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Biomedical subjects

Y Henis

Publications and source records attributed to Y Henis.

At least 19 recordsLinked to original sources

Evidence for cytochrome P-450 and P-450-mediated benzo(a)pyrene hydroxylation in the white rot fungus Phanerochaete chrysosporium.

The presence of cytochrome P-450 and P-450-mediated benzo(a)pyrene hydroxylase activity in both microsomal and soluble fractions of the white rot fungus Phanerochaete chrysosporium was shown. The reduced carbon monoxide difference spectrum showed maxima at 448-450 and 452-454 nm for microsomal and cytosolic fractions, respectively. Both P-450 fractions produced a Type I substrate binding spectrum on addition of benzo(a)pyrene. Activity for benzo(a)pyrene hydroxylation was NADPH dependent and inhibited by carbon monoxide. Km values for activity showed a difference between the cellular fractions with a Km of 89 microM for microsomal P-450 and 400 microM for cytosolic P-450. The Vmax values observed were 0.83 nmol min-1 (nmol microsomal P-450)-1 and 0.4 nmol min-1 (nmol cytosolic P-450)-1. The results indicate that P-450-mediated benzo(a)pyrene hydroxylase activity could play a role in xenobiotic transformation by this fungus beside the known ligninolytic exocellular enzymes.

Basidiomycota↗

Manganese-enhanced biotransformation of atrazine by the white rot fungus Pleurotus pulmonarius and its correlation with oxidation activity.

Manganese enhanced atrazine transformation by the fungus Pleurotus pulmonarius when added to a liquid culture medium at concentrations of up to 300 microM. Both N-dealkylated and propylhydroxylated metabolites accumulated in the culture medium, with the former accumulating to a greater extent than did the latter. Lipid peroxidation, oxygenase and peroxidase activities, and the cytochrome P-450 concentration increased. In addition, an increase in the spectral interactions between atrazine and components in the cell extract was observed. Antioxidants, mainly nordihydroguaiaretic acid, which inhibits lipoxygenase, peroxidase, and P-450 activities, and piperonyl butoxide, which inhibits P-450 activity, inhibited atrazine transformation by the mycelium. It is suggested that the stimulation of oxidative activity by Mn might be responsible for increasing the biotransformation of atrazine and for nonspecific transformations of other xenobiotic compounds.

Antioxidants↗

Response of spheroplasts and chelator-permeabilized cells of gram-negative bacteria to the action of the bacteriocins pediocin SJ-1 and nisin.

We have attempted to bypass the outer membrane (OM) barrier of Escherichia coli and Salmonella typhimurium with pediocin SJ-1 (as compared with nisin) using chelating agents as OM permeabilizers. EDTA, and to less extent EGTA, enabled nisin, but not pediocin SJ-1, to permeate the cell OM of E. coli, to have access to the cytoplasmic membrane and to cause subsequent permeability changes, indicated by an increase in ANS fluorescence intensity and a shift of its emission maximum. Such spectral changes did not occur when, prior to addition, EDTA was saturated with Ca2+ and Mg2+. ANS fluorescence data indicated that, in spite of the fact that pediocin SJ-1 did traverse the EDTA-permeabilized OM of E. coli, it did not cause perturbation of its cytoplasmic membrane and was, therefore, unable to cause cell death. Spheroplasts prepared from E. coli were lysed when treated with nisin but not with pediocin SJ-1. We suggest that the resistance of Gram-negative bacteria to pediocin SJ-1 is due not only to this material's inability to permeate the OM but also (in contrast to nisin) to its inability to interact with the cytoplasmic membrane.

Animals↗

Purification, partial characterization and plasmid-linkage of pediocin SJ-1, a bacteriocin produced by Pediococcus acidilactici.

Pediococcus acidilactici SJ-1, isolated from a naturally-fermented meat product, produced an antibacterial agent active against selected strains of Lactobacillus spp., Clostridium perfringens and Listeria monocytogenes. The agent was bactericidal against sensitive indicators, and sensitive to proteolytic enzymes; it was identified as a bacteriocin, and was designated as pediocin SJ-1. It was stable over a wide pH range (3-9), and apparently most stable in the lower part of that range. At pH 3.6, pediocin SJ-1 was stable at heat-processing temperatures within the range 65-121 degrees C; its activity decreased significantly, however, when it was heated at pH 7.0. The activity of pediocin SJ-1 on sensitive indicator cells was lost in the presence of alpha-amylase, suggesting that it contains a glyco moiety, necessary for its antibacterial action. Native pediocin SJ-1 exists in the form of monomers and aggregates (with molecular weights in the range 80-150 kDa). Pediocin SJ-1 was purified 262-fold by direct application of cell-free supernatant fluids to a cation-exchange chromatography column, and was resolved by SDS-PAGE as a single peptide band with a MW of ca 4 kDa. The original pediocin SJ-1-producing strain (bac+) harbours three plasmids of 4.6, 23.5, and 45.7 MDa. Production of pediocin SJ-1, but not immunity to SJ-1, is associated with the 4.6 MDa plasmid.

