The foliar uptake of dimethomorph into cucumber was assessed by spray application of aqueous dimethomorph solution containing fatty alcohol ethoxylate (FAE) or fatty acid alkyl ester as activator adjuvants. Afterward, the possible mechanism of action of FAE on foliar penetration of active ingredient was suggested by speculating on the effect of lipophile and hydrophile of FAEs. The amount of absorbed dimethomorph induced by polyoxyethylene mono-9-octadecenyl ether (6 moles ethylene oxide, $C_{18=9}E_6$) was linearly related to the concentrations of surfactant as well as dimethomorph in spray solution, suggesting that it is simply a diffusion phenomenon of the solute molecule through a cuticular membrane from leaf surface. Octadecanol attached to FAE was most effective lipophile on the leaf penetration of dimethomorph. And, the more ethylene oxide had the polyoxyethylene chain of FAE up to 20 moles, the higher the uptake rate was. Therefore, the role of lipophile of FAE on dimethomorph penetration to cucumber leaf, probably, is to modify the physico-chemical properties of cuticular membrane to be permeable to dimethomorph, and the polyoxyethylene chain having less than 20 moles ethylene oxide, which is moderately permeable to cuticular membrane by its molar volume, is to let the physically-modified cuticular membrane to be maintained for a longtime.
Journal of Physiology & Pathology in Korean Medicine
/
v.27
no.1
/
pp.112-117
/
2013
We studied the modulation of pacemaker activities by Samchulkunbi-tang (SCKB) in cultured interstitial cells of Cajal (ICC) from murine small intestine with the whole-cell patch-clamp technique. Externally applied SCKB produced membrane depolarization in the current-clamp mode. The pretreatment with $Ca^{2+}$-free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum, abolished the generation of pacemaker potentials and suppressed the SCKB-induced action. The application of flufenamic acid (a nonselective cation channel blocker) abolished the generation of pacemaker potentials by SCKB. However, the application of niflumic acid (a chloride channel blocker) did not inhibit the generation of pacemaker potentials by SCKB. In addition, the membrane depolarizations were inhibited by not only GDP-${\beta}$-S, which permanently binds G-binding proteins, but also U-73122, an active phospholipase C inhibitor. These results suggest that SCKB modulates the pacemaker activities by nonselective cation channels and external $Ca^{2+}$ influx and internal $Ca^{2+}$ release via G-protein and phospholipase C-dependent mechanism. Therefore, the ICC are targets for SCKB and their interaction can affect intestinal motility.
Rhamnose-rich (RROP-s) and fucose-rich (FROP-s) oligo-and polysaccharides were prepared and extensively characterised by physical and chemical procedures [1,2] and compared to L-fucose. Their biological properties were then studied on human skin fibroblast cell cultures, human skin explant cultures and on hairless rat skin, using a variety of cell-biological, biochemical and computerised morphometrical procedures. Among the most important properties we could establish, the following are of particular interest for the tretment and prevention of age-dependent modifications of human skin (loss of skin-tissue, cells and matrix, wrinkle formation and others) : stimulation of cell proliferation (by $^3$[H]-thymidine incorporation and the MTT test), scavenging of reactive oxygen species (ROS) using several different procedures, and protease (MMP-2 and MMP-9) down-regulation. A topical preparation, using RROP-s and FROP-s, and/or L-fucose, was shown to increase cell proliferation, dermal matrix synthesis, efficient scavenging of ROS-s and to increase also the thickness of dermal tissue when applied for 4 weeks on hairless rat skin, accompanied by the densification of collagen bundles as well as by an increase of elastin synthesis. Using fluorescent labeled FROPs, it could be shown that these oligosaccharides react with cell-membrane receptors and especially with the elastin-laminin-receptor and the fucose-mannose receptor, but they penetrate also in the cell nucleus, suggesting the possibility of a direct action on the regulation of gene expression. When applied to the human skin of a team of voluntary women encompassing all age-groups, the efficiency of FROP-containing preparation could be confirmed using indentometry and computerised evaluation of skin micro-relief, as well as evaluation of periorbital wrinkles. It appears therefore that these preparations correspond to all the requirements of active anti-aging principles.
