Electrochemical Characteristics of the Oxidation of Sulfur- and Iron-Containing Compounds by Acidophilic Microorganisms.

The electrochemical features of the interactions of sulfur- and iron-containing compounds (ferrous sulfate, elemental sulfur, pyrite tailings, cysteine, sodium thiosulfate) with a model acidophilic consortium, including the genera Leptospirillum, Sulfobacillus, Acidithiobacillus, Ferroplasma, and Acidiplasma, were studied. The method of cyclic voltammetry recorded redox processes at the electrode/solution interface in the presence of the studied sulfur- and iron-containing compounds. In general, the modeling consortium led to the intensification of these processes. The characteristics of the diffuse layer near the electrode/solution interface were studied using electrochemical impedance spectroscopy. The introduction of microorganisms and/or substrates into the supporting electrolyte led to a decrease in the slope and can be interpreted as evidence of their effect on the diffusion part of the double layer. Its contribution decreases in favor of ion transport. All this, in general, does not contradict the assumption of cell adsorption on the electrode surface. Confocal laser scanning microscopy confirmed this assumption and showed cell adhesion to the surface. The data obtained confirm the importance of bioelectrochemical processes of the studied group of microorganisms in biotechnological processes associated with the leaching of metals from sulfide ores.

Registration of BOD using Paracoccus yeei bacteria isolated from activated sludge.

This work investigated the properties of Paracoccus yeei VKM B-3302 bacteria isolated from activated sludge and immobilized in an N-vinylpyrrolidone-modified poly(vinyl alcohol) matrix. The developed hydrogel formed a network structure to enable the entrapment of microbial cells with their viability and biocatalytic properties preserved, which ensured the technological possibility of replicating expendable biosensor receptor elements. A new ratio of the components for the synthesis selected in this work enabled producing a copolymer of an earlier undescribed chemical structure, which can be efficiently used for immobilization of highly sensitive P. yeei bacteria. A biological oxygen demand (BOD) biosensor with these bacteria and matrix was shown to possess a long-time stability exceeding that described earlier, to have a broad substrate specificity and to exceed approximately tenfold the nearest analogues by its sensitivity and the lower boundary value of 0.05 mg/dm3. The biosensor enabled assays of water samples initially attributed to pure samples (the BOD range, 0.05-5.0 mg/dm3). BOD assays of water samples from various sources showed the use of the receptor element of this composition to enable the data that closely correlated with the standard method (R 2 = 0.9990).

Use of one- and two-mediator systems for developing a BOD biosensor based on the yeast Debaryomyces hansenii.

We investigated the use of one- and two-mediator systems in amperometric BOD biosensors (BOD, biochemical oxygen demand) based on the yeast Debaryomyces hansenii. Screening of nine mediators potentially capable of electron transfer - ferrocene, 1,1'-dimethylferrocene, ferrocenecarboxaldehyde, ferroceneacetonitrile, neutral red, 2,6-dichlorophenolindophenol, thionine, methylene blue and potassium ferricyanide - showed only ferrocene and neutral red to be efficient electron carriers for the eukaryotes studied. Two-mediator systems based on combinations of the investigated compounds were used to increase the efficiency of electron transfer. The developed two-mediator biosensors exceeded their one-mediator analogs by their characteristics. The most preferable two-mediator system for developing a BOD biosensor was a ferrocene-methylene blue combination that ensured a satisfactory long-time stability (43 days), selectivity, sensitivity (the lower limit of the determined BOD5 concentrations, 2.5mg О2/dm3) and speed (assay time for one sample, not greater than 10min) of BOD determination. Analysis of water samples showed that the use of a ferrocene-methylene blue two-mediator system and the yeast D. hansenii enabled registration of data that highly correlated with the results of the standard method (R=0.9913).

[Competition between redox mediator and oxygen in the microbial fuel cell].

