Changed regulation of granulocyte NADPH oxidase activity in the mouse model of obesity-induced type 2 diabetes mellitus.

NADPH oxidase is a target of hyperglycemia in type 2 diabetes mellitus (T2DM), which causes dysregulation of enzyme. Alterations in regulation of NADPH oxidase activity mediated receptor and non-receptor signaling in bone marrow granulocytes of mice with obesity-induced T2DM were studied. The animals fed high fat diet (516 kcal/100 g) for 16 weeks. NADPH oxidase-related generation of reactive species (RS) at normo- and hyperthermia was estimated using chemiluminescent analysis. The redox status of the cells was assessed by Redox Sensor Red CC-1. Baseline biochemical indicators in blood (glucose, cholesterol, HDL and LDL levels) were significant higher in T2DM mice versus controls. Using specific inhibitors, signaling mediated by formyl peptide receptors (FPRs) to NADPH oxidase was shown to involve PLC, PKC, cytochrome p450 in both control and T2DM groups and PLA2 in controls. In T2DM regulation of NADPH oxidase activity via mFpr1, a high-affinity receptors, occurred with a significant increase of the role of PKC isoforms and suppression of PLA2 participation. Significant differences between this regulation via mFpr2, low-affinity receptors, were not found. Non-receptor activation of NADPH oxidase with ionomycin (Ca2+ ionophore) or phorbol ester (direct activator of PKC isoforms) did not revealed differences in the kinetic parameters between groups at 37 °C and 40 °C. When these agents were used together (synergistic effect), lower sensitivity of cells to ionophore was observed in T2DM at both temperatures. Redox status in responses to opsonized zymosan was higher in T2DM mice at 37 °C and similar to control levels at 40 °C. ROC-analysis identified Tmax, RS production and effect of opsonized zymosan as the most significant predictors for discriminating between groups. It was concluded that Ca2+-dependent/PKC-mediated regulation of NADPH oxidase activity was altered in BM granulocytes from diabetic mice.

Water Quality, Toxicity and Diversity of Planktonic and Benthic Cyanobacteria in Pristine Ancient Lake Khubsugul (Hövsgöl), Mongolia.

For the first time, microcystin-producing cyanobacteria have been detected in Khubsugul, which is ancient, pristine and one of the world's largest lakes. The microcystin synthetase genes belonged to the genera Nostoc, Microcystis and possibly Snowella spp. No microcystins were found in the water of the lake. Using the HPLC-HRMS/TOF, five microcystin congeners were identified in biofilms from stony substrates sampled in the coastal zone. The concentration of microcystins in biofilms was low: 41.95 µg g-1 d. wt. by ELISA and 55.8 µg g-1 d. wt. using HPLC. The taxonomic composition of planktonic and benthic cyanobacterial communities was determined by means of microscopy and high-throughput sequencing of 16S rDNA amplicons. Nostocales cyanobacteria dominated benthos of Lake Khubsugul and Synechococcales-plankton. The abundance of cyanobacteria was low both in plankton and benthos; there was no mass development of cyanobacteria. Hydrochemical and microbiological analyses showed that the water in the lake was clean; the number of faecal microorganisms was significantly below the acceptable guideline values. Hydrochemical and hydrophysical parameters, and the concentration of chlorophyll a, were low and within the range of values recorded in the 1970s to 1990s, and corresponded to the oligotrophic state of the lake. There were no signs of anthropogenic eutrophication of the lake and no conditions for the cyanobacterial blooms.

Selenium Nanoparticles Can Influence the Immune Response Due to Interactions with Antibodies and Modulation of the Physiological State of Granulocytes.

