[Genome Stability of Bacillus velezensis after Two-Year Exposure in Open Space].

Spore-forming bacteria have a unique resistance to negative environmental conditions, including aggressive space factors, and are an excellent model for studying adaptation mechanisms and survival strategies at the molecular level. The study analyzed the genome of Bacillus velezensis, which remained viable after a 2-year exposure in outer space on the outer surface of the ISS as part of the Test space experiment. A comparative analysis of the draft genomes of the exhibit strain and the ground control did not reveal significant changes; the average nucleotide identity was 99.98%, which indicates the ability of microorganisms to maintain genome stability in space conditions, due to both increased stress resistance of bacterial spores and efficient operation of the system of repair of accumulated changes. The study of a single nucleotide polymorphism in the genome of B. velezensis revealed nine point substitutions, three of which are in intergenic regions, six in protein-coding genes, three of them are missense mutations, two nucleotide deletions leading to a shift in the reading frame, and one synonymous substitution. The profiles of the housekeeping genes were determined during MLST typing and it was found that the allelic profiles obtained for B. velezensis T15.2 and 924 strains do not correspond to any of the previously described sequence types. The presented results indicate the ability of B. velezensis bacteria to maintain the viability of spores and the integrity of the genome for a long time under extreme conditions of outer space, which is important for the problem of planetary protection, as well as the potential possibility of performing biotechnological processes based on B. velezensis during space exploration.

Symmetric Dimethylarginine Assay Validation, Stability, and Evaluation as a Marker for the Early Detection of Chronic Kidney Disease in Dogs.

BACKGROUND: Symmetric dimethylarginine (SDMA) is a small molecule formed by methylation of arginine, and released into blood during protein degradation. SDMA is primarily eliminated by renal excretion and is a promising endogenous marker of glomerular filtration rate (GFR). OBJECTIVES: To validate an assay for SDMA measurement, determine stability of SDMA in blood, and compare SDMA with serum creatinine concentration (sCr) and GFR for early detection of decreasing kidney function in dogs with chronic kidney disease (CKD). ANIMALS: Eight male dogs affected with X-linked hereditary nephropathy and 4 unaffected male littermates. METHODS: Prospective study validating SDMA measurement using liquid chromatography-mass spectrometry, assessing stability of SDMA in serum and plasma, and serially determining sCr, SDMA, and GFR (using iohexol clearance) in dogs during progression from preclinical disease to end-stage renal failure. Correlations were determined using linear regression. Timepoints at which sCr, SDMA, and GFR identified decreased renal function were compared using defined cutoffs, trending in an individual dog, and comparison with unaffected littermates. RESULTS: Symmetric dimethylarginine was highly stable in serum and plasma, and the assay demonstrated excellent analytical performance. In unaffected dogs, SDMA remained unchanged whereas in affected dogs, SDMA increased during disease progression, correlating strongly with an increase in sCr (r = 0.95) and decrease in GFR (r = -0.95). Although trending improved sCr's sensitivity, SDMA identified, on average, <20% decrease in GFR, which was earlier than sCr using any comparison method. CONCLUSIONS AND CLINICAL IMPORTANCE: Symmetric dimethylarginine is useful for both early identification and monitoring of decreased renal function in dogs with CKD.

Mortality risk in adults with newly diagnosed and chronic epilepsy: a population-based study.

BACKGROUND: We analyzed mortality in adult patients with newly diagnosed and chronic epilepsy over a 13-year period. METHODS: Eighty-one patients aged ≥ 20 years with newly diagnosed epilepsy and 309 adult patients with chronic epilepsy were originally identified from population-based incidence and prevalence studies conducted in Tartu between 1994 and 1996. Patients with epilepsy were followed until the date of death or until the end of 2007. The standardized mortality ratio (SMR) was analyzed for both cohorts. The influences of age at diagnosis, sex, epilepsy syndrome, seizure type, risk factors and treatment compliance on the SMR were also investigated. RESULTS: The SMR was significantly increased in both cohorts, but was higher in patients with chronic epilepsy (SMR 3.1; 95% confidence interval [CI] 2.5-3.8) relative to patients with newly diagnosed epilepsy (SMR 2.6; 95% CI 1.8-3.5). In the newly diagnosed epilepsy cohort, the increased mortality risk was more pronounced in patients with complex partial seizures (SMR 5.6; 95% CI 2.4-11.0). In the chronic epilepsy cohort, the mortality risk was higher in patients with secondary generalized tonic-clonic seizures (SMR 3.4; 95% CI 2.5-4.5). Non-compliant patients had twice the mortality risk (SMR 4.2; CI 95% 2.7-6.2) compared to those who were on anticonvulsant treatment. CONCLUSIONS: Mortality rates are higher in people with newly diagnosed and chronic epilepsy. Mortality risks should be discussed with patients with epilepsy, especially if anticonvulsant treatment is refused despite recurrent seizures.

