Endogenous Brassinosteroids Are Involved in the Formation of Salt Resistance in Plants.

The endogenous brassinosteroid (BS) profile was for the first time shown to change in response to salt stress in potato plants. A group of 6-keto-BSs was identified and found to significantly increase in content during salinization in contrast to other groups of hormones examined. A tenfold reduction in the level of endogenous BSs in mutant Arabidopsis thaliana plants with impaired biosynthesis (det2) (or reception (bri1)) of phytosteroids decreased their salt resistance, as evidenced by a lower efficiency of photochemical processes of photosystem II (PSII) and growth inhibition. The results confirmed the idea that endogenous BSs are involved in the formation of salt resistance in plants.

Polymetallic Stress Changes the Endogenous Status of Brassinosteroids and Reduces the Effectiveness of Photochemical Reactions Photosystem II in Barley Plants.

The effect of polymetallic stress (Mn2+, Cd2+, Cu2+, Ni2+, Zn2+, Pb2+, and Al3+) of different duration and intensity on the endogenous brassinosteroids (BS) content and photochemical activity of photosystem II in barley plants was studied. The content of steroid hormones was determined by a two-stage enzyme immunoassay. It was established for the first time that barley plants responded to polymetallic stress by changing the endogenous content of various phytosteroid groups (24S-methylBS, 24-epiBS, 28-homoBS, B-lactoneBS, and 6-ketoBS). It was shown that the stress-dependent dynamics of changes in the endogenous content of various BS groups is characterized by organ specificity and is determined by the age of plants, the intensity of the acting stressor, and the specificity of brassinosteroids. The observed changes in the endogenous status of BS in barley plants under polymetallic stress are accompanied by inhibition of growth processes, a decrease in the content of the main photosynthetic pigments, and a slight decrease in the efficiency of photochemical processes in PS II (Fv/Fm, ETR, qN, and NPQ). Considering the multifunctional stress-protective effect of brassinosteroids, it is assumed that the change in the endogenous status of BS against the background of polymetallic pollution is adaptive and allows minimizing the damaging effects of toxic heavy metals and aluminum.

Quantitative method for the determination of antibiotic-resistant gut bacterial strains for the early diagnosis of chronic arthritis.

Based on the clinical and microbiological monitoring of two groups of children aged 3 to 17 years with acute (n=78) and chronic (n=46) course of reactive arthritis (ReA), a method for early diagnosis of chronic arthritis was developed by determining the number of antibiotic-resistant coprostrains in patients with ReA, characterized by the absence of the need to isolate a pure culture of the pathogen and its identification; inoculation of faeces at a dilution of 10-5 on solid 1.5% GRM-agar with an antibacterial agents used in the treatment of a particular patient, at a minimum inhibitory concentration in the resistance range, followed by incubation and counting of the colonies of microorganisms grown on the plate. A significant relationship between the number of antibiotic-resistant gut bacterial strains and the course of arthritis (acute, chronic) was revealed, and the borderline value of the number of antibiotic-resistant gut bacterial strains was determined - 5×103 CFU/g, which allows differentiating the acute course from the chronic one: in the acute course< 5×103 CFU/g, with chronic - ≥ 5×103 CFU/g. The method allows, at the stage of completion of anti-inflammatory therapy in the active phase of the disease, to identify a risk group for the development of a chronic course of arthritis among patients with ReA, which can contribute to timely therapeutic measures aimed at preventing recurrence of the disease and making the patient disabled.

Exogenic Melatonin Reduces the Toxic Effect of Polymetallic Stress on Barley Plants.

The effect of melatonin on growth parameters, the photochemical activity of photosystem II (PS II), the content of the main photosynthetic pigments, and lipid peroxidation in barley plant leaves under polymetallic stress were studied. Melatonin reduces the toxic effect of polymetals on biomass accumulation, root growth, and maintenance of the assimilating surface. The protective action of the hormone is based on its ability to reduce the intensity of oxidative stress by maintaining the level of carotenoids and increasing the activity of superoxide dismutase, but not by regulating the photochemical activity of chloroplasts. The effectiveness of melatonin does not depend on the duration of exposure. The data obtained can be the basis for optimizing the use of melatonin as a plant priming inducer.

Melatonin Supports Photochemical Activity of Assimilation Apparatus and Delays Senescence of Leaves of Monocotyledonous Plants.