Currently, selenium nanoparticles (SeNPs) are considered potential immunomodulatory agents and as targets for activity modulation are granulocytes, which have the most abundant population of immune blood cells. The present study aims to evaluate the cytotoxic effect and its effect on the functional responses of granulocytes. In addition to the intrinsic activity of SeNPs, we studied the activity of the combination of SeNPs and IgG antibodies. Using laser ablation and fragmentation, we obtained nanoparticles with an average size of 100 nm and a rather narrow size evolution. The resulting nanoparticles do not show acute toxicity to primary cultures of fibroblasts and hepatocytes, epithelial-like cell line L-929 and granulocyte-like culture of HL-60 at a concentration of 109 NPs/mL. SeNPs at a concentration of 1010 NPs/mL reduced the viability of HL-60 cells by no more than 10% and did not affect the viability of the primary culture of mouse granulocytes, and did not have a genotoxic effect on progenitor cells. The addition of SeNPs can affect the production of reactive oxygen species (ROS) by mouse bone marrow granulocytes, modulate the proportion of granulocytes with calcium spikes and enhance fMLF-induced granulocytes degranulation. SeNPs can modulate the effect of IgG on the physiological responses of granulocytes. We studied the expression level of genes associated with inflammation and cell stress. SeNPs increase the expression of catalase, NF-κB, Xrcc5 and some others; antibodies enhance the effect of SeNPs, but IgG without SeNPs decreases the expression level of these genes. This fact can be explained by the interaction between SeNPs and IgG. It has been established that antibodies interact with SeNPs. We showed that antibodies bind to the surface of selenium nanoparticles and are present in aqueous solutions in a bound form from DLS methods, ultraviolet-visible spectroscopy, vibrational-rotational spectrometry, fluorescence spectrometry, and refractometry. At the same time, in a significant part of the antibodies, a partial change in the tertiary and secondary structure is observed. The data obtained will allow a better understanding of the principles of the interaction of immune cells with antibodies and SeNPs and, in the future, may serve to create a new generation of immunomodulators.

The effect of high temperature on kinetics of reactive species generation in patients with type 2 diabetes.

The excessive amount of reactive species under chronic inflammation, which are accompanied by an increase body temperature, lead to diabetic complications. Phagocyte NADPH oxidase is the key enzyme in these processes. The role of high temperature in its regulation in diabetes is not clear. The aim was to investigate the effect of high temperature on NADPH-oxidase-dependent generation of reactive species in diabetic patients. Chemiluminescent method was applied to assess respiratory burst kinetics initiated by opsonized zymosan in blood or phorbol ester in isolated granulocytes. Analyzing ROC curves, the main predictors and changes in stages of activation of NADPH oxidase were determined. Phosphoisoforms of p47phox and p67phox were quantified by immunoblotting. Response to opsonized zymosan was lower in all subjects at 40 °C vs 37 °C, its kinetic parameters (except Tmax) were higher in blood of patients vs controls. Response rate was the main significant predictor to distinguish groups of subjects at 40 °C indicating NADPH oxidase upregulation in diabetes. Ca2+-dependent generation of reactive species by cells increased in both groups at 40 °C vs 37 °C, kinetic parameters were higher in patients. Initial phospho-p47phox level was higher in patient cells vs ones in controls. It was increased by ionomycin, phorbol ester, or 40 °C in control cells and unchanged in patient ones. Phospho-p67phox level was unchangeable in intact cells of healthy donors and patients at both temperatures. Excessive amounts of reactive species in patient cells were the consequence of granulocyte priming due to p47phox phosphorylation. Thus, high temperature decreased phagocytosis- and enhanced Ca2+-dependent generation of reactive species making the differences between controls and patients less pronounced. The effect of temperature on the generation of reactive species in blood granulocytes is associated with activity of NADPH oxidase that can be a prospective therapeutic target for pathologies accompanied by inflammation including type 2 diabetes.

Elucidating the picocyanobacteria salinity divide through ecogenomics of new freshwater isolates.