[Intragastric pH analysis during alkaline test].

A new technique for intragastric pH-gram analysis during alkaline test is described. The technique is based on a mathematical model of the acid-alkaline balance in the body of stomach. The model provides intragastric pH-gram approximation by the least-squares method. This makes it possible to determine the specific rate of acid production in the stomach and accurately calculate the alkaline time. The structure of a hardware-software system for implementation of the new technique is developed. The suggested algorithms can be implemented on the basis of the Gastroscan-5M acidogastrometer.

17beta-Oestradiol stimulation of G-proteins in aged and Alzheimer's human brain: comparison with phytoestrogens.

The neuroprotective action of oestrogens and oestrogen-like compounds is in the focus of basic and clinical research. Although such action has been shown to be associated with neuronal plasma membranes, the implication of G-proteins remains to be elucidated. This study revealed that micromolar concentrations (microM) of 17beta-oestradiol and phytoestrogens, genistein and daidzein, significantly (P < 0.05) stimulate G-proteins ([(35)S]GTP gamma S binding) in the post-mortem hippocampal membranes of age-matched control women with the respective maximum effects of 28, 20 and 15% at 10 microM. In the frontocortical membranes, the stimulation of G-proteins did not differ significantly from that in hippocampal membranes. Although in the hippocampus and frontal cortex of the Alzheimer's disease (AD) women's brain, 10 microM 17beta-oestradiol produced significantly (P < 0.05) lower stimulation of G-proteins than in the control regions, stimulation by phytoestrogens revealed no remarkable decline. 17beta-Oestradiol, genistein and daidzein revealed a selective effect on various G-proteins (G(alphas), G(alpha o), G(alpha i1) or G(alpha 11) plus G(beta 1 gamma 2)) expressed in Sf9 cells. At a concentration of 10 microM, 17beta-oestradiol suppressed the H(2)O(2) and homocysteine stimulated G-proteins in the frontocortical membranes of control women to a greater extent than phytoestrogens. In AD, the suppressing effect of each compound was lower than in the controls. In the cell-free systems, micromolar concentrations of phytoestrogens scavenged OH(*) and the 2.2-diphenyl-1-picrylhydrazyl free radical (DPPH(*)) more than 17beta-oestradiol did. In the frontocortical membranes of control women, the 20 microM 17beta-oestradiol stimulated adenylate cyclase with 20% maximal effect, whereas, in AD, the effect was insignificant. Genistein did not stimulate enzyme either in control or AD frontocortical membranes. Our data confirm that the agents stimulate G-proteins in control and AD women's brains, although 17beta-oestradiol and phytoestrogens have similarities and differences in this respect. We suggest that, besides the ER-dependent one, the ER-independent antioxidant mechanism is responsible for the oestrogen stimulation of G-proteins in the brain membranes. Both of these mechanisms could be involved in the neuroprotective signalling of oestrogens that contributes to their preventive/therapeutic action against postmenopausal neurological disorders.

[Intragastric pH probe functioning in studies of solutions with nonuniform pH].

The goal of this work was to study the antimony electrode functioning in solution with nonuniform pH. Two research methods were developed. The first method was based on the use of the current-voltage curve of the pH probe. This method made it possible to study the dependence of the pH probe functioning on the contact patch size. The second method was based on experimental simulation of the pH probe contact with solutions with different pH. This method made it possible to study the dependence of the pH probe functioning on the gastric juice concentration. It was found that changes in sodium bicarbonate concentration had greater effect on the electrode functioning than changes in the gastric juice concentration.

Metallic conduction through engineered DNA: DNA nanoelectronic building blocks.

