A copper-containing protein that inhibits nitrite reductase from Pseudomonas aeruginosa

A non-blue copper-containing glycoprotein was isolated from Pseudomonas aeruginosa. The protein has a molecular mass of 10 kDa and contains 1 atom of EPR-detectable type II copper. The protein inhibits oxidation of both azurin and cytochrome c-551 catalyzed by nitrite reductase from Ps. aeruginosa. Thus, it may be considered as an endogenous inhibitor of nitrite reductase.     

Molecular cloning of cis-acting regulatory alleles of the Bacillus subtilis amyR region by using gene conversion transformation.

Three cis-acting alleles (gra-10, gra-5, and amyR2) of the Bacillus subtilis amyR promoter locus each cause catabolite repression-resistance of amyE-encoded alpha-amylase synthesis. The gra-10, gra-5, and amyR2 alleles were transferred from the chromosomes of their respective hosts to a plasmid carrying the amyR1-amyE+ gene by the process of gene conversion which is carried out during transformation of competent B. subtilis by plasmid clones carrying homologous DNA. The cloned amyR promoter regions containing the gra-10 and gra-5 mutations were shown to confer catabolite repression-resistance in cis to the synthesis of chloramphenicol acetyltransferase encoded by the cat-86 indicator gene when subcloned into the promoter-probe plasmid pPL603B. Implications concerning both the regulation of amyR utilization and the process of gene conversion in B. subtilis are discussed.     

Colour-blind drivers' perception of traffic signals.

There is a dangerous and widespread assumption that colour-blind drivers drive as safely as drivers with normal vision. Colour-blind drivers have difficulties recognizing traffic and vehicle signals. However, simple and practical solutions such as changes to the design, colour and shape of signals will aid these drivers.     

Widespread cutaneous candidiasis and tinea infection masking mycosis fungoides

A 71-year-old female with a widespread double mycotic infection caused byC. Albicans andT. rubrum was discovered to be suffering from mycosis fungoides. Clinically she was found to have large, polycyclic erythematous plaques with scaly, slightly infiltrated borders, covering almost all areas of the glabrous skin, and also involving the scalp (with no hair penetration), the soles and palms, toe-webs, finger and toe nails; there was also perleche and oral thrush. Cultures yieldedC. albicans from most of the skin lesions, from the scalp, mouth, finger nails and urine and stool specimens, andT. rubrum from intermingled skin specimens, from the palms and soles and toe-nails. Blood culture was negative as were intracutaneous tests with fungal antigens and tuberculin. Histological examination confirmed the fungal invasion of the horny layer and at the same time revealed an underlying pathologic picture of mycosis fungoides, the lesions having been masked by the mycotic eruption. Intensive cytostatic and antifungal therapy led to a transient improvement but shortly thereafter there was a relapse of the fungal and lymphoproliferative manifestations and the patient died in septic shock.

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Résumé Une femme, agée 71 ans, ayant une double infection mycosique due àC. albicans et auT. rubrum, a été décélée qu'elle souffrait aussi d'un mycosis fungoïde. Au point de vue clinique elle avait de larges plaques érythémato-squameuses, polycicliques, aux bords légèrement infiltrés, répandues sur une grande partie de la peau glabre, paumes et plantes inclus, et affectant aussi le cuir chevelu (sans pénétration des poils), les ongles des doigts et des orteils. Les cultures ont permis d'isoler leC. albicans à partir de la majorité des lésions de la peau et du cuir chevelu, des ongles des doigts, ainsi que de la bouche, de l'urine et des selles. On a trouvé aussiT. rubrum dans des lésions cutanées entremêlées aux precedentes, dans la région palmaire et plantaire et dans les ongles des orteils. Les testes intradermiques aux antigens fongiques et à tuberculine ont été négatifs, ainsi que les hémocultures. L'examen histologique a montré l'invasion de la couche cornée par les champignons et en même temps a découvert un tableau pathologique sous-jacent d'un mycosis fungoïde, ayant été masqué par l'éruption fongique. Un traitement intensif par des produits cytostatiques et antifongiques a mené à une amélioration temporaire, mais brièvement après on a assisté à une réchute rapide des manifestations lymphoprolifératives et fongiques et la malade a décédé à la suite d'un état septique.

     

Identifying regions of membrane proteins in contact with phospholipid head groups: covalent attachment of a new class of aldehyde lipid labels to cytochrome c oxidase

A series of amine-specific reagents based on the benzaldehyde reactive group have been synthesized, characterized, and used to study beef heart cytochrome c oxidase reconstituted in phospholipid bilayers. The series contained three classes of reagents: lipid-soluble phosphodiesters having a single hydrocarbon chain, phospholipid analogues, and a water-soluble benzaldehyde. All reagents were either radiolabeled or spin-labeled or both. The Schiff bases formed by these benzaldehydes with amines were found to be reversible until the addition of the reducing agent sodium cyanoborohydride, whereas attachment of lipid-derived aliphatic aldehydes was not readily reversible in the absence of the reducing agent. The benzaldehyde group provides a convenient method of controlling and delaying permanent attachment to integral membrane proteins until after the reconstitution steps. This ensures that the lipid analogues are located properly to identify amine groups at the lipid-protein interface rather than reacting indiscriminately with amines of the hydrophilic domains of the protein. The benzaldehyde lipid labels attach to cytochrome c oxidase with high efficiency. Typically, 20% of the amount of lipid label present was covalently attached to the protein, and the number of moles of label incorporated per mole of protein ranged from 1 to 6, depending on the molar ratios of label, lipid, and protein. The efficiency of labeling by the water-soluble benzaldehyde was much less than that observed for any of the lipid labels because of dilution effects, but equivalent levels of incorporation were achieved by increasing the label concentration. Electron spin resonance spectra of a nitroxide-containing phospholipid analogue covalently attached to reconstituted cytochrome c oxidase exhibited a large motion-restricted component, which is characteristic of spin-labeled lipids in contact with the hydrophobic surfaces of membrane proteins. The line shape and splittings were similar for covalently attached label and label free to diffuse and contact the protein molecules in the bilayer, providing independent evidence that the coupling occurs at the protein-lipid interface. The distribution of the benzaldehyde reagents attached to the polypeptide components of cytochrome c oxidase was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The labeling pattern observed for the lipid analogues was not affected by the presence of the nitroxide moiety on the acyl chains but was dependent on the molar ratio of labeling reagent to protein.(ABSTRACT TRUNCATED AT 400 WORDS)     

Mechanism of the suppression of the spinal pain syndrome by serotonin derivatives

The ability of serotonin derivatives to stimulate cAMP accumulation in isolated nerve terminals and lumbar enlargement of the spinal cord of normal rats was compared. The effect of the compounds on the intensity of spinal pain syndrome was also assessed. It has been established that substitutes injected into NH2-group of serotonin in 5-OH position attenuate the ability to stimulate cAMP accumulation in synaptosomes, with the effect more pronounced with substitutes of larger volume. A certain correlation between the ability of serotonin derivatives to stimulate adenylate cyclase in vivo and in vitro, on the one hand, and their analgetic effect, on the other hand, is suggested.