[Regulation of sulfates, hydrogen sulfide and heavy metals in technogenic reservoirs by sulfate-reducing bacteria].

Sulfate-reducing bacteria Desulfovibrio desulfuricans Ya-11 in the presence of sulfates and organic compounds in the medium reduce sulfates to hydrogen sulfide (dissimilatory sulfate reduction). Heavy metals in concentration over 2 mM inhibit this process. Pb2+, Zn2+, Ni2+, Co2+, Fe2+ and Cd2+ ions in concentration 1-1.5 mM display insignificant inhibiting effect on sulfate reduction process, and metals precipitate in the form of sulfides. At concentrations of heavy metals 2-3 mM one can observe a decrease of sulfates reduction intensity, and a percent of metals binding does not exceed 72%. Obtained results give reason to confirm, that sulfate-reducing bacteria play an important role in regulation of the level of sulfates, hydrogen sulfide and heavy metals in reservoirs and they may be used for purification of water environment from these compounds.

[Correction of the influence of clinostatic hypokinesis on encephalitic circulatory dynamics during the sleep in patients with hypertensive disease].

During the sleep person is in the state of antiorthostasis, this state provokes vascular cerebral abnormalities (night cerebral hypervolemia). The nature of encephalon blood supply is changing in this horizontal state, in the result there is lowering of hydrostatic blood pressure, rise of the encephalon blood supply and more difficult venous outflow. 60 patients with essential hypertension (1-2 phases) at the age of 64.2 +/- 1.8 years which were in the state of slightly raised upper segment of the head and body during night sleep were examined. By the method of the cerebral venous hemocirculation defined. Conclusion. Cerebral arterial blood flow of examined patients has increased by 25-30% during the sleep, the changes of arterial and cerebral venous hemocirculation have attended hypervolemia. Being in the state of upper (by 10-15 degrees) head of the bed decreases blood redistribution and decreases the extension of cerebral blood circulation's acute abnormalities.

[Regulation of hydrogen sulfide level by acidophobic bacteria of Thiobacillus genus in technogenic reservoirs of sulfur mining regions].

An increase of acidophobic thione bacteria quantity in Rozdil and Yavoriv reservoirs of sulfur mining regions during 2005-2009 years, which correlates with a decrease of hydrogen sulfide content in water surface layers, was shown. The ability of acidophobic bacteria of Thiobacillus genus, isolated from "Yavorivske" lake, to oxidize effectively hydrogen sulfide added into Beijerinck medium instead of thiosulfate, was discovered. It was established, that hydrogen sulfide oxidizing efficiency by Thiobacillus sp. Yav-8, Yav-11 and Yav-14 strains is the highest (78.48-84.56%) when its content in cultivation medium was increased twice: to 2584 mg/l. An increase of sulfur quantity in sodium sulfide form from to six times as compared with its standard content in sodium thiosulfate form in the Beijerinck medium does not lead to the increase of hydrogen sulfide oxidizing efficiency by cells.

[Ethanol into acetaldehyde bioconversion by mutant strains of Hansenula polymorpha Felcao de Morais & Dália Maia].

The influence of some factors (tris-HCl buffer, pH 8.0, concentration, use of different binding agents aeration modes, genetically determined peroxisome degradation damage) on biotransformation efficiency of 0.217 M (1%) ethanol to acetaldehyde at 30 degrees C by Hansenula polymorpha 7-4A (gcrl EAO) strain cells with glucose repression block was investigated. Optimal cultivation conditions for cells were selected. Bioconversion efficiency using 1 M tris-HCl buffer, pH 8.0, was found the highest one as compared with using the buffer in concentrations from 0.1 M to 3 M. The process efficiency when using tris(hydroxymethyl)aminomethane as binding acetaldehyde agent proved much higher than when using sodium bisulfite both at aeration by air stream and incubation on shaker. Using 146 and 179 mutants cells for bioconversion with defects in alcohol oxidase inactivation during macropexophagy stimulated efficiency increase by 5.58% and 8.10%, respectively, as compared with the use of parental 7-4A strain cells.

[Sulfur-oxidizing bacteria of Yavoriv sulfur deposit reservoirs].

Eleven pure bacteria cultures, able to oxidize thiosulfate during growth at pH 7.0-9.4, were isolated from surface layers of Yavoriv sulfur deposit open pit waters. Two cultures proved to be obligate aerobes, but nine cultures performed anaerobic respiration using nitrite, N2O (5 cultures) or only nitrite (4 cultures) as terminal electron acceptors. The growth of all cultures at 22 and 28 degrees C and growth absence at 35, 42 and 55 degrees C was established. All the bacteria are obligate chemolithoauthotrophs, because after cultivation with thiosulfate in the presence ofbiotin, yeast extract, formiate, succinate, arabinose, glucose, fructose and sucrose no growth stimulation was observed, heterothropic growth of any culture was not shown. As to their morphology the cells were bacillary, cytoplasmic membrane was surrounded by three-layer cell wall typical of gram negative bacteria intracellular inclusions, nucleoid, ribosomes and polysomes were also available. On the basis of obtained physiological and morphological characteristics the isolated bacteria cultures were referred to a group of neutrophiles, representatives of genus Thiobacillus, in particular to obligate chemolithoauthotrophs.

[Community-acquired pneumonia in the elderly: the peculiarities of the clinical picture, diagnosis and treatment].