Various fermented foods were screened in search of food-grade bacteria that produce bacteriocins active against Gram-negative pathogens. An isolate from a mold-ripened cheese presented antibacterial activity against Gram-positive and Gram-negative bacteria. The most active isolate was identified as Lactobacillus casei by a biochemical method, ribotyping, and membrane lipid analysis, and was designated as OSY-LB6A. The cell extracts of the isolate showed inhibition against Escherichia coli p220, E. coli O157, Salmonella enerica serovar Enteritidis, Salmonella Typhimurium, and Listeria monocytogenes. The antibacterial nature of the cell extract from the isolate was confirmed by eliminating the inhibitory effects of acid, hydrogen peroxide, and lytic bacteriophages. The culture supernatant and cell extract retained antibacterial activity after heating at $60{\sim}100^{\circ}C$ for $10{\sim}20$ min. The activity of the cell extract from Lb. casei was eliminated by pronase and lipase. Finally, the cell extract showed a bactericidal mode of action against E. coli in phosphate buffer solution, but it was bacteriostatic in broth medium and food extracts.
Plant mitochondria, irradiated with blue-colored $sunlight(350{\sim}500nm)$ under aerobic and anaerobic conditions, were assayed as to the electron transfer activity of respiratory enzyme system, and compared with those irradiated with orange-colored light(white sunlight minus blue-colored light). The respiratory activity of mitochondria was most seriousely inhibited by illumination with blue-colored light under aerobic condition. Deaeration of mitochondrial suspension resulted in substantial decrease of the photoinhibition by blue-colored light. Meanwhile, orange-colored light demonstrated much less effectiveness-almost ineffectiveness-in causing the inhibition of mitochondrial respiration system. The results of enzymatic assay revealed a strong possibility that FMN in NDH and heme group at least in cytochrome c oxidase, but not FAD in SDH, are the photodynamic sensitizers in mitochondrial inner membrane. Also worthwhile to note is the significant difference from the others of SDH in its photoinhibitory response to the light quality of visible light; that the inhibition of SDH by irradiation was not affected by atmospheric condition and that orange-colored light gave rise to considerable extents of inhibition to the enzyme. This observation was tentatively interpreted in terms of photosensitized reaction not involving molecular oxygen possibly catalyzed by Fe-S centers in the enzyme. The superoxide production and the membrane peroxidation of mitochondria under various treatments also indicated that there was blue-light photodynamic reaction in mitochondria involving active oxygens.
The aim of this study was to determine if hydrogen peroxide ($H_2O_2$) generated by glucose oxidase (GO) induces apoptosis or necrosis of BJAB cells and which radical is the direct mediator of cell death. We found that GO produced $H_2O_2$ continuously in low concentrations, similar to in vivo conditions, and decreased proliferation and cell viability in a dose-dependent manner. The GO-mediated cytotoxicity resulted from apoptosis, and was confirmed by monitoring the cells after H33342/Annexin V/propidium iodide staining. Decreases of mitochondrial membrane potential and intracellular glutathione level were found to be critical events in the $H_2O_2$-mediated apoptosis. Additional experiments revealed that $H_2O_2$ exerted its apoptotic action through the formation of hydroxyl radicals via the Fenton rather than the Haber-Weiss reaction. Moreover, intracellular redox-active iron, but not copper, participated in the $H_2O_2$-mediated apoptosis.
Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.
The fatty acid compositions of phospholipids extracted from leaves and leaf mitochondria, which were sampled from several horicultural plants grown under blue color-deficient sunlight (BCDS), were determined and compared with those from plants grown under natural white colored sunlight(WCS). It was found that the mitochondria isolated from plants grown under BCDS contained phospholipid whose degree of unsaturation in unit of number of double bonds per lipid molecule was remarkably higher than that from plants grown under WCS, the relative increment being $8{\sim}49%$. This was significantly larger than the relative increment, $4{\sim}8%$ for total phospholipid extracted from whole leaves grown under BCDS campared to WCS. This observation demonstrated that the blue light effect of sunlight on the chemical property of cellular membranes, as long as it was concerned with fatty acid composition, arose mainly at the mitochondrial membrane. Also observing that the degree of unsaturation of mitochondrial phospholipid was much lower than that of total phospholipid, it was interpreted that this was the consequence of rather active oxidative destruction of lipid-fatty acid components occuring in mitochondrial membrane by the reactive oxygen species, especially superoxide($O_2-$), which was known to be produced in mitochondrial inner membrane through the side reactions of the respiratory electron transport chain and also probably through the photosensitized reaction involving oxygen induced by blue colored light. Thus, it may be tentatively concluded that the extent of photosensitization in mitochondrial membrane could be considerably reduced under BCDS resulting in lowering of the $O_2-$ level in the respirating organelle The possible involvement of photodynamic action in membrane oxidation was also indicated by the fact that the typical fat-soluble antioxidant, ${\alpha}-tocopherol$, was found to be contained on a higher level in leaves under BCDS than those under WCS.
