Effects of some tricyclic psychopharmacons and structurally related compounds on motility of Proteus vulgaris.

A simple test for the evaluation of drugs interfering with bacterial motility was established with Proteus vulgaris. With this model, promethazine, 7-hydroxy-chlorpromazine, imipramine, 7,8-dioxochlorpromazine and acridine orange were shown to exert significant motility and swarming inhibitory action on Proteus vulgaris strains at subinhibitory concentrations. Quinidine enhanced the antimotility effect of promethazine. The antimotility effect of promethazine was synergized by proton pump inhibitors omeprazole and abscissic acid, but antagonized by extracellular potassium and sodium ions.     

Antifeedant effects of azadirachtin and structurally related compounds on lepidopterous larvae

The antifeedant activity of azadirachtin, azadirachtin-derivatives and related limonoids was assessed in choice and no-choice bioassays against four species of Lepidoptera: Spodoptera littoralis, Spodoptera frugiperda, Heliothis virescens and Heliothis armigera. The choice bioassay showed that the feeding behaviour of S. littoralis was affected by more of the compounds than that of either S. frugiperda or H. virescens. H. armigera was the least affected. Azadirachtin and dihydroazadirachtin were the most potent of the 40 compounds tested. The results showed that hydrogenation of the C-22,23 double bond did not decrease antifeedant activity and the nature of the substitutes at C-1, C-3 and C-11 were important. Molecules with bulky substitutes at either C-22 or C-23 were usually ineffective antifeedants as were compounds lacking an epoxide. Compounds recorded as active antifeedants in the choice bioassay were not always as active in the no-choice test. The value of the bioassays in assessing the mode of action of the compounds is discussed.

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Résumé L'activité phagodissuadante de l'azadirachtine, de ses dérivés et des limonoïdes voisins sur 4 espèces de lépidoptères: Spodoptera littoralis, S. frugiperda, Heliothis virescens et H. armigera a été évaluée par des expériences avec et sans choix. Les expériences de choix ont montré que le comportement alimentaire de S. littoralis était modifié par plus de substances que celui de S. frugiperda ou H. virescens. Celui de H. armigera était le moins modifié. Les 2 substances les plus puissantes parmi les 40 examinées, ont été l'azadirachtine et le dihydroazadirachtine. Ces résultats montrent que l'hydrogénation de la double liaison C-22,23 ne réduit pas l'activité phagodissuadante et que la nature des substitutions en C-1, C-3 et C-11 sont importantes. Les molécules avec des substitutions volumineuses en C-22 ou C-23 sont généralement des phagodissuadants aussi inefficaces que ceux ayant perdu un époxide. Les substances notées comme phagodissuadants actifs dans les expériences de choix ne sont pas toujours aussi actives en absence de choix. La valeur des tests dans l'évaluation du mode d'action des substances est discuté.

     

Effects of phenolic compounds on growth and metabolic activities of Chlorella vulgaris and Scenedesmus bijugatus isolated from soil

Phenol and three nitrophenols (o-nitrophenol, m-nitrophenol, p-nitrophenol), commonly occurring pollutants in natural eco-systems, were tested for their toxic effects on soil isolates of Chlorella vulgaris and Scenedesmus bijugatus, growing under phototrophic, photoheterotrophic and heterotrophic conditions. The toxicity criteria included cell number, chlorophyll, total protein and carbohydrate content, ¹⁴CO₂ uptake and in vivo nitrate reductase activity. Both C. vulgaris and S. bijugatus were sensitive to the pollutants when the cultures were grown under phototrophic or heterotrophic conditions. However, the toxicity was found reversed or alleviated upon photoheterotrophic growth of the cultures. Transmission electron microscope studies revealed various cytological abnormalities in C. vulgaris in the presence of phenolics at algistatic levels.     

Site-directed mutagenesis of cysteine to threonine in Proteus vulgaris urease active site increases enzyme activity and stability

Cysteine-319 belongs to the flexible flap at the active site of Proteus vulgaris urease. Replacing this cysteine by threonine resulted in a 20-fold increase of specific activity. Temperature stability increased, susceptibility to inhibition by dipyridyl disulfide decreased, and pH optimum shifted from 8 to 6.9. K _m (35 to 12 mM) and V_max (47.4 to 1.8 mol min^–1) were substancially altered. Both variants of the enzyme were irreversibly inhibited by phenylmethanesulfonyl fluoride.     

