Effect of diethylsulphoxide on growth,survival and ion exchange of Escherichia coli

AIMS: To study the effect of diethylsulphoxide (DESO) on Escherichia coli growth, survival and ionic exchange in comparison with dimethylsulphoxide (DMSO). METHODS AND RESULTS: Bacterial survival was estimated by counting colony-forming units and by the most probable number (five-tube) technique; the K+ and H+ transport and H(2) formation were determined electrochemically. Diethylsulphoxide at concentrations between 0.01 and 0.5% (w/v) stimulated and above 5% decreased the anaerobic growth rate and survival. 2H+ : K+ exchange and H(2) formation were lost at 5% DESO. At 0.05% DESO the kinetic characteristics of H+ : K+ exchange and H(2) formation were typical for Delta micro (H(+)) -dependent TrkA uncoupled with F(0)F(1) under respiration. CONCLUSIONS: Diethylsulphoxide at low concentrations serves as an electron acceptor for an anaerobic respiratory chain stimulating bacterial growth and survival through the modulation of H+ : K+ exchange and H(2) formation activity. The effects of DESO were more pronounced than those of DMSO. SIGNIFICANCE AND IMPACT OF THE STUDY: Diethylsulphoxide determines essential biological and therapeutic properties that make its application preferable.     

Stimulation of the production of hypericins by mannan in Hypericum perforatum shoot cultures

Shoot organ cultures were established from callus derived from anthers of Hypericum perforatum flowers and the effect of elicitors on hypericin and pseudohypericin production in shoot organ cultures was investigated. Mannan stimulated pseudohypericin production up to four fold (0.82 mg/g dry wt) and hypericin production up to two fold (0.04 mg/g dry wt.) beta-1,3-glucan and pectin slightly stimulated pseudohypericin production (ca. two fold), but had no effect on hypericin production. On the other hand, yeast extract showed no stimulatory effect, on either hypericin or pseudohypericin production.     

Electrophysiological and Pharmacological Analysis of Neuronal Activity in the Midbrain Medial Raphe Nucleus after Acute Immobilization Stress

In albino rats anesthetized with urethane, most medial raphe neurons are characterized by a low rate of the discharges. Acute immobilization stress resulted in a significant enhancement in the number of regularly discharging neurons. After immobilization, the number of neurons with a low-rate impulsation decreased, while the number of neurons with intensive firing increased. Treatment with fluoxetine enhanced the number of regularly firing neurons more than three times, and a predominant part of the neurons demonstrated a high level of background activity. It should be supposed that a significant part of the neurons with high-rate discharges are serotonergic, because their number also increased after fluoxetine administration.     

The Effect of Cork Pieces on Pseudohypericin Production in Cells ofHypericum perforatumShoots

The stimulating effect of cork pieces on hypericin and pseudohypericin biosyntheses was studied in cells of shoots regenerated from the callus cultures of St. John's wort (Hypericum perforatumL.). The addition of the cork matrix slightly stimulated shoot growth and enhanced pseudohypericin biosynthesis about threefold (to 0.4 mg/g dry wt). Pseudohypericin production increased proportionally with the amount of cork material added (from 1 to 4 mg/ml of growth medium). Further increase in the amount of cork pieces inhibited both pseudohypericin production and shoot growth. Organic and aqueous extracts of cork pieces did not affect the production of these substances.     

Copper (II) Ions Affect <Emphasis Type="Italic">Escherichia coli</Emphasis> Membrane Vesicles’ SH-Groups and a Disulfide-Dithiol Interchange Between Membrane Proteins

The SH-groups in Escherichia coli membrane vesicles, prepared from cells grown in fermentation conditions on glucose at slightly alkaline pH, have a role in the F0F1-ATPase operation. The changes in the number of these groups by ATP are observed under certain conditions. In this study, copper ions (Cu2+) in concentration of 0.1 mM were shown to increase the number of SH-groups in 1.5- to 1.6-fold independent from K+ ions, and the suppression of the increased level of SH-groups by ATP was determined for Cu2+ in the presence of K+. Moreover, the increase in the number of SH-groups by Cu2+ was absent as well as the inhibition in ATP-dependent increasing SH-groups number by Cu2+ lacked when vesicles were treated with N-ethylmaleimide (NEM), specific thiol-reagent. Such an effect was not observed with zinc (Zn2+), cobalt (Co2+), or Cu+ ions. The increased level of SH-groups was observed in the hycE or hyfR mutants with defects in hydrogenases 3 or 4, whereas the ATP-dependent increase in the number of these groups was determined in hycE not in hyfR mutants. Both changes in SH-groups number disappeared in the atp or hyc mutants deleted for the F0F1-ATPase or hydrogenase 3 (no activity of hydrogenase 4 was detected in the hyc mutant used). A direct effect of Cu2+ but not Cu+ on the F0F1-ATPase is suggested to lead to conformational changes or damaging consequences, increasing accessible SH-groups number and disturbing disulfide-dithiol interchange within a protein-protein complex, where this ATPase works with K+ uptake system or hydrogenase 4 (Hyd-4); breaks in disulfides are not ruled out.     

Effect of Various Elicitors on Lignan Biosynthesis in Callus Cultures of Linum austriacum

Effects of elicitors (mannan, -1,3-glucan, and ancymidol) on the activity of several key enzymes participating in lignan biosynthesis were studied in Linum austriacum L. cell cultures. The activities of L-phenylalanine ammonia-lyase, polyphenoloxydase, tyrosine ammonia-lyase, soluble phenoloxidase, and membrane-bound and soluble oxidases were assayed. The elicitors under study affected various steps in the metabolic pathway of lignan biosynthesis. Elevated enzyme activity accompanied an elicitor-enhanced synthesis of podophyllotoxins and peltatins.     

