Investigation of the biological activity of solutions activated electrochemically in a membrane electrolyzer

The biological activity of the catholyte and anolyte of double distilled water was studied in experiments on the germination of wheat grains in the period from March to May. The activity of the solutions, which was characterized by a growth index, was high early in this period, then decreased almost to zero in the middle of the period, and then increased to about the initial value by the end of the period. Throughout, the efficiency of the anolyte of double distilled water generally exceeded the efficiency of the catholyte. Early and late in the period, the stimulatory effect of the anolyte exceeded that of the catholyte by a factor of 5–5.5. The changes in the biological activity of the catholyte and anolyte of double distilled water were also compared with the changes in the biological activity of the catholyte of nutrient medium M9. The stimulatory effect of the catholyte of the nutrient medium was evaluated from the change in the growth of E. coli cells. Early in the period at a cultivation temperature of 20°C, the stimulatory effect determined from the increase in the optical density of the cell suspension in the experiment with respect to a reference value was 55–60%. Next, the stimulatory effect decreased almost to zero in the middle of the period and increased to approximately initial value by the end of the period. It was assumed that the physicochemical mechanisms of action of the catholyte and anolyte of double distilled water on the wheat seed germination and of the catholyte of the nutrient medium on E. coli cell growth are of different nature.     

The causes of the biological action of electrochemically activated solutions by changes in the growth of Escherichia coli cells

To study the causes of the biological effect of electrochemically activated solutions, nutrient growth media M 9 were prepared using catholyte and anolyte solutions containing separate components of the nutrient medium, such as distilled water, phosphate buffer, phosphate buffer with chlorides (NaCl, NH4Cl), and chlorides. The biological activity of different nutrient media was assessed by a comparison with the stimulation or inhibition of the growth of Escherichia coli cells in the catholyte and anolyte of the complete nutrient medium M 9. It was shown that medium M 9 prepared on the catholytes of different initial solutions acquired the stimulating properties only if the initial solution contained salts containing chlorine. The stimulating effect of the initial solution was 18-24%. Electrochemical treatment of solutions containing no chlorides (distilled water, phosphate buffer) and subsequent addition of the components of nutrient medium to exposed solutions had neither a stimulating nor the inhibiting effect on cell growth. The cultivation of cells in a nutrient medium based on the catholyte of preliminarily treated hydrochloric acid showed that it is the presence of chlorine ions in solution during electrolysis that causes the stimulating effect of the nutrient medium based on the catholyte. The formation of oxidizers and the inhibitory effect of the anolyte described previously was also observed if the solution contained chlorine ions during electrolysis. Possible mechanisms of the biological effect of catholytes containing chlorides during electrolysis were discussed.     

Level of hydrogen peroxide in electrochemically activated solutions and study of its effect on growth of Escherichia coli

The formation of hydrogen peroxide in catholytes and anolytes of electrochemically activated solutions: bidistilled water and solutions of sodium chloride and nutrition medium M9 was studied. The concentration of hydrogen peroxide was determined by the method of enhanced chemiluminescence in a system peroxidase-luminol-p-iodophenol. It was shown that the concentration of hydrogen peroxide depends on the ionic content of the solution and varies from a few fractions of a micromole in catholytes of bidistilled water and sodium chloride solutions (10(-5) divided by 10(-2) M) to 20-25 microM in catholytes of medium M9. The concentration of H2O2 in anolytes of various solutions was 15-20 times lower than in the corresponding catholytes and was equal to a few nanomoles in bidistilled water and a few micromoles in medium M9. The biological activity of the catholyte of medium M9 was determined from changes in the growth of E. coli cells. It was found that this catholyte stimulates the cell growth. The stimulating effect was 20-25% and did not change after the decomposition of hydrogen peroxide in the catholyte by catalase. The addition of H2O2 at the corresponding concentration to the inactivated nutrient medium produced no stimulating effect. These data suggest that hydrogen peroxide formed in the catholyte of nutrient medium M9 does not affect its biological activity.     

On the fluorescence of bidistilled water after electrochemical treatment

The short-wave fluorescence of bidistilled water treated in the cathode and anode chambers of two types of electrolysers made from different materials was investigated by fluorescence spectroscopy. It was shown that the electrochemical treatment of water did not induce intrinsic short-wave flyorescence of catholyte and anolyte in the case of quartz glass electrolyser. The increase observed in the intensity of intrinsic short-wavelength fluorescence of bidistilled water in the catholyte and anolyte of the electrolyser made from technical organic glass was caused by microcontaminations released from electrolyser material in the samples.     

