Multiple native flavin reductases in camphor-metabolizing Pseudomonas putida NCIMB 10007: functional interaction with two-component diketocamphane monooxygenase isoenzymes.

Although they have been studied for nearly 50 years, the source of the FMNH2 needed for effective biooxidation by the 2,5- and 3,6-diketocamphane monooxygenase (DKCMO) isoenzymes induced by the growth of Pseudomonas putida NCIMB 10007 (ATCC 17453) on camphor remains incompletely characterized. Prior studies have focussed exclusively on enzymes present in cells harvested during late-exponential-phase growth despite considerable circumstantial evidence that the flavin reductase (FR) component of these multicomponent monooxygenases is subject to growth-phase-dependent variation. In this study, a number of alternative FMNH2-generating activities, including both conventional FRs and enzymes also able to serve as ferric reductases, were isolated from camphor-grown cells, and the relative level, and hence potential contribution, of these various proteins shown to vary considerably depending on the point of harvest of NCIMB 10007 within exponential-phase growth.

Carbohydrate and Amino Acid Fermentation in the Free-Living Primitive Protozoon Hexamita sp.

Hexamita sp. is an amitochondriate free-living diplomonad which inhabits O(2)-limited environments, such as the deep waters and sediments of lakes and marine basins. C nuclear magnetic resonance spectroscopy reveals ethanol, lactate, acetate, and alanine as products of glucose fermentation under microaerobic conditions (23 to 34 muM O(2)).

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1. Aquatic bryophytes were sampled from 108 streams spanning over 3000 m of altitude in four regions of Nepal. Richness, cover and community composition were related to physicochemistry using multiple regression, DECORANA ordination and TWINSPAN.

Acoustic rhinometry: an explanation of some common artefacts associated with nasal decongestion.

The nasal cavities of 51 healthy volunteers were examined using acoustic rhinometry before and after nasal decongestant. Several specific dimensions were studied, which included the minimum cross-sectional area, and three volumes corresponding to the anterior, middle and posterior regions of the nasal airway. An average acoustic rhinometry trace was constructed for the whole group of subjects, before and after decongestion, from data extracted from the raw data files written to the computer hard disk for each subject.

Collagen orientation and molecular spacing during creep and stress-relaxation in soft connective tissues.

Collagen fibres form cross-helical, cross-ply or quasi-random feltworks in extensible connective tissues; strain-induced reorientation of these networks gives rise to the non-linear mechanical properties of connective tissue at finite strains. Such tissues are also generally viscoelastic (i.e.

Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA.

We report the design and evaluation of PCR primers 63f and 1387r for amplification of 16S rRNA genes from bacteria. Their specificity and efficacy were tested systematically with a bacterial species and environmental samples. They were found to be more useful for 16S rRNA gene amplification in ecological and systematic studies than PCR amplimers that are currently more generally used.

Pharmacological importance of stereochemical resolution of enantiomeric drugs.

Drug enantiomers have identical properties in an achiral environment, but should be considered as different chemical compounds. This is because they often differ considerably in potency, pharmacological activity and pharmacokinetic profile, since the modules with which they interact in biological systems are also optically active. Within biological systems, the metabolism of one isomer may be via a different pathway or occur at a different rate from that of the other isomer.

Oxygen uptake and antioxidant responses of the free-living diplomonad Hexamita sp.

The free-living anaerobic flagellate Hexamita sp. was observed to actively consume O2 with a K(m) O2 of 13 microM. Oxygen consumption increased linearly with O2 tension up to a threshold level of 100 microM, above which it was inhibited.

Acridine orange as an indicator of bacterial susceptibility to gentamicin.

We have studied the response of Escherichia coli NCTC10418 to gentamicin with flow cytometry. The susceptibility of individual bacterial cells to the antibiotic was assessed by differential staining using the metachromatic dye, acridine orange. Exponential phase cultures were exposed to the minimum bactericidal concentration of gentamicin and analysed at regular intervals over 90 min.

A flow cytometric study of antibiotic-induced damage and evaluation as a rapid antibiotic susceptibility test for methicillin-resistant Staphylococcus aureus.

Flow cytometry using the anionic membrane potential-sensitive fluorescent probe, bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)), enabled assessment of antibiotic-induced membrane perturbation in five clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and two antibiotic-sensitive reference strains, NCTC 6571 and 8325-4, after establishment of steady-state growth in liquid cultures inoculated from single colonies. Flow cytometric indications of the enhanced DiBAC4(3) uptake after treatment with vancomycin at 0.1, 1, 4 and 10 x MIC showed excellent comparison with viability losses quantified as cfu on solid agar in MRSA isolate QC.

The effects of agar concentration on the growth and morphology of submerged colonies of motile and non-motile bacteria.