BACKGROUND: Cyanobacteria are the major prokaryotic primary producers occupying a range of aquatic habitats worldwide that differ in levels of salinity, making them a group of interest to study one of the major unresolved conundrums in aquatic microbiology which is what distinguishes a marine microbe from a freshwater one? We address this question using ecogenomics of a group of picocyanobacteria (cluster 5) that have recently evolved to inhabit geographically disparate salinity niches. Our analysis is made possible by the sequencing of 58 new genomes from freshwater representatives of this group that are presented here, representing a 6-fold increase in the available genomic data. RESULTS: Overall, freshwater strains had larger genomes (≈2.9 Mb) and %GC content (≈64%) compared to brackish (2.69 Mb and 64%) and marine (2.5 Mb and 58.5%) isolates. Genomic novelties/differences across the salinity divide highlighted acidic proteomes and specific salt adaptation pathways in marine isolates (e.g., osmolytes/compatible solutes - glycine betaine/ggp/gpg/gmg clusters and glycerolipids glpK/glpA), while freshwater strains possessed distinct ion/potassium channels, permeases (aquaporin Z), fatty acid desaturases, and more neutral/basic proteomes. Sulfur, nitrogen, phosphorus, carbon (photosynthesis), or stress tolerance metabolism while showing distinct genomic footprints between habitats, e.g., different types of transporters, did not obviously translate into major functionality differences between environments. Brackish microbes show a mixture of marine (salt adaptation pathways) and freshwater features, highlighting their transitional nature. CONCLUSIONS: The plethora of freshwater isolates provided here, in terms of trophic status preference and genetic diversity, exemplifies their ability to colonize ecologically diverse waters across the globe. Moreover, a trend towards larger and more flexible/adaptive genomes in freshwater picocyanobacteria may hint at a wider number of ecological niches in this environment compared to the relatively homogeneous marine system.

α-cyanobacteria possessing form IA RuBisCO globally dominate aquatic habitats.

RuBisCO (ribulose 1,5-bisphosphate carboxylase/oxygenase) is one the most abundant enzymes on Earth. Virtually all food webs depend on its activity to supply fixed carbon. In aerobic environments, RuBisCO struggles to distinguish efficiently between CO2 and O2. To compensate, organisms have evolved convergent solutions to concentrate CO2 around the active site. The genetic engineering of such inorganic carbon concentrating mechanisms (CCMs) into plants could help facilitate future global food security for humankind. In bacteria, the carboxysome represents one such CCM component, of which two independent forms exist: α and ß. Cyanobacteria are important players in the planet's carbon cycle and the vast majority of the phylum possess a ß-carboxysome, including most cyanobacteria used as laboratory models. The exceptions are the exclusively marine Prochlorococcus and Synechococcus that numerically dominate open ocean systems. However, the reason why marine systems favor an α-form is currently unknown. Here, we report the genomes of 58 cyanobacteria, closely related to marine Synechococcus that were isolated from freshwater lakes across the globe. We find all these isolates possess α-carboxysomes accompanied by a form 1A RuBisCO. Moreover, we demonstrate α-cyanobacteria dominate freshwater lakes worldwide. Hence, the paradigm of a separation in carboxysome type across the salinity divide does not hold true, and instead the α-form dominates all aquatic systems. We thus question the relevance of ß-cyanobacteria as models for aquatic systems at large and pose a hypothesis for the reason for the success of the α-form in nature.

α9α10 nicotinic acetylcholine receptors regulate murine bone marrow granulocyte functions.

Polymorphonuclear neutrophilic granulocytes (PMNs) are extremely important in defense of the organism against infections and in inflammatory processes including neuroinflammation and pain sensation. Different subtypes of nicotinic acetylcholine receptors (nAChRs) are involved in modulation of PMN activities. Earlier we determined expression of α2-7, α9, ß3, ß4 subunits and regulatory role of α7 and α3ß2 nAChR subtypes in functions of inflammatory PMNs. Other authors detected mRNA of α9 subunit in bone marrow neutrophils (BM-PMNs). Murine BM-PMNs coming out from the bone marrow, where they develop, to blood were characterized as mature. There was no data for α10 and for the presence of functionally active α9α10 nAChRs in BM-PMNs. Here we detected for the first time mRNA expression of the α10 nAChR subunit in BM-PMNs and confirmed the expression of mRNA for α9 nAChR. With the help of α-conotoxins RgIA and Vc1.1, highly selective antagonists of α9α10 nAChRs, we have revealed participation of α9 and/or α9α10 nAChRs in regulation of cytosolic Ca2+ concentration, cell adhesion, and in generation of reactive oxygen species (ROS). Nicotine, choline, RgIA, and Vc1.1 induced Ca2+ transients in BM-PMNs, enhanced cell adhesiveness and decreased production of ROS indicating involvement of α9, possibly co-assembled with α10, nAChRs in the BM-PMN activity for recruitment and cytotoxicity.

Effect of orthostasis on the regulation of skin blood flow in upper and lower extremities in human.