Causes of fluorescence in bidistilled water after electrochemical treatment

Fluorescence spectroscopy was used to study the shortwave fluorescence of bidistilled water treated in the cathode and anode chambers of two types of electrolyzers made of different materials. Electrochemical treatment in the quartz glass electrolyzer did not induce intrinsic fluorescence of the anolyte or catholyte. An increase in the shortwave fluorescence of the anolyte and catholyte was observed in the electrolyzer made of Plexiglass, which was probably due to the release of microcontaminants from components of the electrolyzer.     

Stimulation and inhibition of Escherichia coli cell growth during cultivation in the catholyte and anolyte of culture medium

The effect of pretreatment of growth medium M-9 with direct electric current in the cathode and the anode compartments of a diaphragm electrolyzer on the growth of Escherichia coli cells was studied. The cells were cultured separately in the catholyte and the anolyte of the growth medium. The cell growth was registered as a change in optical density of the culture suspension by the method of turbidimetry. It was found that cells grown in the catholyte at a temperature of 37 degrees C yielded a 20-30% increase in amount as compared to the control. No cell growth was observed in the anolyte, and a part of the initial cells were lysed. Possible mechanisms of stimulation and inhibition of cell growth and the reasons of discrepancies in the earlier published data are discussed.     

Nonequilibrium state of electrochemically activated water and its biological activity]

Changes in the physicochemical parameters (pH, redox potential and electroconductivity) of catholyte and anolyte produced by membrane electrolysis of distilled water and dilute (c < 10(-3) M) sodium chloride solutions were studied. The relaxation of these parameters after electrolysis and the influence of catholyte and anolyte on the growth of roots of Tradescantia viridis grafts, the development of duckweed, and the motive activity of infusoria Spirostomum ambiguum were investigated. It was found that the anolyte of distilled water stimulated development of these biological objects. The direction of shift of physicochemical parameters of catholyte and anolyte from equilibrium values and the type of their biological activity (stimulation or inhibition) depend on salt concentration in initial solution. Barbotage of initial distilled water with argon or nitrogen leads to a greater decrease in the redox potential of catholyte during electrolysis. The physicochemical parameters relax to equilibrium values, and the biological activity of catholite and anolyte decreases with time and practically disappears by the end of the day. It was found that the oxidation of reducing agent by atmospheric oxygen is not the sole cause of the relaxation of catalyte redox potential. The increase in the ionic strength of catholite and anolyte by the addition of concentrated sodium chloride after electrolysis decreases the rate of redox potential relaxation several times. The redox potential can be maintained for long periods by freezing.     

Effect of Subsoiling in Fallow Period on Soil Water Storage and Grain Protein Accumulation of Dryland Wheat and Its Regulatory Effect by Nitrogen Application

To provide a new way to increase water storage and retention of dryland wheat, a field study was conducted at Wenxi experimental site of Shanxi Agricultural University. The effect of subsoiling in fallow period on soil water storage, accumulation of proline, and formation of grain protein after anthesis were determined. Our results showed that subsoiling in fallow period could increase water storage in the 0–300 cm soil at pre-sowing stage and at anthesis stage with low or medium N application, especially for the 60–160 cm soil. However, the proline content, glutamine synthetase (GS) activity, glutamate dehydrogenase (GDH) activity in flag leaves and grains were all decreased by subsoiling in fallow period. In addition, the content of albumin, gliadin, and total protein in grains were also decreased while globulin content, Glu/Gli, protein yield, and glutelin content were increased. With N application increasing, water storage of soil layers from 20 to 200 cm was decreased at anthesis stage. High N application resulted in the increment of proline content and GS activity in grains. Besides, correlation analysis showed that soil storage in 40–160 cm soil was negatively correlated with proline content in grains; proline content in grains was positively correlated with GS and GDH activity in flag leaves. Contents of albumin, globulin and total protein in grains were positively correlated with proline content in grains and GDH activity in flag leaves. In conclusion, subsoiling in fallow period, together with N application at 150 kg·hm⁻², was beneficial to increase the protein yield and Glu/Gli in grains which improve the quality of wheat.     

