganisms with the diet was shown, which partially confirmed the influence of a «healthy diet¼ on the development of depressive disorders.

Neutrophils in Tissue Trauma of the Skin, Bone, and Lung: Two Sides of the Same Coin.

Following severe tissue injury, patients are exposed to various danger- and microbe-associated molecular patterns, which provoke a strong activation of the neutrophil defense system. Neutrophils trigger and modulate the initial posttraumatic inflammatory response and contribute critically to subsequent repair processes. However, severe trauma can affect central neutrophil functions, including circulation half-life, chemokinesis, phagocytosis, cytokine release, and respiratory burst. Alterations in neutrophil biology may contribute to trauma-associated complications, including immune suppression, sepsis, multiorgan dysfunction, and disturbed tissue regeneration. Furthermore, there is evidence that neutrophil actions depend on the quality of the initial stimulus, including trauma localization and severity, the micromilieu in the affected tissue, and the patient's overall inflammatory status. In the present review, we describe the effects of severe trauma on the neutrophil phenotype and dysfunction and the consequences for tissue repair. We particularly concentrate on the role of neutrophils in wound healing, lung injury, and bone fractures, because these are the most frequently affected tissues in severely injured patients.

GABA production and structure of gadB/gadC genes in Lactobacillus and Bifidobacterium strains from human microbiota.

Gamma-amino butyric acid (GABA) is an active biogenic substance synthesized in plants, fungi, vertebrate animals and bacteria. Lactic acid bacteria are considered the main producers of GABA among bacteria. GABA-producing lactobacilli are isolated from food products such as cheese, yogurt, sourdough, etc. and are the source of bioactive properties assigned to those foods. The ability of human-derived lactobacilli and bifidobacteria to synthesize GABA remains poorly characterized. In this paper, we screened our collection of 135 human-derived Lactobacillus and Bifidobacterium strains for their ability to produce GABA from its precursor monosodium glutamate. Fifty eight strains were able to produce GABA. The most efficient GABA-producers were Bifidobacterium strains (up to 6 g/L). Time profiles of cell growth and GABA production as well as the influence of pyridoxal phosphate on GABA production were studied for L. plantarum 90sk, L. brevis 15f, B. adolescentis 150 and B. angulatum GT102. DNA of these strains was sequenced; the gadB and gadC genes were identified. The presence of these genes was analyzed in 14 metagenomes of healthy individuals. The genes were found in the following genera of bacteria: Bacteroidetes (Bacteroides, Parabacteroides, Alistipes, Odoribacter, Prevotella), Proteobacterium (Esherichia), Firmicutes (Enterococcus), Actinobacteria (Bifidobacterium). These data indicate that gad genes as well as the ability to produce GABA are widely distributed among lactobacilli and bifidobacteria (mainly in L. plantarum, L. brevis, B. adolescentis, B. angulatum, B. dentium) and other gut-derived bacterial species. Perhaps, GABA is involved in the interaction of gut microbiota with the macroorganism and the ability to synthesize GABA may be an important feature in the selection of bacterial strains - psychobiotics.

The crucial role of neutrophil granulocytes in bone fracture healing.

Delayed bone fracture healing and the formation of non-unions represent an important clinical problem, particularly in polytrauma patients who suffer from posttraumatic systemic inflammation. However, the underlying pathomechanisms remain unclear. Neutrophil granulocytes are crucial effector cells in the systemic immune response and represent the most abundant immune cell population in the early fracture haematoma. Here we investigated the role of neutrophils in a mouse model of uncomplicated fracture healing and compromised fracture healing induced by an additional thoracic trauma. Twenty four hours before injury, 50 % of the mice were systemically treated with an anti-Ly-6G-antibody to reduce neutrophil numbers. In the isolated fracture model, Ly-6G-Ab treatment significantly increased the concentration of both pro- and anti-inflammatory cytokines, including interleukin (IL)-6 and IL-10, and chemokines, for example, C-X-C motif ligand 1 (CXCL1) and monocyte chemotactic protein-1 (MCP-1), in the fracture haematoma. Monocyte/macrophage recruitment was also significantly enhanced. After 21 d, bone regeneration was considerably impaired as demonstrated by significantly diminished bone content and impaired mechanical properties of the fracture callus. These results indicate that undisturbed neutrophil recruitment and function in the inflammatory phase after fracture is crucial to initiate downstream responses leading to bone regeneration. In the combined trauma model, the reduction of neutrophil numbers ameliorated pulmonary inflammation but did not provoke any significant effect on bone regeneration, suggesting that neutrophils may not play a crucial pathomechanistic role in compromised fracture healing induced by an additional thoracic trauma.