OBJECTIVE: The research is aimed to investigate interactions between cardiovascular signals and to assess contributions of central and local mechanisms to skin blood flow regulation in upper and lower extremities at rest and under orthostasis. METHODS: Heart rate variability, respiration, forearm, and foot skin blood flow were assessed at rest and during postural test in 25 healthy volunteers. Spectral analysis was performed. Phase synchronization degree of analyzed signals was determined by group phase wavelet coherence function. RESULTS: Skin blood flow was lower on foot at rest and during postural test than on forearm. High-frequency component of heart rate variability was higher at ~0.3 Hz during postural test versus rest. Blood flow oscillation amplitudes on the foot were lower in frequency range including respiratory interval at rest than on forearm. Postural exposure increased amplitude of foot blood flow oscillations in respiratory interval and decreased amplitudes in cardiac interval versus rest. Orthostasis increased group wavelet phase coherence between foot blood flow and heart rate variability or respiration, as well as between forearm and foot blood flow at 0.3 Hz corresponding to respiration. CONCLUSIONS: The contribution of central mechanisms associated with respiration to blood flow regulation increased in lower extremities during orthostasis.

Modified kinetics of generation of reactive species in peripheral blood of patients with type 2 diabetes.

Level of reactive species in blood is an important pathogenic factor in diabetes mellitus leading to dysfunctions of vascular endothelial and smooth muscle cells and coagulation system abnormality. A massive release of reactive species (respiratory burst), catalyzed by NADPH oxidase in blood phagocytes, is not well understood in diabetes. The work aimed to study kinetics of response to microbial particles in blood to specify changes in regulatory mechanisms of generation of reactive species in patients with type 2 diabetes. Production of reactive species in blood and isolated granulocytes was measured by luminol-dependent chemiluminescence. Respiratory burst was initiated by serum opsonized zymosan in blood samples and phorbol ester in cell samples. Kinetic parameters were calculated from experimental kinetic curves of chemiluminescence intensity. ROC curve analysis and mathematical modeling were used to reveal the most significant predictors and clarify specific mechanisms of NADPH oxidase activation. It was shown that kinetic parameters of response to opsonized zymosan (lag-time, response rate, amplitude, production of reactive species) were higher in blood of patients than controls. Amplitude and response rate were the most statistically significant predictors for distinguishing patients and controls at high glucose. It indicated NADPH oxidase activation was the target of hyperglycemia. Mathematical modeling showed hyperglycemia increased stability of NADPH oxidase complex, decreased synchronization of its assembling and elevated neutrophil capacity to phagocytosis in patients. Weak or no dependence of response kinetics on ionomycin concentration was shown in patients indicating changed Ca2+-dependent mechanism of NADPH oxidase activation. Hyperglycemia in type 2 diabetes causes disturbances in mechanisms of NADPH oxidase activation associated with both phagocytosis and the state of intracellular signaling systems, including Ca2+-dependent. We suggest that NADPH oxidase in blood granulocytes can be a promising target for clinical intervention improving management of diabetic complications associated with inflammation.

Metagenomic Analysis of Virioplankton from the Pelagic Zone of Lake Baikal.

This study describes two viral communities from the world's oldest lake, Lake Baikal. For the analysis, we chose under-ice and late spring periods of the year as the most productive for Lake Baikal. These periods show the maximum seasonal biomass of phytoplankton and bacterioplankton, which are targets for viruses, including bacteriophages. At that time, the main group of viruses were tailed bacteriophages of the order Caudovirales that belong to the families Myoviridae, Siphoviridae and Podoviridae. Annotation of functional genes revealed that during the under-ice period, the "Phages, Prophages, Transposable Elements and Plasmids" (27.4%) category represented the bulk of the virome. In the late spring period, it comprised 9.6% of the virome. We assembled contigs by two methods: Separately assembled in each virome or cross-assembled. A comparative analysis of the Baikal viromes with other aquatic environments indicated a distribution pattern by soil, marine and freshwater groups. Viromes of lakes Baikal, Michigan, Erie and Ontario form the joint World's Largest Lakes clade.

Delayed kinetics of phagocytosis related respiratory burst in blood is a distinctive feature of moderate exacerbation of bronchial asthma.