The toxicity of phenolic compounds to soil algal population and toChlorella vulgaris andNostoc linckia

Summary p-Nitrophenol (PNP),m-nitrophenol (MNP), 2,4-dinitrophenol (DNP) and catechol were tested for their effects on algal population in a soil and on pure cultures of two algae isolated from soil. Both PNP and MNP, even at 0.5 kg ha⁻¹ level were toxic to the soil algae; high doses effected increase in toxicity. Inhibition of algae was relatively more with PNP compared to the other two nitrophenols. Catechol treatment up to 1.0 kg ha⁻¹ led to a significant initial enhancement of algae with a subsequent far less toxic effect. The toxicity of the phenolic compounds towardChlorella vulgaris, a green alga andNostoc linckia, a blue-green alga, decreased in the order: MNP≧PNP>DNP>Catechol. However, algicidal or algistatic effect of the test chemicals was fairly more againstC. vulgaris, suggesting that the eukaryotic alga is highly sensitive to such soil pollutants compared to the prokaryotic alga.     

Effects of ethylene and inhibitors of ethylene synthesis and action on nodulation in common bean (Phaseolus vulgaris L.)

The ethylene releasing compound, 2-chloroethylphosphonic acid (ethephon) inhibited nodule development in common bean (Phaseolus vulgaris L.) plants. In contrast, inhibitors of ethylene synthesis or its physiological activity enhanced nodulation. In a co-culture of bean seeds and rhizobia, ethephon inhibited rhizobial growth while inhibitors of ethylene synthesis or action did not influence the growth and proliferation of rhizobia. These data emphasize the role of ethylene as a regulator of nodulation in determinate nodulators and indicate that the ethylene signaling pathway involved in the nodulation process is not limited to the plant host but also involves the bacterial symbiont.     

Intra-specific variation in the inhibitory effects of Thymus vulgaris (Labiatae) monoterpenes on seed germination

The ecological role of plant secondary compounds has received a great deal of attention yet little is known regarding variation in the ecological significance of different compounds produced by a single species. This is particularly pertinent to species where variation in compound presence is under genetic control. In this study we have quantified variation in the inhibitory effects of the six dominant monoterpene oils produced by different genotypes (or chemotypes) of Thymus vulgaris on achene germination of an associated species (Brachypodium phoenicoides), and seeds of T. vulgaris itself, in controlled experimental conditions. The experiments involved the germination of seeds of the two species with crushed leaves or pure essences of the six chemotypes, i.e. four experimental trials. A significant inhibitory effect against B. phoenicoides was found for both crushed leaves and the range of concentrations of pure oils used in the trial. The two phenolic compounds had a greater effect than the non-phenolic leaves and pure oils. The significant differences between non-phenols and the controls declined faster over time for crushed leaves and low concentrations of the pure essences than did that between the phenolics and the controls. There was a significant inhibitory effect against the germination of T. vulgaris seeds for the crushed leaves and the pure oils, although the contrast between the non-phenols and the phenols was not significant. The effect of the different oils varied depending on the seed source of the thyme seeds used in the trial. This is suggested to be due to differences in the rates of outcrossing among the sampled populations. The effects of the oils on the associated species, B. phoenicoides, were not found to be greater than their effects on the germination of thyme seeds. The results are discussed in the context of a potential role for such inhibitory effects in relation to the onset of rainfall and suitable germination conditions.     

Cell surface properties as factors involved in Proteus vulgaris adhesion to stainless steel under starvation conditions

The purpose of these investigations was to evaluate the influence of limited nutrient availability in the culture medium on Proteus vulgaris biofilm formation on surfaces of stainless steel. The relationship between the P. vulgaris adhesion to the abiotic surfaces, the cellular ATP levels, cell surface hydrophobicity and changes in the profiles of extracellular proteins and lipopolysaccharides was examined. In all experimental variants the starvation conditions induced the bacterial cells to adhere to the surfaces of stainless steel. Higher ATP content and lower cell surface hydrophobicity of P. vulgaris cells was observed upon nutrient-limited conditions. Under starvation conditions a reduction in the levels of extracellular low molecular weight proteins was noticed. High molecular weight proteins formed the conditioning layer on stainless steel plates, making the bacteria adhesion process more favorable. The production of low molecular weight carbohydrates promoted more advanced stages of P. vulgaris biofilm formation process on the surfaces of stainless steel upon starvation.     