[Disorders of gut microflora in the pathogenesis of liver cirrhosis and complications of portal hypertension].

A total of 85 patients with alcoholic and viral cirrhosis were included in study to assess the prevalence of dysbiosis and its relationship with the severity of disease, and with development of dyspeptic disorders. Intestinal bacterial over-growth was measured by means of a lactulose breath test, fecal flora was cultured under aerobic and anaerobic conditions. Intestinal bacterial overgrowth and colon dysbiosis were determined in 82.4% of patients with equal prevalence in alcoholic and viral cirrhosis. Intestinal dysbiosis was found to be risk factor of increasing cirrhosis severity and liver dysfunction, as well as development of complications of portal hypertension. It was documented, that intestinal dyspepsia syndrome in cirrhotic patients is strongly associated with the presence of gut microflora disorders.

[Gene therapy in ophthalmology: state of the art]. / Gentherapie in der Ophthalmologie: Stand der Forschung.

The term "gene therapy" denotes the treatment of diseases or gene deficiencies by introduction of genes into cells. To achieve this goal, vectors are used to transfer the genetic information into the cells. Thus, the protein of interest can be overexpressed or silenced. On account of its easy accessibility, the good compartmentalisation and the separation from the main bloodstream by the blood-retina barrier, the eye represents a very attractive target to treat ocular diseases by gene therapy. In this work, we provide an overview of the progress in ocular gene therapy over the last decade and give an outlook on future developments.

[Production of 70 kDa recombinant human heat shock protein in baculovirus expression system and assessment of its antiviral activity].

AIM: To obtain human recombinant 70 kDa heat shock protein (Hsp70) in baculovirus expression system and to study its antiviral activity. MATERIALS AND METHODS: Baculovirus expression system was used to obtain recombinant HSP70. Plasmid pFastBacHTb-Hsp70 containing sequence coding HSP70 gene with insertion of 6 histidine residues in protein reading frame was constructed. Competent cells MAX Efficiency DH 10 Bac were transfected with pFastBacHTb-Hsp70 plasmid with following extraction of recombinant bacmid Bac-Hsp70. In order to obtain baculovirus expressing HSP70, Sf-9 cells were transfected with Bac-Hsp70 bacmid. Hsp70 extraction and purification was performed with column metal-chelating affinity chromatography using Ni2+ ions. Protective efficacy of recombinant human HSP70 was estimated using model of Venezuelan equine encephalitis (VEE) in mice. RESULTS: Recombinant bacmid Bac-Hsp70 was constructed based on Bac-to-Bac expression system. Baculovirus expressing human HSP70 have been produced after transfection of Sf-9 cells with Bac-Hsp70 bacmid. Cultivation of recombinant baculovirus in Sf-9 cells and application of metal-chelating affinity chromatography allowed to extract purified fraction of HSP70. Experiments on mice infected with VEE virus demonstrated significant protection from death after administration of HSP70 in dose 15 mcg/mice. CONCLUSION: Application of baculovirus expression system and insect cell line for accumulation of recombinant baculoviruses in combination with Ni(2+)-mediated metal-chelating affinity chromatography allowed to obtain highly purified human recombinant HSP70 with marked antiviral activity.