Atopic bronchial asthma based on allergy history and chronic inflammation is hazardous to patients due to the risk of exacerbation. The sign of severe exacerbation is considered an abundant number and high activity of granulocytes in respiratory system and blood. Relationships between the ability of cells in blood to produce reactive radicals and their metabolites and the severity of asthma remain largely unclear. Kinetics of respiratory burst evoked by microbe particles in blood samples of patients was studied to reveal the most significant predictors distinguishing states of moderate exacerbation and out of exacerbation. Asthmatic patients with exacerbation (n = 18) or out of exacerbation (n = 62) and healthy individuals (n = 43) were characterized on respiratory function, cell count in blood and kinetics of generation of reactive radicals and their metabolites during phagocytosis. Mean values of respiratory parameters forced expiratory volume in 1 s and peak expiratory flow rate in patients with exacerbation were significantly differed compared with same of patients out of exacerbation and healthy individuals. Mean values of cell count in blood did not significantly differed in patients with exacerbation and out of exacerbation. Receiver operating characteristic analysis showed that both cell count and respiratory indexes did not discriminate patients with exacerbation from out of exacerbation. A delayed response to opsonized zymosan was revealed in patients with exacerbation compared to other examinees: lengthened lag-time and Tmax, reduced production of reactive species. Tmax was the most statistically significant predictor to discriminate bronchial asthma exacerbation from bronchial asthma out of exacerbation (area under curve >90%, p < 10-5) and controls (area under curve >80%, p < 10-5). Thus kinetic parameters of the phagocyte response to opsonized zymosan in the whole blood are the best predictors of bronchial asthma exacerbation in comparison with respiratory parameters and blood cell count. This test can be used for immunological monitoring of bronchial asthma status to prevent exacerbation.

Accelerated reactivity of blood granulocytes in patients with atopic bronchial asthma out of exacerbation.

Reactive oxygen species (ROS) are important in bronchial asthma (BA) pathogenesis owing to accumulation of activated granulocytes in the lungs. But the ROS-producing activity of the cells is insufficiently understood in the blood of BA patients. This study analyzes the kinetics of phagocyte respiratory burst in the blood to improve the methods of BA patients monitoring. Patients with atopic BA out of exacerbation (n=60) and healthy controls (n=43) were recruited. The time-course of respiratory response to opsonized zymosan (OZ) was recorded in the whole blood using luminol-dependent chemiluminescence (CL), and its activation kinetics (lag-time, rate, amplitude, ROS production) was calculated. The discriminative power of ROS generation kinetics was defined by Receiver Operating Characteristic (ROC) curve analysis. Standard physiological respiratory parameters of patients did not differ from the controls. More rapid response to OZ was recorded in BA patient samples versus the controls. The primed state of phagocytes in the blood of BA patients was corroborated by significant weakening formyl peptide priming effect. The adhesion of granulocytes to cultured human endothelial cells was two-fold higher in BA patients versus controls. ROC curve analysis exhibited good discriminative effectiveness of the CL kinetics to compare BA individuals with the controls. The highest power (86% sensitivity and 90% specificity) was achieved at a linear combination of the parameters. We assume that the assessment of phagocyte reactivity based on the analysis of the response kinetic profile is a good test for monitoring of the state in BA patients.

Vascular Pathology of Ischemia/Reperfusion Injury of Rat Small Intestine.

Ischemia/reperfusion (I/R) injury of the small intestine caused by occlusion of the superior mesenteric artery affects the intestinal tissue as well as components of the blood circulatory system from the microvasculature to mesenteric vessels. The aim of this work was to study the correlation between the dynamics of destruction development in the intestinal tissue, microvasculature, and mesenteric vessels in I/R of the small intestine. The microvasculature was analyzed by whole-organ continuous monitoring of the intestinal mucosal blood perfusion by laser Doppler flowmetry during the entire I/R. Real-time RT-PCR was used to assess gene expression of NF-κB, caspase-3, Ki67, and TNF-α in blood vessels. At the start of reperfusion, the first targets to be disrupted are microvessels in the apical villi. Injury of the apical part of the microcirculatory bloodstream correlates with the reduction in intestinal mucosal blood perfusion, which occurred simultaneously with apical villous destruction. By the end of the reperfusion period, the low intestinal mucosal blood perfusion is mirrored by the destruction of the microvasculature and mucosal structures in the entire organ. The development of mesenteric vessel injury is characterized by a change in NO metabolism and damaged endothelial cells concomitant with an alteration in the expression of genes encoding NF-κB, caspase-3, and Ki67 by the end of the reperfusion period. In I/R injury, detrimental effects on the intestinal tissue, microvasculature, and mesenteric vessels develop and exhibit common mechanisms of function, which show strong correlations.