Molecular cloning and characterization of an inorganic pyrophosphatase from barley

A cDNA clone with sequence homology to soluble inorganic pyrophosphatase (IPPase) was isolated from a library of developing barley grains. The protein encoded by this clone was produced in transgenic Escherichia coli, and showed IPPase activity. In nondormant barley grains, the gene appeared to be expressed in metabolically active tissue such as root, shoot, embryo and aleurone. During imbibition, a continuous increase of the steady state mRNA level of IPPase was observed in embryos of non-dormant grains. In the embryos of dormant grains its production declined, after an initial increase. With isolated dormant and nondormant embryos, addition of recombinant IPPase, produced by E. coli, enhanced the germination rate. On the other hand, addition of pyrophosphate (PPi), substrate for this enzyme, appeared to reduce the germination rate. A role for this IPPase in germination is discussed.     

Induction of phenolic compounds in wheat (Triticum aestivum L.) tissue cultures by streptomycin

The tissue cultures of wheat (Triticum aestivum L.) were induced from the mature embryos (explants) of the dry grains and grown on MS medium containing kinetin (0.1 mg/1) and 2,4 D (1.0 mg/l). The cultures were incubated for two weeks at (25+/-2) degrees C under a light/dark regime (16 h light daily). The formed calli were subcultured at the beginning of the stationary growth phase (15 days) with fresh MS medium containing 0, 5, 10, 25, 50, 100, 150 mg/l streptomycin elicitor and maintained for two weeks for three subcultures. A significant increase in phenylalanine ammonia lyase (PAL) activity coincided with the increase of the total phenolic compounds after elicitation with streptomycin. Maximum induction was recorded during the first two weeks, then gradually declined during the rest of the experimental period, but the values attained were still markedly higher than that of the control. The endogenous cinnamic acid content was also increased significantly with the increase in PAL activity making about 2-18% of the total phenolic acids. The growth and accumulation of phenolic compounds were inversely related. However, accumulation of phenolic compounds became limited for growth of wheat tissue culture especially during the long term cultivation.     

Stabilization of pH gradients in buffer electrofocusing on polyacrylamide gel.

pH gradients in buffer electrofocusing on polyacrylamide gel (BEF) have been stabilized for at least 100 hr, 25°C, by replacing the strongly acidic and basic anolyte and catholyte with isoelectric buffers identical to the terminal constituents of the pH gradient and gel. Such stabilization leads to a constant pI position of an electrofocused protein. No stabilization of pH gradients is achieved under otherwise identical conditions when strongly acidic and basic anolyte and catholyte are used.     

Immunomodulating properties of bidistilled modified water

The effect of bidistilled modified water obtained by treating bidistilled water with static electric field in the cathode compartment of a diaphragm electrolyzer on the production of tumor necrosis factor and activity of natural killer cells of mice was studied. It was found that bidistilled modified water added to cultured murine macrophages increases the tumor necrosis factor production 2.5-3-fold. A similar degree of stimulation of tumor necrosis factor production was observed after peroral administration of bidistilled modified water for 10-25 days. The activity of natural killer cells under these conditions increased by 150%. The stimulation of these immunity system components responsible for the resistance of the organism to tumors led to a suppression of tumor growth in tumor-bearing mice receiving bidistilled modified water.     

Surfactant production by the Rhodococcus erythropolis sH-5 bacterium grown on various carbon sources

It has been shown that the Rhodococcus erythropolis sH-5 strain can produce surfactants associated and not associated with the cell wall. Their content depends on medium composition, the nature of the carbon source, and oxygen supply. The highest biosurfactant (bioSF) yield is achieved by growing R. erythropolis sH-5 in medium with 2% kerosene at neutral pH. It has been found that the bioSF yield and emulsification index for various hydrocarbons depend on the kind of the nitrogen source used by the bacterium, increasing with replacement of KNO3 by NaNO3. The yields of biomass and bioSF in R. erythropolis depend on growth temperatures (max at 30 degrees C) but not on water quality (bidistillate, catholyte, or anolyte). It has been found that sH-5 produces more cell-associated bioSF than extracellular species.     

Hydrolysis of the bovine insulin B-chain in bidistilled modified water

It was shown that bidistilled modified water substantially enhances the hydrolysis of the peptide the bovine insulin B-chain. The exposure of the peptide to bidistilled modified water for 20 hours at room temperature leads to an almost complete hydrolysis of its molecule into fragments that differ from the initial molecule in elution time from a column in high-performance liquid chromatography.     