[Identification of clinical isolates of Burkholderia cepacia and assessment of their pathogenicity in animal experiments].

In experiments on animals study of pathogenicity of 9 clinical strains of Burkholderia cepacia isolated from patients with chronic lung diseases was performed. Preliminary identification of studied strains by means of biochemical and genetic methods allowed to establish their belonging to B. cepacia species. It was determined that 6 of 9 strains are epidemiologically significant. Experiments showed that bacteria of studied strains are not able to cause infectious process in white mice and hamadryas baboons. Conclusion about appropriateness of development and use of other biological models was made.

[Cholecystostomy in emergent surgery of hepatopancreatobiliary zone organs].

Results of cholecystostomy formation in 26 elderly or senile patients for urgent diseases of hepatopancreatobiliary zone organs and severe coexistent pathology of respiratory organs, cardiovascular system and intoxication, were analyzed. The tactics was proposed and indications for cholecystectomy formation were substantiated.

[The association study of the DRD2 gene C939T polymorphism and schizophrenia.].

Physiological evidences and several association studies suggest that the DRD2 gene is implicated in the pathogenesis of schizophrenia. The DRD2 capital ES, Cyrillic939capital TE, Cyrillic (rs6275) single nucleotide polymorphism was genotyped in 272 patients and 362 healthy controls. An association between this polymorphism and schizophrenia was found (p=0,02), the frequency of the TT genotype being higher in patients compared to the controls (c2=7,2; small er, Cyrillic=0,007; OR=1,96, 95% CI=1,2-3,2). The results obtained replicate the data of a previous study of the association between the T allele and schizophrenia.

[Mutation analysis of functional role of amino acid residues in domain IV of elongation factor G].

Seven variants of elongation factor G (EF-G) from Thermus thermophilus with mutations Glu494Ile, Gly495Asp, Lys496Ile, His509Leu, Lys564Ile and Tyr568Lys located in the beta-sheet of its domain IV and mutation Gly553Asp in a loop between domain III and IV were constructed using polymerase chain reaction. Functional tests demonstrated that only mutation Lys496Ile, located in the vicinity of the loop 501-504, inhibits translocation effectiveness, in the presence of the mutated EF-G. The functional analysis of all mutations constructed up to now in domain IV reveals that only those located in loops 501-504 and 573-578 markedly decrease the translocation activity of EF-G. These loops are located at the tip of domain IV and close to the decoding center of the 30S ribosomal subunit upon EF-G interaction with the ribosome. The functional role of EF-G and its domain IV in ribosomal translocation is discussed.

Contributions of short-lived radioiodines to thyroid doses received by evacuees from the Chernobyl area estimated using early in vivo activity measurements.

A series of in vivo gamma spectrometric measurements of 65 people evacuated from Pripyat 1.5 days after the Chernobyl Nuclear Power Plant Unit 4 explosion was performed in St Petersburg, Russia, as early as 30 April 1986. The historical spectra and interviews were recently processed and the results used for thyroid dose estimation. Activities of 131I in thyroid and 132Te in lungs were determined easily; for estimation of 132I and 133I activities in thyroid, sophisticated methods of spectral processing were developed. According to thyroid measurement data, the mean ratio of 133I/131I activities (at the time of the accident) inhaled by residents of Pripyat was 2.0. The mean ratio of thyroid dose from 133I inhalation to that caused by 131I amounts to 0.3, which confirms the accuracy of dose estimates based on the evolution of the Chernobyl accident. The mean ratio of 132I activity in thyroid to that of 132Te in lungs was assessed from the human measurement data to be 0.2, which is in reasonable agreement with the metabolic properties of these radionuclides. The mean ratio of thyroid dose from 132I originating from 132Te deposited in lungs to the dose caused by 131I was 0.13 +/- 0.02 for Pripyat residents who did not take KI pills and 0.9 +/- 0.1 for persons who took KI pills. Thus, the contribution of short-lived radioiodines to total thyroid dose of Pripyat residents, which was on average 30% for persons who did not use stable iodine prophylaxis, and about 50% for persons who took KI pills on 26-27 April, should be accounted for in the assessment of thyroid health effects.