The article presents the results of studying the peculiarities of the clinical picture, diagnosis and treatment for community-acquired pneumonia in the elderly in view of the interaction of combined pathology of respiratory, cardiovascular and urological systems in order to optimize medical-and-diagnostic support of the above-mentioned category of patients in conditions of the pulmonology department of versatile educational establishment. Optimization of all-round inspection of the sick with pneumonia in the advanced age at an initial stage of treatment in conditions of the pulmonology department so as to reveal the aggravation of internal bodies' combined diseases will allow making early correction of aetiotropic, pathogenetic and symptomatic combined pathology therapy available, which reduces the terms of staying at hospital without damage of diagnosis and treatment.

[Biodegradation of oil hydrocarbons by Candida yeast]. / Biodegradatsiia vuhlevodniv nafty drizhdzhamy rodu Candida.

Capability of 14 yeast species to utilize oil hydrocarbons has been analyzed. All strains utilized oil hydrocarbons as a single carbon source. Four strains-destructors that are characterized by higher growth in the presence oil in cultivation medium have been chosen among them. Peroxisomes participation in utilization of oil hydrocarbons by strains-destructors has been shown. Availability of peroxisome key enzymes are characteristic of these strains grown in cultivation medium with oil. Numerous peroxisomes available in the cells of some strains grown in oil cultivation medium have been demonstrated. Utilization of a wide spectrum of oil hydrocarbons has been revealed in all four strains. Two strains are promising to be used for environment purification from oil pollution.

Efficient bioconversion of ethanol to acetaldehyde using a novel mutant strain of the methylotrophic yeast Hansenula polymorpha.

We report the isolation of mutant strains of the methylotrophic yeast Hansenula polymorpha that are able to efficiently oxidize ethanol to acetaldehyde in an intact cell system. The oxidation reaction is catalyzed by alcohol oxidase (AOX), a key enzyme in the methanol metabolic pathway that is typically present only in H. polymorpha cells growing on methanol. At least three mutations were introduced in the strains. Two of the mutations resulted in high levels of AOX in glucose-grown cells of the yeast. The third mutation introduced a defect in the cell's normal ability to degrade AOX in response to ethanol, and thus stabilizing the enzyme in the presence of this substrate. Using these strains, conditions for bioconversion of ethanol to acetaldehyde were examined. In addition to pH and buffer concentration, we found that the yield of acetaldehyde was improved by the addition of the proteinase inhibitor phenylmethylsulfonyl fluoride (PMSF) and by permeabilization of the cells with digitonin. Under optimal shake-flask conditions using one of the H. polymorpha mutant strains, conversion of ethanol to acetaldehyde was nearly quantitative.

Microbial O2- and H2O2-electrode sensors for alcohol assays based on the use of permeabilized mutant yeast cells as the sensitive bioelements.

Two types of alcohol-specific microbial/electrochemical biosensors have been developed using specially constructed mutant cells of the methylotrophic yeast Hansenula polymorpha. The cells were immobilized in a calcium alginate gel, and placed between two membranes on the surface of oxygen or hydrogen peroxide-electrodes. The O2 electrode based biosensor contained mutant cells with strongly elevated alcohol oxidase activity. The peroxide electrode based biosensor consisted of catalase-defective mutant cells which produce hydrogen peroxide in the presence of alcohol. Both types of mutant cells were used in permeabilized form in order to release some components of the cellular respiration system, thus increasing the selectivity of the cellular respiration response to alcohol (cell/O2-biosensor) Permeabilization also increased sensitivity of the signal and shortened the response time (cell/H2O2-biosensor). Cell/O2 biosensors were linear up to 1.2 mM for ethanol and 0.35 mM for methanol, cell/H2O2 biosensors were linear up to 4.0 mM for ethanol, and 1.2 mM for methanol. Results were reproducible, sample pretreatment was not required, and the sensors exhibited good operational and storage stability. The use of sucrose, dulcitol or inositol during the preparation of the sensors resulted in increased stability of cells during their liophilization and storage in the dried state. Both biosensors had similar selectivity towards alcohols in the order of methanol (100%), ethanol (21%), and formaldehyde (12%). No signal was observed with glucose or glycerol as substrates.

Impairment of peroxisome degradation in Pichia methanolica mutants defective in acetyl-CoA synthetase or isocitrate lyase.

Single recessive mutations of the methylotrophic yeast Pichia methanolica acs1, acs2, acs3 and icl1 affecting acetyl-CoA synthetase and isocitrate lyase, and growth on ethanol as sole carbon and energy source, caused a defect in autophagic peroxisome degradation during exposure of methanol-grown cells to ethanol. As a control, a mutation in mdd1, which resulted in a defect of the 'malic' enzyme and also prevented ethanol utilization, did not prevent peroxisome degradation. Peroxisome degradation in glucose medium was unimpaired in all strains tested. Addition of ethanol to methanol-grown cells of acs1, acs2, acs3 and icl1 mutants led to an increase in average vacuole size. Thickening of peroxisomal membranes and tight contacts between groups of peroxisomes and vacuoles were rarely observed. These processes proceeded much more slowly than in wild-type or mdd1 mutant cells incubated under similar conditions. No peroxisomal remnants were observed inside vacuoles in the cells of acs1, acs2, acs3 and icl1 mutants after prolonged cultivation in ethanol medium. We hypothesize that the acs and icl mutants are defective in synthesis of the true effector--presumably glyoxylate--of peroxisome degradation in ethanol medium. Lack of the effector suspends peroxisome degradation at an early stage, namely signal transduction or peroxisome/vacuole recognition. Finally, these defects in peroxisome degradation resulted in mutant cells retaining high levels of alcohol oxidase which further led to increased levels of acetaldehyde accumulation upon incubation of mutant cells with ethanol.