Bacteriocins↗

Isolation and Characterization of a Novel Atrazine Metabolite Produced by the Fungus Pleurotus pulmonarius, 2-Chloro-4-Ethylamino-6-(1-Hydroxyisopropyl)Amino-1,3,5-Triazine.

The white rot fungus Pleurotus pulmonarius exhibited metabolism of atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) in liquid culture, producing the dealkylated metabolites desethylatrazine, desisopropylatrazine, and desethyl-desisopropylatrazine. A fourth, unknown metabolite was also produced. It was isolated and was identified as 2-chloro-4-ethylamino-6-(1-hydroxyisopropyl)amino-1,3,5-triazine by gas chromatography-mass spectrometry, Fourier transformed infrared spectroscopy, and H nuclear magnetic resonance analysis. The structure of this metabolite was confirmed by chemical synthesis of the compound and comparison with the fungally produced metabolite.

Journal Article↗

Protection against tabun toxicity in mice by prophylaxis with an enzyme hydrolyzing organophosphate esters.

We demonstrate here the correlation between protection afforded by pretreatment alone with parathion hydrolase purified from Pseudomonas sp. against tabun toxicity in mice and the kinetic parameters which are assumed to determine the in vivo detoxification of tabun by the same enzyme. Results show that 15 and 22 micrograms of parathion hydrolase per animal conferred a protective ratio of 3.94 and 5.65 respectively, against tabun toxicity, without post-exposure treatment.

Animals↗

Accumulation of Scoparone in Heat-Treated Lemon Fruit Inoculated with Penicillium digitatum Sacc.

Phytoalexin scoparone (6,7-dimethoxycoumarin) generally was not detected in noninoculated lemon fruit (Citrus limon [L.] Burm., cv Eureka) but accumulated in fruit after inoculation with Penicillium digitatum Sacc. A much greater increase in the amount of scoparone was found in fruit exhibiting an incompatible response to Penicillium after heat treatment at 36 degrees C for 3 days. Heat treatment prevented development of decay in the inoculated fruit. The concentration of the compound after inoculation continued to increase during and after the heat treatment period, reaching 178 micrograms per gram fresh weight of the flavedo 6 days after the heat treatment. Changes in scoparone concentration in fruit were closely correlated with the changes in the antifungal activity of the fruit extract. A low concentration of the phytoalexin was detected in fruit injured mechanically. Scoparone also accumulated in the fruit following ultraviolet illumination; the concentration of the compound was dose-dependent. Median effective dose values of the inhibition of germ tube elongation and spore germination of P. digitatum were 29 and 46 micrograms per milliliter, respectively. Our findings suggest that the rapid increase in scoparone concentration plays an important role in the increased resistance of heat-treated lemon fruit to infection by P. digitatum.

Journal Article↗

Reduction of nitrosubstituted aromatic compounds by the halophilic anaerobic eubacteria Haloanaerobium praevalens and Sporohalobacter marismortui.

The moderately halophilic, obligately anaerobic eubacteria Haloanaerobium praevalens DSM 2228 and Sporohalobacter marismortui ATCC 35420 are able to reduce a variety of nitrosubstituted aromatic compounds at a high rate to the corresponding amines. Compounds degraded included nitrobenzene, o-nitrophenol, m-nitrophenol, p-nitrophenol, nitroanilines, 2,4-dinitrophenol, and 2,4-dinitroaniline. Most of these compounds, when added at concentrations of 50 to 100 mg/liter, were completely transformed within 24 h, but at the highest concentrations growth rates were somewhat lowered. Growth of H. praevalens in the presence of 14C-labeled p-nitrophenol showed that the compound was not incorporated by the cells or degraded to acid-volatile compounds.

Anaerobiosis↗

Effect of organic amendments on bacterial multiplication in lake water.