Tryptophan indole-lyase from Proteus vulgaris: kinetic and spectral properties

An efficient method for purification of recombinant tryptophanase from Proteus vulgaris was developed. Catalytic properties of the enzyme in reactions with L-tryptophan and some other substrates as well as competitive inhibition by various amino acids in the reaction with S-o-nitrophenyl-L-cysteine were studied. Absorption and circular dichroism spectra of holotryptophanase and its complexes with characteristic inhibitors modeling the structure of the principal reaction intermediates were examined. Kinetic and spectral properties of two tryptophanases which markedly differ in their primary structures are compared. It was found that although the spectral properties of the holoenzymes and their complexes with amino acid inhibitors are different, the principal kinetic properties of the enzymes from Proteus vulgaris and Escherichia coli are analogous. This indicates structural similarity of their active sites.     

Effects of cooling and freezing on the motility of Ostrea edulis (L., 1758) spermatozoa after thawing

The aim of this work was to evaluate the effects of temperature, cryoprotectant agents and freezing curves on sperm motility of Ostrea edulis. All phases of cryopreservation were studied (evaluation of semen motility pattern, choice of cryoprotectants and freezing rates) to restore after thawing the motility characteristics distinctive of fresh semen.To assess the temperature effects on sperm motility, semen was activated using four different temperatures (25, 18, 10 and 3 °C). Sperm aliquots were maintained inactive at these temperatures for 1 and 3 h, then activated with FSW at same temperature of conservation. Sperm was activated and incubated to 3 °C with dimethylsulfoxide (Me₂SO), ethylene glycol (EG), 1–2 propylene glycol (PG) (5%, 7%, 10% and 15% final concentrations), glycerol (GlOH; 5%, 10% and 15% final concentrations) and methanol (MetOH; 4% and 10% final concentrations) for 10, 20 and 30 min. A first evaluation of freezing rates was made by testing four freezing curves: −1, −3, −6 and −10 °C/min. Then, an optimization was made by testing four freezing curves: −2.5, −3.0, −3.5 and −4 °C/min.The selected temperature for short term conservation has been 3 °C, because only this temperature has allowed good sperm motility conservation after 3 h of dry-storage; this is a time sufficient to conduct cryopreservation procedures. The sperm showed a particular sensitivity to GlOH and PG to all tested concentrations and to 15% Me₂SO. EG and MetOH to all concentrations and Me₂SO to concentrations lower than 15% have not shown significant toxic effects. The freezing rate −3 °C/min using 15% EG has shown an highest percentage of RVF (rapid, vigorous and forward) spermatozoa (class 3, about 75% of fresh semen) and an highest sperm motility duration.     

肽酶PepT参与副溶血弧菌的浮游运动和生物被膜形成

pepT基因编码一种金属依赖性肽酶T (peptidase T,PepT),能特异性催化三肽N端氨基酸,因此也称为氨肽酶T。研究发现大多数氨肽酶参与细菌蛋白质新陈代谢和调节三肽活性,但关于PepT在细菌毒力及致病性方面的报道较少。[目的]本文选取PepT为研究对象,研究其对副溶血弧菌生物学特性及致病性的影响。[方法]通过构建缺失株ΔpepT和回补株CΔpepT,比较菌株在运动性、生物被膜、环境耐受、细胞毒性等方面的差异。[结果]与野生株相比,ΔpepT缺失株的极性鞭毛转录水平极显著下降,浮游运动能力降低;同时生物被膜形成能力减弱,而细菌群集运动及环境耐受能力无显著差异。此外,缺失pepT基因会导致副溶血弧菌的细胞毒性和小鼠毒力作用显著下降。[结论]pepT基因与副溶血弧菌浮游运动和生物被膜形成能力相关,并且影响其致病性。     

No direct linkage between proton pumping and photosynthate unloading from seed coats of Phaseolus vulgaris L.