Melatonin supports the photochemical activity of photosystem II (PS II) and slows down the degradation of the main photosynthetic pigments during aging of leaves of monocotyledonous plants. The protective effect of melatonin is manifested in an increase in the maximum (Fv/Fm) and effective (Y(II)) quantum yield of PS II, in an increase of regulated (Y(NPQ)) and a decrease in unregulated dissipation of excitation energy (Y(NO)). These effects are based on the ability of melatonin to reduce the intensity of oxidative stress by maintaining a high level of carotenoids, which exhibit pronounced antioxidant properties, during aging.

[GUT MICROSYMBIOCENOSIS IN CHILDREN WITH REACTIVE ARTHRITIS].

AIM: To study the state of gut microsymbiocenosis in children with reactive arthritis (RA), with the assessment of biofilm formation (BFF) of microsymbionts and the ability to change cytokine levels (their anticyokine activity) in vitro. MATERIALS AND METHODS: The investigation of gut microsymbiocenosis by means of bacteriological method was conducted in 34 children with RA and 25 relatively healthy 3 - 16 year- old children. Microorganisms were identified with the help of MALDI-TOF mass-spectrometry, anticytokine activity (ACA) of microsymbionts - according to Bukharin O.V et al. (2011), biofilm formation - according to O'Toole G.A., Kolter R. (1998). RESULTS: On the ground of species composition differences of gut microbiota discrimination model was created which allowed to separate the group of children with RA from healthy individuals. Microsymbiocenosis of patients with RA was characterized by increasing number of opportunistic microorganisms (OM) (enterobacteria, clostridia, bacteroides, and Candida), BFF and ACA level. CONCLUSION: The obtained data greatly contribute.to the deciphering of spondylo- arthritis and disclose the role of microbial factor under given pathology. Hypercolonisation of human gut with OM, having pronounced ability to BFF and regulating cytokine level, promotes strengthening of arthritogenic potential and serves as additional marker of arthritis development risk in children.

[Dirofilariasis in the Nizhny Novgorod Region].

The paper presents the results of clinical, epidemiological, and epizootological analyses of local cases of human dirofilariasis in the Nizhny Novgorod Region, which suggest that natural and climatic changes, namely the abnormally hot summer in 2010-2011 and increasing migratory processes among human beings and animals, open up possibilities for forming foci ofdirofilariasis outside habitats of Dirofilaria repens.

Development and validation of a HPLC-MS/MS method for the analysis of fatty acids - in the form of FAME ammonium adducts - in human whole blood and erythrocytes to determine omega-3 index.

Recent scientific studies in the field of health and nutrition have unanimously affirmed the importance of consuming the omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), because of their cardioprotective properties. Fatty acid profiling in erythrocyte membranes allows the omega-3 index, which is a recognized indicator of the risk of developing cardiovascular disease, to be calculated. One consequence of the upward trend in healthy lifestyles and longevity is an increase in the number of studies into the omega-3 index, which requires a reliable method for the quantitative analysis of fatty acids. This article describes the development and validation of a sensitive and reproducible liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the quantitative analysis of 23 fatty acids (in the form of fatty acid methyl esters, FAMEs) in 40 µl of whole blood and erythrocytes. The list of acids includes saturated, omega-9 unsaturated, omega-6 unsaturated and omega-3 unsaturated fatty acids as well as their trans-isomers. The limit of quantitation was 250 ng ml-1 for C12:0, C16:0 and C18:0; and 62.5 ng ml-1 for other FAMEs, including EPA, DHA and trans-isomers of FAME C16:1, C18:1 and C18:2 n-6. Sample preparation for fatty acid (FA) esterification/methylation with boron trifluoride-methanol (BF3) has been optimized. Chromatographic separation has been carried out on a C8 column in gradient mode using a mixture of acetonitrile, isopropanol and water with the addition of 0.1% formic acid and 5 mM ammonium formate. As a result, the problem of separating the cis- and trans-isomers of FAME C16:1, C18:1 and C18:2 n-6 has been solved. The electrospray ionization mass spectrometry (ESI-MS) detection of FAMEs, in the form of ammonium adducts, has been optimized for the first time, which has made the method more sensitive that when the protonated species are used. This method has been applied to 12 samples from healthy subjects that consumed omega-3 supplements and has proven to be a reliable tool for determining the omega-3 index.

[Reference values of clinical donor blood analysis].