First detection of benthic cyanobacteria in Lake Baikal producing paralytic shellfish toxins.

Cyanobacteria were screened from the surface of diseased sponges, stone and bedrock in Lake Baikal for the presence of saxitoxin using enzyme-linked immunosorbent assay. In sequel, eight paralytic shellfish toxin (PST) variants were identified using a MALDI mass spectrometry. Microscopic examination found that Tolypothrix distorta dominated in the biofouling samples. PCR and sequencing detected sxtA gene involved in saxitoxin biosynthesis, thereby providing evidence of the PST producing potential of Baikal cyanobacterial communities inhabiting different substrates.

Oscillations of Skin Microvascular Blood Flow in Patients with Asthma.

OBJECTIVE: This research is aimed at studying the features of skin blood flow oscillations in patients with severe persistent atopic BA during a period of fine control over symptoms. METHODS: The study of microcirculation was carried out by LDF at rest and in response to a transient ischemia in 20 patients. The time-amplitude adaptive wavelet analysis of the blood flow oscillations was conducted to elucidate the peculiarities of microcirculatory regulation system functioning. RESULTS: No significant changes were revealed for SBP and the oscillation amplitudes in the cardiac (0.6-2 Hz) and respiratory (0.145-0.6 Hz) intervals, both at rest and in response to transient ischemia, in patients compared to the control group. A consistent twofold decrease in the oscillation amplitudes was found in the neurogenic (0.021-0.052 Hz) interval at rest, as well as in the myogenic (0.052-0.145 Hz) and NO-dependent endothelial (0.0095-0.021 Hz) intervals both at rest and during the postocclusive reactive hyperemia in patients with lung obstruction (FEV1 < 80%) in comparison with a control group. CONCLUSIONS: The amplitudes of skin blood flow oscillations in the myogenic, neurogenic and NO-dependent endothelial intervals in patients with obstruction are different from those in patients without obstruction.

Diversity of cyanobacterial species and phylotypes in biofilms from the littoral zone of Lake Baikal.

The majority of naturally occurring biofilms contain numerous microorganisms that have not yet been cultured. Additionally, there is little information available regarding the genetic structure and species diversity of these communities. Therefore, we characterised the species diversity, structure and metagenome of biofilms grown on stones and steel plates in the littoral zone of Lake Baikal (East Siberia, Russia) by applying three different approaches. First, light microscopy enabled identification of the species diversity of biofilm-forming cyanobacteria on different substrates with the dominance of Rivularia rufescens, Tolypothrix limbata, Chamaesiphon fuscus, Ch. subglobosus, and Heteroleibleinia pusilla. Additionally, scanning electron microscopy was used to show the spatial structure of biofilms. Finally, sequence analysis of 30,660 16S rRNA clones indicated a high diversity within the biofilm communities, with the majority of the microbes being closely related to Cyanobacteria (8-46% sequences), Proteobacteria (14-43%), and Bacteroidetes (10-41%). Rivularia sp., Pseudanabaena sp., and Chamaesiphon spp. were the dominant cyanobacterial phylotypes.

Age-related changes of skin blood flow during postocclusive reactive hyperemia in human.

BACKGROUND/PURPOSE: The objective was to study age-related alterations in the time-amplitude characteristics of the oscillatory components of peripheral blood flow in healthy humans during postocclusive reactive hyperemia. METHODS: Forearm blood skin perfusion was studied by the method of laser Doppler flowmetry in 120 healthy volunteers, who were divided into two age groups: young (19-30 years old; n = 82) and middle-age (30-60 years old; n = 38). The forearm occlusion approach was used to reveal the reaction of the microvascular bed to transient ischemia. To estimate the age-related changes of separate oscillatory components of LDF signals during postocclusive reactive hyperemia, we applied adaptive time-amplitude wavelet analysis. RESULTS: A statistically significant increase in the skin blood perfusion after occlusion removal was revealed: 4-fold in the 1st and 3.5-fold in the 2nd group respectively. Both the amplitude of blood flow oscillations at rest and the postocclusive growth of the amplitude showed an age-related decline in the frequency intervals associated with the myogenic (0.052-0.145 Hz), sympathetic (0.021-0.052 Hz) and endothelial (0.0095-0.021 Hz) activity. CONCLUSION: The time-amplitude characteristics of the oscillatory components of peripheral blood flow depend on age, with the dependence becoming more evident after a transient ischemia.