The energization of the active sucrose release from bean seed-coat halves was investigated. For this purpose, seed coat tissues adjacent to the apoplastic space were exposed to a variety of treatments and proton and photosynthate release were measured. Fusicoccin (10^–5 moll^–1) stimulated proton pump activities. Orthovanadate (2×10^–4 moll^–1) and abscisic acid (10^–5 moll^–1) diminished the proton extrusion evoked by fusicoccin. Fusicoccin inhibited sucrose release, whereas orthovanadate and abscisic acid stimulated it. Addition of 100 mmoll^–1 K⁺ had a promotory effect on photosynthate unloading, fading away with time. This extra unloading was linearly related to an enhanced proton loss. It was concluded that the photosynthate unloading apparently is not a proton/sucrose antiport and that a pump-leak system for photosynthate release is unlikely. A tentative model for photosynthate/proton symport not directly linked to proton pumping is presented as the mechanism of unloading.Abbreviations ABA abscisic acid - CCCP carbonyl cyanide m-chlorophenyl hydrazone - DTE diethioerythritol - FC fusicoccin - MES 2-(N-morpholino) ethanesulfonic acid monohydrate - NEM n-ethylmaleimide - PCMBS p-chloromercuriphenylsulfonic acid - TRIS 2-amino-2-(hydroxymethyl) propane-1,3 diol - VAN sodium orthovanadate     

The spatial distribution of cations in nematocytes of Hydra vulgaris

The spatial distribution of cations was assayed qualitatively and quantitatively in tentacular nematocytes of Hydra vulgaris in a scanning transmission electron microscope by means of x-ray microanalysis performed on 100 nm thick freeze-dried cryosections. The matrix of undischarged cysts (stenoteles, desmonemes and isorhizas) was found to contain mainly K⁺. In isolated nematocysts of Hydra the intracapsular potassium can be readily substituted by practically any other mono- and divalent cation (Na⁺, NH₄ ⁺, Mn²⁺, Co²⁺, Mg²⁺, Ca²⁺, Fe²⁺) all, except Fe²⁺, without impairing the ability of the cyst to respond to the chemical triggering with dithioerythritol or proteases. Monovalent cations increase the osmotically generated intracapsular pressure compared to divalent ions.     

The effect of copper toxicity on the contents of nitrogen compounds in Silene vulgaris (Moench) Garcke

The effect of copper on the uptake of nitrogen and the tissue contents of inorganic nitrogen, amino acids and proteins were studied in cooper-sensitive Silene vulgaris (Moench) Garcke, grown at different nitrogen sources (NH₄ ⁺ or NO₃ ^-). All the toxic copper levels tested, i.e. 4, 8, 16 M Cu²⁺, strongly inhibited the uptake of nitrogen, especially of NO₃ ^-, and decreased the content of NO₃ ^-, amino acids and proteins. Especially at 4 and 8 M Cu²⁺, NH₄ ⁺ accumulated in the plants, suggesting that the conversion of NH₄ ^- into amino acids was inhibited.     

Production of volatile aroma compounds by bacterial strains isolated from different surface-ripened French cheeses

Twelve bacterial strains belonging to eight taxonomic groups: Brevibacterium linens, Microbacterium foliorum, Arthrobacter arilaitensis, Staphylococcus cohnii, Staphylococcus equorum, Brachybacterium sp., Proteus vulgaris and Psychrobacter sp., isolated from different surface-ripened French cheeses, were investigated for their abilities to generate volatile aroma compounds. Out of 104 volatile compounds, 54 volatile compounds (identified using dynamic headspace technique coupled with gas chromatography-mass spectrometry [GC-MS]) appeared to be produced by the different bacteria on a casamino acid medium. Four out of eight species used in this study: B. linens, M. foliorum, P. vulgaris and Psychrobacter sp. showed a high flavouring potential. Among these four bacterial species, P. vulgaris had the greatest capacity to produce not only the widest varieties but also the highest quantities of volatile compounds having low olfactive thresholds such as sulphur compounds. Branched aldehydes, alcohols and esters were produced in large amounts by P. vulgaris and Psychrobacter sp. showing their capacity to breakdown the branched amino acids. This investigation shows that some common but rarely mentioned bacteria present on the surface of ripened cheeses could play a major role in cheese flavour formation and could be used to produce cheese flavours.     