绿色木霉在啤酒糟基质上产纤维素酶条件的优化

对绿色木霉接种到啤酒糟固态发酵产纤维素酶的培养基和培养条件进行优化,考察发酵物料起始含水量、发酵时间、起始pH值等发酵条件,以及啤酒糟培养基中添加麸皮、氮源种类对产酶的影响。结果表明,以啤酒糟为发酵基质接种绿色木霉生产纤维素酶是可行的。经单因素和正交试验获得最适固态发酵的培养条件为:起始pH 5~6,培养温度28~30℃,发酵4 d;最佳发酵培养基组合为:麸皮比例30%,培养基起始含水量50%,(NH₄)₂SO₄添加量为2.0%~2.5%。     

一氧化氮对渗透胁迫下小麦种子萌发及其活性氧代谢的影响

一氧化氮供体硝普钠(Sodium nitroprusside,SNP)能明显地促进渗透胁迫下小麦(Triticum aestivum L.)种子萌发、胚根和胚芽伸长,提高萌发过程中淀粉酶和内肽酶的活力,加速贮藏物质的降解;胁迫解除后,仍能使种子维持较高的活力.此外,SNP还能显著诱导渗透胁迫下CAT、APX活力的上升和脯氨酸含量积累,抑制LOX活力,从而提高渗透胁迫下小麦种子萌发过程中抗氧化能力.进一步研究还发现,SNP诱导切胚半粒小麦种子萌发早期(6h)的淀粉酶活力上升可能与GA3无直接关系.     

一氧化氮对渗透胁迫下小麦种子萌发及其活性氧代谢的影响

一氧化氮供体硝普钠(Sodium nitroprusside,SNP)能明显地促进渗透胁迫下小麦(Triticum aestivum L.)种子萌发、胚根和胚芽伸长,提高萌发过程中淀粉酶和内肽酶的活力,加速贮藏物质的降解：胁迫解除后,仍能使种子维持较高的活力。此外,SNP还能显著诱导渗透胁迫下CAT、APX活力的上升和脯氨酸含量积累,抑制LOX活力,从而提高渗透胁迫下小麦种子萌发过程中抗氧化能力。进一步研究还发现,SNP诱导切胚半粒小麦种子萌发早期(6h)的淀粉酶活力上升可能与GA3无直接关系。     

Respiratory activity during germination of seeds of Cicer arietinum L. II. Effect of some metabolic inhibitors and mitochondrial activities

Changes in respiration rate during the early period of germination of Cicer arietinum L. seeds were studied. Three phases were distinguished. During phases I and II the RQ was higher than unity. Sodium fluoride showed no effect on gaseous exchange during phase I while in phases II and III a progressive increase in the inhibition of the respiratory activity was obtained. Iodoacetate inhibited oxygen uptake and carbon dioxide evolution from the beginning of germination. The activities of mitochondrial enzymes increased during the germination period, especially after the first 24 hours. The respiratory control and ADP/O ratio of the isolated mitochondria increased gradually during germination, reaching maximum values at 48 hours, lower, however than the expected theoretical ones.     

Development of a population-based threshold model of conidial germination for analysing the effects of physiological manipulation on the stress tolerance and infectivity of insect pathogenic fungi

Entomopathogenic fungi are being used as biocontrol agents of insect pests, but their efficacy can be poor in environments where water availability is reduced. In this study, the potential to improve biocontrol by physiologically manipulating fungal inoculum was investigated. Cultures of Beauveria bassiana, Lecanicillium muscarium, Lecanicillium longisporum, Metarhizium anisopliae and Paecilomyces fumosoroseus were manipulated by growing them under conditions of water stress, which produced conidia with increased concentrations of erythritol. The time-course of germination of conidia at different water activities (water activity, aw) was described using a generalized linear model, and in most cases reducing the water activity of the germination medium delayed the onset of germination without affecting the distribution of germination times. The germination of M. anisopliae, L. muscarium, L. longisporum and P. fumosoroseus was accelerated over a range of aw levels as a result of physiological manipulation. However, the relationship between the effect of physiological manipulation on germination and the osmolyte content of conidia varied according to fungal species. There was a linear relationship between germination rate, expressed as the reciprocal of germination time, and aw of the germination medium, but there was no significant effect of fungal species or physiological manipulation on the aw threshold for germination. In bioassays with M. anisopliae, physiologically manipulated conidia germinated more rapidly on the surface of an insect host, the melon cotton aphid Aphis gossypii, and fungal virulence was increased even when relative humidity was reduced after an initial high period. It is concluded that physiological manipulation may lead to improvements in biocontrol in the field, but choice of fungal species/isolate will be critical. In addition, the population-based threshold model used in this study, which considered germination in terms of physiological time, also called hydrotime, could have general application in mycology and environmental microbiology.