Stratified distribution of nutrients and extremophile biota within freshwater ice covering the surface of Lake Baikal.

Biological entities and gradients of selected chemicals within the seemingly barren ice layers covering Lake Baikal were investigated. Ice cores 40-68 cm long were obtained from in shore and offshore sites of Southern Lake Baikal during the cold period of a year (March-April) in 2007 and 2008. In microscopic observations of the melted ice, both algae and bacteria were found in considerable numbers (>10(3) cells/L and >10(4) cells/ml, respectively). Among all organisms found, diatom was generally the most predominant taxon in the ice. Interestingly, both planktonic and benthic algae were present in considerable numbers (2-4×10(4) cells/L). Dominant phototrophic picoplankton were comprised of small green algae of various taxa and cyanobacteria of Synechococcus and Cyanobium. The bacterial community consisted mostly of short rod and cocci cells, either free-living or aggregated. Large numbers of yeast-like cells and actinomycete mycelium were also observed. Concentrations of silica, phosphorus, and nitrate were low by an order of magnitude where biota was abundant. The profile of the ice could be interpreted as vertical stratification of nutrients and biomass due to biological activities. Therefore, the organisms in the ice were regarded to maintain high activity while thriving under freezing conditions. Based on the results, it was concluded that the freshwater ice covering the surface of Lake Baikal is considerably populated by extremophilic microorganisms that actively metabolize and form a detritus food chain in the unique large freshwater ecosystem of Lake Baikal.

Ecological development and genetic diversity of Microcystis aeruginosa from artificial reservoir in Russia.

Microcystis aeruginosa is a well-known Cyanobacterium responsible for the formation of toxic water blooms around the world. Shallow, warm, and eutrophic reservoirs provide the most favourable conditions for M. aeruginosa development. Numerous studies have been devoted to this species, but there still is a necessity to develop additional approaches for the monitoring of cyanobacteria in reservoirs. In this study, M. aeruginosa in the water column of a hypereutrophic Siberian reservoir was investigated by fluorescence, light, and electron microscopy as well as genetic analysis using a mcyE marker. Here, we demonstrate the genetic diversity and features of the fluorescence spectra for different ecotypes of this species. We suggest that a fluorescence approach can be used to identify M. aeruginosa in a natural environment in order to increase the effectiveness of ecological monitoring and water quality evaluation.

Time-amplitude analysis of skin blood flow oscillations during the post-occlusive reactive hyperemia in human.

The perfusion of forearm skin with blood was studied by the method of laser Doppler flowmetry (LDF) in 94 healthy volunteers. We studied the reaction of the microvascular bed to the transient ischemia, which was initiated by the forearm occlusion. After occlusion, we registered, on average, a 4-fold increase of skin blood perfusion as compared to the level of this parameter at rest. In the study, we also analyzed changes of the oscillatory components of LDF signals during post-occlusive reactive hyperemia; these components were revealed with the adaptive time-amplitude wavelet analysis. It was found that the time needed for oscillations to reach their maximal amplitude is different for each of the frequency intervals examined. After occlusion, a statistically significant rise of the amplitude of blood flow oscillations was revealed-for the frequency intervals corresponding to the cardiac (0.6-2 Hz), respiratory (0.145-0.6 Hz), myogenic (0.052-0.145 Hz), sympathetic (0.021-0.052 Hz), and endothelial (0.0095-0.021 Hz) activity (a more than 11-, 8-, 6-, 3-, and 6-fold increase, respectively, as compared to the state of rest). The method applied here for the analysis of oscillatory components of LDF signals can, therefore, be used to study the dynamics of oscillations of peripheral blood flow under various functional tests.