Basic hemogram parameters were analyzed in 835 blood donors (421 men and 414 women) aged 18 to 67 years) on an ADVIA 120 hematology analyzer. Reference ranges were calculated for each parameter in relation to their distribution (as X mean +/- 2alpha for normally distributed data and 2.5-97.5 per thousand for others). The findings suggest that the normal range should be extended for platelets, red blood cells, hemoglobin, and packed cell volume as compared with the earlier unified normal values, which must be taken into account when deciding whether a subject may be allowed to donate blood.

[Treatment of contusion of moderate severity in children in outpatient clinics]. / Lechenie detei s ushibom golovnogo mozga srednei stepeni tyazhesti v ambulatornykh usloviyakh.

OBJECTIVE: To determine the efficacy of cortexin in treatment of traumatic brain injuries with contusion of moderate severity in children in an outpatient clinics. MATERIALS AND METHODS: Seventy-four patients, aged 6-13 years, with a traumatic brain injury and a moderate brain contusion were examined. A comprehensive clinical examination, including neurological and ophthalmological examinations, EEG, brain CT scan, testing using a set of experimental psychological techniques, was performed. Children of the main group received standard therapy and cortexin. Children of the control group received similar treatment without cortexin. Re-examination was carried out 30 days after the start of treatment. RESULTS: After the treatment, a positive dynamics was noted in both groups. There were a decrease in the severity of subjective symptoms (p<0,01) and focal neurological symptoms (p <0,001) as well as absence of acute waves to physical activity according to EEG results in the main group compared with the control group. Also, EEG showed that cortical electrogenesis corresponded to the age of the patient, hypertensive/hydrocephalic signs were stopped (p <0,05), positive changes in cognitive functions occurred. CONCLUSION: The study showed the positive dynamics in the recovery of cognitive functions, the normalization of EEG parameters and stopping of hypertension-hydrocephalic symptoms in children with traumatic brain injuries with contusion of moderate severity a month after the start of treatment with cortexin.

[The prediction of the spectrum of biological activity and antimicrobial properties of diaminoazoles]. / Prognozirovanie spektra biologicheskoi aktivnosti i antimikrobnye svoistva diaminoazolov.

The work is devoted to predicting and studying biological properties of N-substituted analogs of 3,5-diamino-1,2,4-thiadiazole, which, in their turn, include in the composition of many drugs that exhibit a wide range of pharmacological actions. For searching of new alternative drugs with an antibacterial activity, but lacking resistance of microorganism strains to them, a computer screening of 2N-alkyl-substituted 5-amino-3-imino-1,2,4-thiadiazolines previously synthesized by us was carried out. The prediction of the spectrum of biological activity, as well as the determination of the probable toxicity of these compounds, was performed using the freely available computer programs PASS, Anti-Bac-Pred, and GUSAR. The study of the antibacterial activity in vitro against gram-positive (Staphylococcus aureus, Staphylococcus saprophyticus, Staphylococcus epidermidis) and gram-negative (Escherichia coli, Pseudomonas aeruginosae) bacterial strains was performed by the disco-diffusion method. Experimental data roughly correspond to the predictions.

[Molecular mechanisms of effects of rosuvastatin on systemic oxidative stress and endogenous inflammation in patients with atherosclerosis].

Aim of the study was to investigate peculiarities of effects of rosuvastatin on the state of oxidative stress and endogenous inflammation in patients with extensive atherosclerosis. Patients with extensive atherosclerosis included into the study (n=46, mean age 56.5 +/- 2.2 years) were distributed to 2 equivalent according to clinico-instrumental data groups. To patients of group 1 (n=24) standard therapy was prescribed (antiaggregants, ACE inhibitors, b-adrenoblockers, and nitrates when indicated), patients of group 2 (n=22) in addition to standard therapy took rosuvastatin (10 mg/day). Investigations included measurement of parameters of serum lipid profile, content of thiol groups of blood serum proteins, activity of enzyme glutathione peroxidase, in vivo oxidation of whole blood serum and HDL, concentration of 3-nitrotirosine, high sensitivity C-reactive protein and interleukin-6, activity of type 2IIA secretory phospholipase A2. It was found that level of 3-nitrotirosine and activity of secretory phospholipase A2 together with high sensitivity C-reactive protein appear to be effective markers of systemic oxidative stress and endogenous inflammation in patients with extensive atherosclerosis. Treatment with rosuvastatin in moderate doses significantly suppressed activity of endogenous inflammation and oxidative stress by way of activation of antioxidant system of plasma, decrease of oxidation of fractions of lipoproteins, suppression of " nitrotirosine " stress, as well as partial inhibition of efficacy of action of secretory phospholipase A2, lowering of content of C-reactive protein and interleukin-6.