Redox sensing by Escherichia coli: effects of dithiothreitol, a redox reagent reducing disulphides, on bacterial growth

Escherichia coli is able to grow with a high rate under anaerobic conditions upon decrease in redox potential (E(h)) both either in slightly alkaline (pH 7.5) or acidic (pH 5.5) medium. Upon transition of E. coli MC4100 culture to stationary growth phase a decrease in E(h) from the positive values of +120 to +160 mV to the negative ones of -380 to -550 mV, and the H(2) production are observed at various pH. A redox reagent dl-dithiothreitol (DTT) in a concentration of 3mM reduces E(h) to the negative values, and increases a latent (lag) growth phase duration, as well as delays a logarithmic growth phase independently of pH. At alkaline and acidic pH the changes in membrane potential (DeltaPsi) are observed in the presence of 3mM DTT. K(+) uptake is recovered. At pH 5.5 the H(2) production is suppressed by DTT only in a higher concentration of 10 mM. The results suggest DTT effects that are in addition to the effects of E(h). The mechanism of DTT action on bacterial growth might be intermediated through thiol group modulation of the membrane proteins, which is reflected as the generation of DeltaPsi as well as K(+) accumulation and the activity of the membrane-associated enzymes.     

Regulation of isoflavone production in hydroponically grown Pueraria montana (kudzu) by cork pieces, XAD-4, and methyl jasmonate

A mini-hydroponic growing system was employed for seedlings of kudzu vine (Pueraria montana) and contents of isoflavones (daidzein, genistein, daidzin, genistin, and puerarin) from shoot and root parts of seedlings were analyzed quantitatively. In addition, exogenous cork pieces, polymeric adsorbent, XAD-4, and universal elicitor, methyl jasmonate (MeJA), were used to regulate the production of these isoflavones. It was shown that cork pieces up-regulate the production of daidzein and genistein up to seven- and eight-fold greater than the levels obtained for control roots. In contrast, levels of glucosyl conjugates, daidzin and genistin, decrease up to five- and eight-fold, respectively. Cork treatment also induces the excretion of the root isoflavone constituents into the growth medium. Minimal levels of isoflavones are absorbed by the cork pieces. XAD-4 stimulates the production of glucosyl conjugates, daidzin and genistin, in root parts about 1.5-fold greater than that obtained in control roots. These are the highest amounts of daidzin and genistin that are observed (5.101 and 6.759 mg g⁻¹ dry weight, respectively). In contrast to these two adsorbents, MeJA increases the accumulation of isoflavones in shoot rather than in root parts of seedlings, about three- to four-fold over control levels, with the exception of genistein. These studies reveal new observations on the regulation of isoflavone production in hydroponically grown Pueraria montana plants by two adsorbents (cork pieces and XAD-4) and MeJA elicitor.     

Brassinolide interacts with auxin and ethylene in the root gravitropic response of maize (Zea mays)

This study examines how brassinolide (BL) and ethylene interact in the gravitropic response mechanism of maize ( Zea mays ) primary roots. When applied exogenously, ethylene increases the rate of gravitropic curvature in a dose-dependent manner. This effect of ethylene was confirmed by the fact that AVG, a specific action inhibitor of ACC synthase, reduces the gravitropic curvature in the presence and absence of BL. Since AVG did not inhibit BL-increased gravitropic curvature completely, we investigated the possibility that BL may act on the gravitropic response by ways other than simply through enhanced ethylene production. We show that BL exhibits some of its stimulatory effect in the absence of ethylene. In addition, BL reduces the presentation time and lag period for the gravitropic response, whereas ethylene increases them. One possible mechanism of such action is that BL affects protein kinase activity, since the protein kinase inhibitors, staurosporine and H89, reduce BL-increased gravitropic curvature. In summary, BL is involved in the gravitropic response in maize primary roots via ethylene production, but it acts in a way that differs somewhat from that of ethylene.     

Elongation and gravitropic responses of Arabidopsis roots are regulated by brassinolide and IAA

Exogenously applied brassinolide (BL) increased both gravitropic curvature and length of primary roots of Arabidopsis at low concentration (10(-10) M), whereas at higher concentration, BL further increased gravitropic curvature while it inhibited primary root growth. BRI1-GFP plants possessing a high steady-state expression level of a brassinosteroid (BR) receptor kinase rendered the plant's responses to gravity and root growth more sensitive, while BR-insensitive mutants, bri1-301 and bak1, delayed root growth and reduced their response to the gravitropic stimulus. The stimulatory effect of BL on the root gravitropic curvature was also enhanced in auxin transport mutants, aux1-7 and pin2, relative to wild-type plants, and increasing concentration of auxin attenuated BL-induced root sensitivity to gravity. Interestingly, IAA treatment to the roots of bri1-301 and bak1 plants or of plants pretreated with a BL biosynthetic inhibitor, brassinazole, increased their sensitivity to gravity, while these treatments for the BL-hypersensitive transgenic plants, BRI1-GFP and 35S-BAK1, were less effective. Expression of a CYP79B2 gene, encoding an IAA biosynthetic enzyme, was suppressed in BL-hypersensitive plant types and enhanced in BL-insensitive or -deficient plants. In conclusion, our results indicate that BL interacts negatively with IAA in the regulation of plant gravitropic response and root growth, and its regulation is achieved partly by modulating biosynthetic pathways of the counterpart hormone.