In Cayuga Lake water amended with 30 micrograms of glucose or amino acids per ml, an added strain of Pseudomonas fluorescens and indigenous bacteria grew extensively, Pseudomonas sp. B4 and two rhizobia multiplied at a moderate extent, and introduced Escherichia coli and Klebsiella pneumoniae multiplied but to only a slight degree. The amendments did not enhance growth of Micrococcus flavus and Arthrobacter citreus, and an asporogenous strain of Bacillus subtilis decreased in numbers. The pseudomonads, rhizobia, E. coli and K. pneumoniae multiplied extensively when inoculated into sterile lake water amended with amino acids, A. citreus grew to a slight extent, but the numbers of M. flavus and B. subtilis did not change appreciably. In nonsterile lake water amended with 30 micrograms of Trypticase soy broth per ml, the indigenous bacteria greatly increased in abundance, the pseudomonads, rhizobia, and E. coli developed to a lesser extent, the numbers of K. pneumoniae, A. citreus and M. flavus showed little increase, and B. subtilis decreased in numbers. Tests in pure culture containing 2 to 64 micrograms of Trypticase soy broth per ml demonstrated good growth of P. fluorescens, Pseudomonas sp. B4, and the rhizobia at all concentrations; an initial decline followed by growth of E. coli, K. pneumoniae and R. leguminosarum biovar phaseoli at low concentrations; little or no growth or decline at the low levels but multiplication at the high levels by A. citreus and B. subtilis; and decline of M. flavus. It is proposed that the apparent Ks value, mumax value, length of lag phase and resistance to stress can be used to predict behavior of bacteria in lake water receiving low levels of organic nutrients.

Amino Acids↗

Scanning Electron Microscope Studies of Interactions between Agaricus bisporus (Lang) Sing Hyphae and Bacteria in Casing Soil.

Relationships between the hyphae of Agaricus bisporus (Lang) Sing and bacteria from the mushroom bed casing layer were examined with a scanning electron microscope. Hyphae growing in the casing layer differed morphologically from compost-grown hyphae. Whereas the compost contained thin single hyphae surrounded by calcium oxalate crystals, the casing layer contained mainly wide hyphae or mycelial strands without crystals. The bacterial population in the hyphal environment consisted of several types, some attached to the hyphae with filamentlike structures. This attachment may be important in stimulation of pinhead initiation.

Journal Article↗

Aerotactic response of Azospirillum brasilense.

Five strains of Azospirillum brasilense and two of Azospirillum spp., from Israel, responded to self-created and preformed oxygen gradients by forming aerotactic bands in capillary tubes and actively moving toward a specific zone with low dissolved oxygen. Increasing the oxygen concentration in capillaries containing phosphate buffer increased the number of attracted bacteria and decreased band velocity. High O2 concentrations and H2O2 temporarily repulsed the bacteria, causing the formation of a bacterial arc around the capillary mouth. There was no band formation under anaerobic conditions, although the bacteria remained highly motile. Exogenous energy sources were unnecessary for aerotaxis in Azospirillum spp. The addition of oxidizable substrates to the capillary slightly enhanced aerotaxis, possibly by accelerating O2 consumption. Aerotactic band formation was affected by pH, bacterial concentration and age, incubation time, and respiratory inhibitors, but not by the lack of combined nitrogen in the growth medium. It is proposed that aerotaxis plays a role in the capacity of Azospirillum spp. to reach an environment suitable for N2 fixation.

Aerobiosis↗

Effects of Temperature, Nitrogen Fertilization, and Plant Age on Nitrogen Fixation by Setaria italica Inoculated with Azospirillum brasilense (strain cd).

The association between the nitrogen-fixing bacterium Azospirillum brasilense (strain cd) and the grass Setaria italica was studied under different environmental and soil conditions. Highest acetylene reduction rates in intact plants were observed at the booting stage of Setaria (2350 nmol ethylene produced hour(-1) plant(-1)) at 27 C. Higher temperatures, up to 32 C, enhanced ethylene reduction. Significant increases in shoot dry weight, panicle weight, and length were obtained in inoculated plants fertilized with suboptimal NH(4)NO(3) levels. The increase in nitrogen content of plants inoculated with A. brasilense was shown to be due to N(2) fixation. This was demonstrated by growing plants in washed quartz sand with no combined nitrogen. The bacteria also increased branching and development of roots. It was concluded that inoculation of Setaria with A. brasilense may lead both to increases in plant yield and saving of nitrogen fertilizer.

Journal Article↗

Citric Acid Fermentation by Aspergillus niger on Low Sugar Concentrations and Cotton Waste.

The possible use of cotton waste as a carbohydrate source of citric acid production by Aspergillus niger was examined. No citric acid was produced when A. niger was grown on cotton waste as a sole carbon source. In two-stage fermentations, however, mycelium obtained from surface cultures in cotton waste medium yielded more citric acid when transferred to sucrose-containing media than when directly inoculated to sucrose-containing media. It is concluded that cotton waste can be used for saving sucrose and for increasing yields of citric acid fermentation by A. niger.