A Novel Uncultured Bacterium of the Family Gallionellaceae: Description and Genome Reconstruction Based on the Metagenomic Analysis of Microbial Community in Acid Mine Drainage.

Drainage waters at the metal mining areas often have low pH and high content of dissolved metals due to oxidation of sulfide minerals. Extreme conditions limit microbial diversity in- such ecosystems. A drainage water microbial community (6.5'C, pH 2.65) in an open pit at the Sherlovaya Gora polymetallic open-cast mine (Transbaikal region, Eastern Siberia, Russia) was studied using metagenomic techniques. Metagenome sequencing provided information for taxonomic and functional characterization of the micro- bial community. The majority of microorganisms belonged to a single uncultured lineage representing a new Betaproteobacteria species of the genus Gallionella. While no.acidophiles are known among the cultured members of the family Gallionellaceae, similar 16S rRNA gene sequences were detected in acid mine drain- ages. Bacteria ofthe genera Thiobacillus, Acidobacterium, Acidisphaera, and Acidithiobacillus,-which are com- mon in acid mine drainage environments, were the minor components of the community. Metagenomic data were -used to determine the almost complete (-3.4 Mb) composite genome of the new bacterial. lineage desig- nated Candidatus Gallionella acididurans ShG14-8. Genome analysis revealed that Fe(II) oxidation probably involved the cytochromes localized on the outer membrane of the cell. The electron transport chain included NADH dehydrogenase, a cytochrome bc1 complex, an alternative complex III, and cytochrome oxidases of the bd, cbb3, and bo3 types. Oxidation of reduced sulfur compounds probably involved the Sox system, sul- fide-quinone oxidoreductase, adenyl sulfate reductase, and sulfate adenyltransferase. The genes required for autotrophic carbon assimilation via the Calvin cycle were present, while no pathway for nitrogen fixation was revealed. High numbers of RND metal transporters and P type ATPases were probably responsible for resis- tance to heavy metals. The new microorganism was an aerobic chemolithoautotroph of the group of psychrotolerant iron- and sulfur-oxidizing acidophiles of the family Gallionellaceae, which are common in acid mine drainages.

Phylogenetic and Functional Prokaryotic Diversity in the Hoito-Gol Mesothermal Mineral Spring (Eastern Savan Mountains. Buryat Renuhlie).

High-throughput sequencing was used for comparative analysis of microbial communities of the water and mat from the Hoito-Gol mesothermal mineral sulfide spring (Eastern Sayan Mountains, Buryat Republic). Activity of microbial communities was determined. While both spring biotopes were dominated by members of three bacterial phyla, Proteobacteria, Bacteroidetes, and Firmicutes, they differed drastically in the composition of predominant phylotypes (at the genus level). In the water, the organisms wide spread in aquatic'environments were predominant, mostly aerobic chemoorganotrophs of the generaAcinetobacter, Pe- dobacter, and Flavobacterium. In the microbial mat,;the organisms actively involved in the sulfur cycle predominated, including sulfur-reducing bacteria Sulfurospirillum, sulfate-reducing deltaproteobacteria, sulfur- oxidizing chemoautotrophic bacteria, anoxygenic phototrophic bacteria of,the phyla Chloroflexi and Chloro- bi, as well as purple bacteria belonging to the Q-, P--, and y-Proteobacteria. Microbial mats of the spring exhibited higher phylogenetic diversity compared to high-temperature mats containing photosynthetic microorganisms.

A role for CCL28-CCR3 in T-cell homing to the human upper airway mucosa.

Lymphocyte recruitment to peripheral tissues is fundamental for immune surveillance and homeostasis, but the chemokines and chemokine receptors responsible for tissue-specific homing of T cells to the upper airway mucosa have not been determined. To address this, we analyzed the chemokines expressed in the normal human nasal mucosa and found that CCL28 is preferentially expressed at a high level on the lumenal face of vascular endothelial cells in the mucosa. Analysis of the cognate chemokine receptors revealed that close to 50% of the CD4(+) T cells in the human nasal mucosa expressed the CCL28 receptor CCR3, whereas CCR3 was hardly detectable on T cells in the small intestine and skin. In the circulation, CCR3(+) T cells comprised a small subset that did not express homing receptors to the intestine or skin. Moreover, depletion of CCR3(+)CD4(+) T cells abrogated the proliferative response of human blood CD4(+) T cells against the opportunistic nasopharyngeal pathogen Haemophilus influenzae, indicating that the CCR3(+)CD4(+) T-cell subset in the circulation contains antigen specificities relevant for the upper airways. Together, these findings indicate that CCL28-CCR3 interactions are involved in the homeostatic trafficking of CD4(+) T cells to the upper airways.