A novel way of engineering DNA molecules involves substituting the imino proton of each base pair with a metal ion to obtain M-DNA with altered electronic properties. We report the first direct evidence of metalliclike conduction through 15 microm long M-DNA. In contrast, measurements on B-DNA give evidence of semiconducting behavior with a few hundred meV band gap at room temperature. The drastic change of M-DNA conductivity points to a new degree of freedom in the development of future molecular electronics utilizing DNA, such as creating all-DNA junction devices for use as nanoelectronic building blocks.

Electronic transport in Y-junction carbon nanotubes

Electronic transport measurements were performed on Y-junction carbon nanotubes. These novel junctions contain a large diameter tube branched into smaller ones. Independent measurements using good quality contacts on both individual Y junctions and many in parallel show intrinsic nonlinear transport and reproducible rectifying behavior at room temperature. The results were modeled using classic interface physics for a junction with an abrupt change in band gap due to the change in tube diameter. These Y-junction tubes represent new heterojunctions for nanoelectronics.

Serotonin-induced short- and long-term sensitization in the crab Chasmagnathus.

On sudden presentation of a passing shadow (a visual danger stimulus), the crab Chasmagnathus granulatus responds with a running reaction that we term an escape response (ER). A single administration of 5-HT in a dose range between 0.005 and 0.1 microgram/g enhances the ER at 30 min, and a dose of 10-15 micrograms/g has a similar effect for at least 24 h. The classical 5-HT antagonist cyproheptadine (CYP) given within a dose range 0.01-2.0 micrograms/g has no effect on ER at 30 min, but 0.5 microgram/g blocks the 5-HT-induced short-term sensitization. An enhancing effect of CYP on the ER is shown at 24 h with 1.0-2.0 micrograms/g doses. An explanation of this results in terms of a model similar to that proposed for Aplysia, that is two sensitizing processes linked in parallel and mediated by two different types of 5-HT receptors is discussed. Results indicate that the sensitizing effect of 5-HT is confined to high responders, thus suggesting that crabs of a same population have different degrees of sensitivity to the drug according to their different degrees of reactivity to the visual danger stimulus.

Habituation and sensitization to an electrical shock in the crab Chasmagnathus. Effect of background illumination.

The escape response of the crab Chasmagnathus granulatus, elicited by an electrical leg-shock, wanes as a consequence of repeated stimulation, and the decrease persists after a 24-h rest interval. Results concerning stimulus specificity in within- and between-sessions habituation strongly indicate that neither motor fatigue nor sensory adaptation nor damage can account for the response waning, which thus meets the major criteria of habituation. A comparison between the escape response habituation to leg-shock and that to a shadow passing overhead discloses clearcut differences. The shock curve shows an initial hump positively related to stimulus intensity, suggesting that a shock-induced sensitization along with habituation subserves the response curve. The curve asymptote is remarkably high and, unlike the initial waning portion, not greatly affected by treatment changes, hinting that the response may be the combination of an escape response and a basal one (an undirected burst of activity). The waning portion is higher when shocks are given in dark than when given in light, but this effect is not considered as an extra eliciting-stimulus-induced sensitization, since the reactivity enhancement depends necessarily on the darkness-shock concomitance, regardless of prior exposure to darkness. Accordingly, the hypothesis that crab habituates not only to a shock but to a complex stimulus (shock + background illumination) is put forward and discussed.

Morphine and GABA: effects on perception, escape response and long-term habituation to a danger stimulus in the crab Chasmagnathus.

Prior results (37) showed that morphine pretreatment reduces the escape response of the crab Chasmagnathus to a shadow passing overhead and prevents the acquisition of a long-term habituation. These results were explained by a reduction in the danger signalled by the stimulus, and to test this hypothesis methods other than morphine injection were used herein to abolish response during training. GABA pretreatment induced a dose-dependent reduction in responsiveness to the danger stimulus, and instances of autotomy were shown with doses larger than 12 micrograms/g. A response was rarely displayed with a 9 micrograms GABA/g dose given 5 min before training, but long-term memory was acquired. In one experiment, both morphine and GABA pretreatment produced similar mild response inhibition during training, but morphine, not GABA impaired long-term habituation. Morphine administered immediately after training had no amnesic effect. These results support the hypothesis that morphine effects may be explained by transient disruption between the stimulus and its danger meaning, ruling out alternative explanations such as response inhibition or amnesia due to either storage or retrieval failure.