Journal Article↗

Increase in Dry Weight and Total Nitrogen Content in Zea mays and Setaria italica Associated with Nitrogen-fixing Azospirillum spp.

The association between nitrogen-fixing bacteria from the genus Azospirillum and the grasses Zea mays and Setaria italica was investigated in sterilized Leonard-jar assemblies. Nitrogen-fixing bacteria isolated from Cynodon dactylon roots in Israel and Azospirillum brasilense (Sp-7, Sp-80, and Cd) were examined. C(2)H(2) reduction activity was detected in systems containing 0.0 to 0.08 but not in those containing 0.16 gram per liter NH(4)NO(3). The organisms tested significantly increased plant dry weight (50-100%), total N content of leaves (50-100%) and C(2)H(4) production (300-1000 nanomoles C(2)H(4) per plant per hour). Highest C(2)H(2) reduction activities were obtained above 30 C and with high light intensities. Significant increases in S. italica dry weight (DW) and nitrogen (N) content were observed in sand (DW = 80%, N = 150%), sandy loam soil (DW = 80%, N = 75%) and loess (DW = 37%, N = 25%). The results obtained in this work clearly demonstrate the potential benefit of inoculating grasses with Azospirillum.

Journal Article↗

An increase in nitrogen content of Setaria italica and Zea mays inoculated with Azospirillum.

Bacteria belonging to the genus Azospirillum isolated from Cynodon dactylon roots in Israel were compared with Azospirillum brasilense from Brazil and California for their ability to fix nitrogen in association with grasses under greenhouse conditions. The plants were grown in a system which avoided cross inoculation from the inoculated soil to the control, while maintaining the natural soil microflora and humidity level in the soil close to field capacity. The organisms tested significantly increased the dry weight of Zea mays and Setaria italica leaves, the total nitrogen content of these leaves (as measured by the Kjeldahl method), and supported acetylene reduction in intact nonsterile systems as compared with the noninoculated controls. Ethylene production in intact systems could be detected after 6h and was linear for 72h, providing a constant soil temperature (28-32 degrees C) was maintained.

Acetylene↗

Comparative studies of nitrogen-fixing bacteria associated with grasses in Israel with Azospirillum brasilense.

Free-living, dinitrogen-fixing bacteria associated with roots of grasses were isolated from several locations in Israel. Bacteria with characteristics similar to those of Azospirillum were isolated from Cynodon dactylon roots and were compared with Azospirillum brasilense from Brasil (Sp-7) and from California (Cd). Colonies of the Israeli isolates were yellow and consisted of curved rods, 0.5-0.6 micron in diameter with polar flagella, whereas colonies of A. brasilense were pink (Sp-7) and red (Cd) and the cells were 1.0-1.1 micron in diameter with polar flagella. Ultraviolet absorption spectra of soluble c and membrane-bound c and b cytochromes were similar in all isolates. When grown in semisolid agar medium with or without ammonium chloride all isolates formed a growth zone below the surface. However, they grew best under aerobic conditions in liquid medium containing NH4Cl. All isolates could use salts of malate and lactate, arabinose, and galactose, but not mannitol, as sole carbon sources; they did not need biotin to shorten their lag phase. One Israeli isolate was capable of growing and fixing nitrogen with glucose as a sole carbon source. The Israeli isolates formed aggregates above pH 7.6 in liquid or semisolid medium and were capable of reducing nitrate to nitrogen gas under anaerobic conditions.

Bacteria↗

Investigations on the photosynthetic sulfur bacterium Chlorobium phaeobacteroides causing seasonal blooms in Lake Kinneret.

Between May and December, the annual stratification period in Lake Kinneret, sulfide is formed and accumulates in the hypolimnion. In July-August a large population (up to 10(6) cells/mL) of green, photosynthetic, sulfur bacteria develops at the boundary of the oxidative and reductive zones of the water column lasting for 3--8 weeks. These bacteria were isolated from the lake and identified as Chlorobium phaeobacteroides. Optimal growth conditions included 160 mg S=L-1 and light intensities of 5--30 micron Einstein (micron E) m-2s-1. Glucose and acetate augmented growth when added to the mineral medium. The lowest light intensity which still supported growth was 0.3 micron E m-2s-1 when acetate was present and 1.0 micron E m-2s-1 when no organic substrate was present. Under complete darkness, either with or without organic substrate, the bacteria die. Photosynthetic activity was higher when no organic compound was added to the medium. Uptake of acetate was light-dependent. In the lake the photosynthetic activity of the bacteria is low because of the limited light intensity (0.3 micron E m-2s-1) at the bloom layer. It is suggested that the appearance and the disappearance of the bloom are caused by the influence of the daily internal seiche.

Acetates↗