The maximal rates and effective constants of 2,6-dichlorphenolindophenol and oxygen reduction by bacterim Gluconobacter oxydans in bacterial fuel cells under different conditions were evaluated. In an open-circuit mode, the rate of 2,6-dichlorphenolindophenol reduction coupled with ethanol oxidation under oxygen and nirogen atmospheres were 1.0 and 1.1 µM s­1 g­1, respectively. In closed-circuit mode, these values were 0.4 and 0.44 µM s­1 g­1, respectively. The initial rate of mediator reduction with the use of membrane fractions of bacteria in oxygen and nitrogen atmospheres in open-circuit mode were 6.3 and 6.9 µM s­1 g­1, whereas these values in closed-circuit mode comprised 2.2 and 2.4 µM s­1 g­1, respectively. The oxygen reduction rates in the presence and absence of 2,6-dichlorphenolindophenol were 0.31 and 0.32 µM s­1 g­1, respectively. The data obtained in this work demonstrated independent electron transfer from bacterial redox centers to the mediator and the absence of competition between the redox mediator and oxygen. The results can make it possible to reduce costs of microbial fuel cells based on activity of acetic acid bacteria G. oxydans.

A yeast co-culture-based biosensor for determination of waste water contamination levels.

Artificial microbial co-cultures were formed to develop the receptor element of a biosensor for assessment of biological oxygen demand (BOD). The co-cultures possessed broad substrate specificities and enabled assays of water and fermentation products within a broad BOD range (2.4-80 mg/dm(3)) with a high correlation to the standard method (R = 0.9988). The use of the co-cultures of the yeasts Pichia angusta, Arxula adeninivorans and Debaryomyces hansenii immobilized in N-vinylpyrrolidone-modified poly(vinyl alcohol) enabled developing a BOD biosensor possessing the characteristics not inferior to those in the known biosensors. The results are indicative of a potential of using these co-cultures as the receptor element base in prototype models of instruments for broad application.

Yeast-based self-organized hybrid bio-silica sol-gels for the design of biosensors.

The methylotrophic Pichia angusta VKM Y-2559 and the oleaginous Cryptococcus curvatus VKM Y-3288 yeast cells were immobilized in a bimodal silica-organic sol-gel matrix comprised of tetraethoxysilane (TEOS), the hydrophobic additive methyltriethoxysilane (MTES) and the porogen polyethylene glycol (PEG). Under carefully optimized experimental conditions, employing basic catalysts, yeast cells have become the nucleation centers for a silica-organic capsule assembled around the cells. The dynamic process involved in the formation of the sol-gel matrix has been investigated using optical and scanning electron microscopic techniques. The results demonstrated the influence of the MTES composition on the nature of the encapsulation of the yeast cells, together with the architecture of the three-dimensional (3D) sol-gel biomatrix that forms during the encapsulation process. A silica capsule was found to form around each yeast cell when using 85 vol% MTES. This capsule was found to protect the microorganisms from the harmful effects that result from exposure to heavy metal ions and UV radiation. The encapsulated P. angusta BKM Y-2559 cells were then employed as a biosensing element for the detection of methanol. The P. angusta-based biosensor is characterized by high reproducibility (Sr, 1%) and operational stability, where the biosensor remains viable for up to 28 days.

Study of polarization properties of fiber-optics probes with use of a binary phase plate.

We conduct a theoretical and experimental study of the distribution of the electric field components in the sharp focal domain when rotating a zone plate with a π-phase jump placed in the focused beam. Comparing the theoretical and experimental results for several kinds of near-field probes, an analysis of the polarization sensitivity of different types of metal-coated aperture probes is conducted. It is demonstrated that with increasing diameter of the non-metal-coated tip part there occurs an essential redistribution of sensitivity in favor of the transverse electric field components and an increase of the probe's energy throughput.

[Bioanode for a microbial fuel cell based on Gluconobacter oxydans inummobilized into a polymer matrix].

Acetic acid bacteria Gluconobacter oxydans subsp. industrius RKM V-1280 were immobilized into a synthetic matrix based on polyvinyl alcohol modified with N-vinylpyrrolidone and used as biocatalysts for the development ofbioanodes for microbial fuel cells. The immobilization method did not significantly affect bacterial substrate specificity. Bioanodes based on immobilized bacteria functioned stably for 7 days. The maximum voltage (fuel cell signal) was reached when 100-130 µM of an electron transport mediator, 2,6-dichlorophenolindophenol, was added into the anode compartment. The fuel cell signals reached a maximum at a glucose concentration higher than 6 mM. The power output of the laboratory model of a fuel cell based on the developed bioanode reached 7 mW/m2 with the use of fermentation industry wastes as fuel.