The effects of various inotropic interventions on the shape of the steady state length tension relation and the length-dependent activation process in cardiac muscle were studied. The influence of inotropic interventions upon the action potential was also observed. The range of varying muscle length was from the optimal length$(l_{max})$, where the active tension production is maximal, to 0.85 $l_{max}$. Changes in stimulus frequency or in external bathing Ca concentration constituted the inotropic interventions in this experiment. The papillary muscles were isolated from the rabbit right ventricles and perfused with $HCO-_3\;-buffered$ normal Tyrode solution which was aerated with $3%\;CO_2-97%\;O_2$ mixed gas and kept at $35^{\circ}C$. Resting Passive tension at $l_{max}$ was approximately 30% of the total tension and appeared from the muscle length of 0.90 $l_{max}$. The effect of stimulus frequency on the steady state level of developed tension was: As the stimulus frequency was increased from 0.1 to 0.5 Hz, there was little change in developed tension. As the frequency was increased further, to a value of about 3 Hz, tension increased steeply. Further increase of the frequency to 5 Hz had little additional effect on the developed tension. The length-tension curves for isometric peak tension became more steeper with the degree of potentiation by inotropic interventions. The relative steepness of the normalized length-tension curves where tension production was expressed as a percentage of maximal tension developed at $l_{max}$, varied inversely with the level of inotropic state and these curves were not superimposable one another. Thus at the stimulus frequency of 2 Hz or at the external Ca concentration of 8 mM, the relative decline in the developed tension for a given change in muscle length was considerably less than the decline observed at the frequency of 0.5 Hz or at the concentration of 2 mM Ca. Action potential duration was prolonged significantly as the frequency increased from 0.2 to 2 Hz, and this change in action potential duration was not observable on the changes in muscle length. There was a tendency of the hyperpolarization of membrane potential when the muscle length was shortened from $l_{max}$ to 0.95 $l_{max}$. These results support the hypothesis that there is a length-dependence of the activation process.
Rakitin, Andrey L.;Ermakova, Alexandra Y.;Ravin, Nikolai V.
Journal of Microbiology and Biotechnology
/
v.25
no.9
/
pp.1476-1484
/
2015
Three endoxylanase-encoding genes from the moderately themophilic chemoorganotrophic bacterium Melioribacter roseus were cloned and expressed in Escherichia coli. Genes xyl2091 (Mros_2091) and xyl2495 (Mros_2495) encode GH10 family hydrolases, whereas xyl2090 (Mros_2090) represents the GH30 family. In addition to catalytic domains, Xyl2090 and Xyl2091 contain carbohydrate-binding modules that could facilitate their binding to xylans and Por sorting domains associated with the sorting of proteins from the periplasm to the outer membrane, where they are covalently attached. Recombinant endoxylanase Xyl2495 exhibited a high specific activity of 1,920 U/mg on birchwood xylan at 40℃. It is active at low temperatures, exhibiting more than 30% of the maximal activity even at 0℃. Endoxylanases Xyl2090 and Xyl2091 have lower specific activities but higher temperature optima at 80℃ and 65℃, respectively. Analysis of xylan hydrolysis products revealed that Xyl2090 generates xylo-oligosaccharides longer than xylopentaose. Xylose and xylobiose are the major products of xylan hydrolysis by the recombinant Xyl2091 and Xyl2495. No activity against cellulose was observed for all enzymes. The presence of three xylanases ensures efficient xylan hydrolysis by M. roseus. The highly processive "free" endoxylanase Xyl2495 could hydrolyze xylan under moderate temperatures. Xylan hydrolysis at elevated temperatures could be accomplished by concerted action of two cell-bound xylanases; Xyl2090 that probably degrades xylans to long xylo-oligosaccharides, and Xyl2091 hydrolyzing them to xylose and xylobiose. The new endoxylanases could be useful for saccharification of lignocellulosic biomass in biofuels production, bleaching of paper pulp, and obtaining low molecular weight xylooligosaccharides.
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