Activity of salicylic acid on the growth and biochemism of Chlorella vulgaris Beijerinck

This study was conducted to investigate the influence of salicylic acid (SA) on the growth and changes of nucleic acids, protein, photosynthetic pigments, sugar content and photosynthesis levels in the green alga Chlorella vulgaris Beijerinck (Chlorophyceae). The most significant changes in the content of nucleic acids and proteins was observed at the concentration 10⁻⁴ M SA between 8 and 12 day of cultivation. This concentration of SA increased the number of cells (about 40 %) and content of proteins (about 60 %) and its secretion to the medium. The slight stimulation of protein secretion occurred on the 12th day of cultivation at concentration 10⁻⁴ M, while in the range of 10⁻⁵ M to 10⁻⁶ M the protein secretion was inhibited. SA also stimulated the content of nucleic acids, especially RNA by 20–60 %, compared with the control. The most stimulating influence upon the contents of chlorophylls a and b (50–70 %), total carotenoids (25–57 %), sugar (27–41 %) and intensity of net photosynthesis (18–33 %) was found at 10⁻⁴ M of SA. At the concentration of 10⁻⁶ M SA the slight inhibition of growth and biochemical activity of the algae was recorded at the first days of cultivation.     

Alternaria brasiliensis sp. nov., a leaf pathogen on Phaseolus vulgaris

Alternaria brasiliensis sp. nov., was isolated from leaves of common bean, Phaseolus vulgaris L., showing punctiform non concentric leaf spots of brown color. Besides the symptomatological differences, the new Alternaria species presents distinct type of arrangement of conidia chain, body and beak size. The disease was observed in Montanha county, State of Espirito Santo, Brazil, but not yet reported in the literature. The causal agent of the disease is now described by the first time. The epithet used here is referring to the country where the species has been found. This revised version was published online in June 2006 with corrections to the Cover Date.     

Studies on the incorporation of CO2 into starch by Chlorella vulgaris

Chlorella vulgaris was grown photosynthetically in batch culture under nitrogen sufficiency or nitrogen limitation. The starch content of the cells was measured as the amount of glucose released by enzymic hydrolysis of partially purified starch. Nitrogen sufficient algae contained approximately 20% of their dry weight as starch, whereas in nitrogen limited cells starch comprised up to 55% of the cellular dry weight. Starch production was pH dependent; optimal production of starch was achieved between pH 7.5 and 8.0. Optimal growth of C. vulgaris occurred at pH 7.0. Carbon yield experiments showed that for every gram of carbon consumed 0.5 g of starch (glucose) could be recovered. author for correspondence     

Ferredoxin and ferredoxin-NADP-oxidoreductase in leaves ofPhaseolus vulgaris L.

During light-induced greening of 10-dayold etiolated bean seedlings a strong increase is observed of ferredoxin (Fd) and of ferredoxin-NADP-oxidoreductase (FNR; E.C. 1.6.99.4) activity, both known to reside in chloroplasts. The production of Fd starts immediately upon illumination and proceeds almost linearly for at least the next 72 h. It is inhibited by chloramphenicol (CAP) but not by cycloheximide (CHI), beit that in the presence of the latter Fd synthesis was impaired after 48 h of illumination. We conclude that for the elaboration of functional Fd in greening chloroplasts protein synthesis on chloroplast ribosomes is required. The increase of FNR activity shows a lag of about 24 h and is blocked by both CAP and CHI. When CAP is applied at 24 h after the illumination started it has no effect. We suggest that the synthesis of FNR on cytoplasmic ribosomes requires prior synthesis of protein(s) on chloroplast ribosomes.The nature of possible interactions between chloroplasts and cytoplasm is discussed.Abbreviations CAP D-threo-chloramphenicol - CHI cycloheximide - DCIP dichlorophenol-indophenol - DEAE diethylaminoethyl - Fd Ferredoxin - FNR ferredoxin-NADP-oxidoreductase - NAR nitrate reductase - NIR nitrite reductase