[Hemodynamic abnormalities in different kind of arteriovenous access for hemodialysis].

On the basis of examination of 35 dialytic patients, who had clinical findings of progressed chronic cardiac insufficiency against the background of the large arteriovenous shunt through existing vascular access, the authors show the complications. The volume of blood circulation along the arteriovenous fistula, which was more than 30% of cardiac output, caused dilatation of heart cavities with cardiac decompensation. If the largest potency to volume remodeling of native proximal arteriovenous fistulas is taken into account, this kind of access could cause hemodynamic abnormalities more often than others (in 22 (62.9%) of patients). The best primary survival was demonstrated by arteriovenous shunts (87.1%) during 2 years. However, long-term survival of native arteriovenous fistulas dramatically outperformed the other kinds of accesses. The choice of constant vascular access for hemodialysis is an important and difficult task in chronic renal disease of V degree patients.

BOD biosensor based on the yeast Debaryomyces hansenii immobilized in poly(vinyl alcohol) modified by N-vinylpyrrolidone.

An amperometric biosensor for assessing the biochemical oxygen demand (BOD) was formed by immobilizing Debaryomyces hansenii VKM Y-2482 yeast cells in poly(vinyl alcohol) modified by N-vinylpyrrolidone. Modification provided for a high sensitivity and stability of the bioreceptor. A high oxidative activity of the receptor element and the absence of any toxic effect of assayed compounds were shown for 34 substrates (alcohols, carbohydrates, carboxylic acids, amino acids, nitrophenols and surfactants) that may occur in wastewaters. Estimates of the measurement range and region of the linear dependence of signals on the BOD level, pH and temperature sensitivities, dependences of signals on concentrations of salts, stability, Michaelis kinetic constants and assay rates were obtained. The BOD values determined by the biosensor in assayed wastewater samples were shown to have a high correlation with those obtained by the standard dilution method.

Strengthening the longitudinal component of the sharply focused electric field by means of higher-order laser beams.

We study the sharp focusing of differently polarized low-order and high-order beams, including Bessel and Laguerre-Gaussian (LG) modes, to compare them using several criteria: the size of a light spot, the intensity ratio of the central peak and sidelobes, and the intensity of the longitudinal electric field component. The experiments performed using the near-field microscopy techniques are in general agreement with the results of the numerical simulation. We have validated the growth of the longitudinal component in the focus for high-order modes at moderate NA=0.6-0.8, and essential lower sidelobes of Bessel modes, in comparison with LG modes.

[Properties of modified amperometric biosensors based on methanol dehydrogenase and Methylobacterium nodulans cells].

The properties of amperometric biosensors based on methanol dehydrogenase (MDH), Methylobacterium nodulans cells, and the ferrocene-modified carbon paste electrode were investigated. It was shown that the addition ofhydroxyapatite (HA) to a carbon paste increased the sensitivity and operating stability of MDH biosensors. The linear range of the electrode was 0.0135-0.5 and 0.032-1.5 mM for methanol and formaldehyde, respectively. The detection limit of methanol and formaldehyde was 4.5 and 11.0 microM, respectively. The loss of activity of the electrode within 10 days of storage in the presence of 2.0 mM KCN did not exceed 12%. Cyanide (10 mM) completely inhibited the sensor responses to formaldehyde (1.0 mM), which allowed for the selective determination of methanol in the presence of formaldehyde. The biosensor based on cells exhibited lower stability and sensitivity toward methanol and formaldehyde; the sensitivity coefficients were 980 and 21 nA/mM, respectively.

[Phenomenon of ischemic steal syndrome in patients with different arteriovenous fistulas for hemodialysis and its surgical correction].

Patients with arteriovenous fistulas (648 cases) were examined for hemodialysis. The ischemic steal syndrome was detected in 47 patients and it was associated with vascular access for hemodialysis. A frequency of given complication was studied with correlation of the type of vascular access. The data of clinical and instrumental examination were described. It was revealed, that the development of steal syndrome was influenced by the blood circulation volume velocity on fistulas. Different reconstructive operations were performed on 42 patients in order to save the access for hemodialysis and eliminate the ischemia. The ligation of fistula was carried out for 5 patients. The survival of arteriovenous fistulas after reconstructive operations was till 46 months during the period of follow-up study.