Thiadiazole-derived expanded heteroazaporphyrinoids.

[structure: see text] Heteroannulenes 1 and 3 containing three subunits of isoindole or pyrrole, respectively, three 1,3,4-thiadiazole moieties, and six aza-bridges have been synthesized by reaction of the corresponding diiminoisoindoline or diiminopyrroline with 2,5-diamino-1,3,4-thiadiazole. These conjugated nonaromatic 30 pi-electron macrocycles coordinate three metal ions within their central cavity and constitute one of the rare examples of expanded heteroazaporphyrinoids.

Detection of diphtheria antitoxin by four different methods.

OBJECTIVE: To investigate the reliability of the different methods used in Norway and Russia for detection of diphtheria antitoxin. METHODS: One hundred and twenty-two sera were selected among Russian serum samples previously collected for seroepidemiologic studies of diphtheria antitoxin. The sera were selected to cover the total antitoxin range and were analyzed by four different antidiphtheria toxin assays: an in vitro toxin neutralization test using Vero cells (in vitro NT), an in vivo neutralization test using rabbit skin inoculation (in vivo NT), an indirect enzyme immunoassay (EIA) and a passive hemagglutination assay (PHA). The results were expressed according to the international standard as: not protected (<0.01 IU/mL), relatively protected (0.01-0.1 IU/mL) or protected (≥0.1 IU/mL). The sensitivity, specificity and inter-rater agreement (K or Kw) of each method were related to the in vitro NT selected as the reference method. RESULTS: The in vivo NT test corresponded very well with the in vitro NT in its ability to differentiate between protection/relative protection and no protection (sensitivity 97%, specificity 87% and K=0.84). The EIA test showed a high sensitivity (96%), but since many sera were categorized as protected rather than not protected, the specificity (30%) and inter-rater agreement (K=0.29) were low. The PHA test had a very high specificity (100%) but a low sensitivity (86%). CONCLUSIONS: The agreement between the two neutralization tests was high. If none of the neutralization assays is routinely available, the PHA test can be used to predict the need for vaccination on an individual basis but should not be used for seroepidemiologic studies, since the protection rate for diphtheria would be falsely too low, due to the lower sensitivity. The indirect EIA test used in this study should not be used routinely.

Mapping using human blood composition data.

The mapping of elemental composition using INAA on the base of whole blood and hair of inhabitants (2500 samples) was made in Uzbekistan (CIS). The average concentrations of 15 elements (24 for hair) were determined. The results obtained were compared to regional medical statistics. The correlations for various diseases were obtained. The maps of human blood composition in comparison to those for human hair composition seem to be less significant in terms of regional contamination (probably because of stronger homeostatic control of blood). On the other hand, specific changes of blood composition were detected for some occupational groups. Relationships of blood and hair elemental composition and their relative significance are discussed.

Mapping of ecologically unfavorable territories based on human hair composition.

As was shown (1), analysis of human hair on the population level and mapping of large territories using hair elemental composition are promising approaches for estimation of both the environmental situation and the population health status. In (1,2) the map of Uzbekistan (sampling in 1981) was discussed. Ten years later (1991), samples from the territory in the vicinity of the drying out Aral Sea were taken again. Samples were analyzed for 24 elements using instrumental neutron activation analysis. Comparison of the data and maps drawn for 1981 and 1991 and their comparison with changes of the health status have shown that repeated mapping of territories using data on human hair elemental composition could be used in medical geography, especially for prediction of health status changes in ecologically unfavorable areas.

Electrochemical oxidation of drug residues in water by the example of tetracycline, gentamicine and aspirin.

Electro-chemical oxidation as a method to destroy drug residues like aspirin, tetracycline or gentamicine in water was investigated with C-anodes (modified by manganese oxides) and Pt anodes. The mechanism of aspirin and tetracycline oxidation and the influence of the biocide effect was observed using GC-MS and three different microbiological tests. In general, the biological availability increases with progressive oxidation of the antibiotics.