[Transition between iron spin states in myoglobin: Roentgen absorption]. / Issledovanie perekhoda mezhdu spinovymi sostoianiiami zheleza v mioglobine: analiz rentgenovskogo pogloshcheniia.

The spin transition of Fe in the active center of a myoglobin molecule, stimulated by a temperature variation, was studied by the theoretical multiple scattering analysis of experimental X-ray absorption data. The spin transition was followed by the movement of the Fe ion out of the plane of the heme without substantial changes in the Fe-N distance. It was shown that the X-ray absorption fine structure above the Fe K-edge is sensitive to both local geometry changes near the Fe ion in the active site of the protein and the spin state of the Fe ion itself. The change in the symmetry of Fe coordination lead to modifications of the spectrum shape in the entire interval up to 40 eV above the main edge. It was found that the spin state effects mostly the rising edge, and at energies above 15 eV becomes negligibly small.

Lives renewed: the emergence of a breast cancer survivor movement in Ukraine.

Breast cancer is the leading cause of cancer deaths among women in Ukraine. In addition to the physical toll that breast cancer takes, the psychological and emotional needs of women diagnosed with the disease are great but rarely addressed. Since 1997, with initial technical assistance from the Program for Appropriate Technology in Health (PATH), a nascent breast cancer survivor movement has emerged in Ukraine, which aims to challenge societal stigmatization of the disease. As of mid-2001, survivor groups have been formed in 15 cities, representing 12 of the country's 25 provinces, and groups in eight cities have begun implementing Peer Support Volunteer programmes through which survivors provide information and emotional support to newly diagnosed women in hospital. Survivors also are playing an important role in raising public awareness about the disease through public events, outreach activities and the media. The groups have also forged alliances with key medical professionals specialising in cancer care, both to educate them about the potential role that survivors' groups can play in assisting patients, as well as to enlist their active support for these efforts. Solidifying the groups' organisational structures and strengthening their management and fundraising skills have been vital to success.

The Techa River dosimetry system: methods for the reconstruction of internal dose.

The Mayak Production Association (MPA) was the first facility in the former Soviet Union for the production of plutonium. Significant worker and population exposures occurred as a result of failures in the technological processes in the late 1940's and early 1950's. Residents of many villages downstream on the Techa River were exposed via a variety of pathways; the more significant included drinking of water from the river and external gamma exposure due to proximity to contaminated bottom sediment and shoreline. After the extent of the major contamination of the Techa River became known, several villages on the upper part of the Techa River were evacuated. Organ doses are being reconstructed on the basis of derivation of an historical source term and a simple river model used to simulate the transport of radionuclides downstream and their retention on sediments; measurements of 90Sr content in teeth and the whole body of half of the members of the cohort; and development of the "Techa River Dosimetry System" for computation of the doses.

[Spin-dependent electronic structure near the iron atom in rubredoxin]. / Zavisiashchaia ot spina élektronnaia struktura vblizi atoma zheleza v rubredoksine.

The fine structure of X-ray absorption spectrum of Fe in rubredoxin was interpreted on the basis of the multiple scattering theory and the results of calculations of the self-consistent potential. For biological molecules, such calculations were made for the first time. It was found that the Fe-S interaction is the main factor, which determines the electronic structure of the protein active center. The changes in spectrum shape are mostly due to the spin configuration of 3d-electrons. It was shown that the dipole transition element significantly changes near the absorption edge; therefore, it is impossible to determine the distribution of unoccupied electronic p-states directly from experiment. However, the results of calculations obtained in this work are consistent with the corresponding experimental data, indicating the adequacy of the calculated densities of free electronic states.