The growth and morphology of submerged bacterial colonies was investigated. Five separate colonial forms were recognized depending both on species and on agar concentration. These were (i) branched, dendritic structures seen only with Bacillus cereus; (ii) lenticular colonies for all other species at high agar concentrations; (iii) small lobed to spherical colonies for non-motile organisms at low agar concentrations; (iv) and (v) large diffuse spherical colonies which can be further subdivided into 'snowball' or 'wispy' types for motile bacteria growing at agar concentrations below about 0.

The effects of adherence to silicone surfaces on antibiotic susceptibility in Staphylococcus aureus.

Sensitivity of Staphylococcus aureus to the antibiotics tetracycline, benzylpenicillin and vancomycin was found to decrease by 2-10 fold when cells were grown adherent to silicone catheter surfaces. Sensitivity to rifampicin and fusidic acid was not significantly altered in adherent cells. Susceptibility further decreased with increased adherence time prior to antibiotic challenge.

Interaction between piloting and beacon homing by rats in a swimming pool.

In three experiments, rats in a swimming pool were trained to find a submerged platform with a beacon attached to it. For some rats this beacon unambiguously identified the location of the platform; for others the beacon was made ambiguous by placement of an identical beacon in a different part of the pool. Test trials, in the absence of the platform and the beacons, revealed more persistent searching near the original location of the platform if the beacon attached to the platform had been ambiguous.

Cloning and characterization of two glutathione S-transferases from a DDT-resistant strain of Anopheles gambiae.

Two cDNA species, aggst1-5 and aggst1-6, comprising the entire coding region of two distinct glutathione S-transferases (GSTs) have been isolated from a 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) resistant strain (ZANDS) of Anopheles gambiae. The nucleotide sequences of these cDNA species share 80.2% identity and their derived amino acid sequences are 82.

Hydrogenosomes of physiologically resemble mitochondria.

The anaerobic free-living ciliated protozoon is a grazer in anoxic marine sediments. It does not possess mitochondria, but it does have specialized organelles termed hydrogenosomes which release hydrogen gas. The cationic lipophilic cyanine dye DiOC(3) is an indicator of transmembrane electrochemical potential.

The effects of growth dynamics upon pH gradient formation within and around subsurface colonies of Salmonella typhimurium.

pH measurements made in and around submerged colonies of Salmonella typhimurium grown within a model gelatin gel system using pH-sensitive micro- and macroelectrodes indicated some pH heterogeneity occurring in and around the bacterial colony. Inoculation density, initial pH and glucose concentration were all found to influence colony diameter and metabolism of Salmonella colonies. Colony growth in the presence of glucose, at pH 7.

Escape mutants of HIV-1 proteinase: enzymic efficiency and susceptibility to inhibition.

Genes encoding a number of mutants of HIV-1 proteinase were sub-cloned and expressed in E. coli. The proteinases containing mutations of single residues (e.

Proportional Change and Patterns of Homoplasy: Sanderson and Donoghue Revisited.

The effects of two parameters, (1) the number of taxa and (2) the percentage of characters that change per node (referred to as the amount of proportional change or APC) on the retention index (RI) and the consistency index (CI) were examined. Random data sets were generated with a variety of different combinations of the number of taxa and the APC; five replicates for each unique combination were generated producing a total of 660 data sets. Results suggest that both the number of taxa and the APC are significantly correlated with the CI.

The Thatcher illusion as a test of configural disruption.

Bartlett and Searcy's recent account for the Thatcher illusion suggests that inversion impairs holistic facial information. This illusion is used to compare the effects of inverting and negating faces. Subjects made a speeded response to whether just the mouth and the eyes of a face have been inverted.

The hypogastric and thirteenth thoracic ganglia of the rat: effects of age on the neurons and their extracellular environment.

Morphometric analyses of the neurons and microvessels of perfusion-fixed hypogastric (HG) and 13th thoracic (T13) ganglia have been performed in male Wistar rats aged 4, 24 and 30 mo. Estimations of HG volume employing the Cavalieri principle have also been performed and showed that the size of the aged HG is increased by 42%. Routine histological staining of the ganglia with Masson's trichrome indicated that this may be due to the increased amount of interstitial connective tissue which was apparent in the aged animals.

A scanning electron microscope investigation of the effects of pollutants on the hepatopancreatic ceca of Gammarus pulex (L.).

Scanning electron microscope studies were performed on the peripheral muscle network (myoepithelium) of the hepatopancreatic ceca of Gammarus pulex (L.) exposed to lindane, copper, or 3, 4-dichloroaniline (3,4-DCA) for a 24-hr period. The structural integrity of the myoepithelium of gammarids from control and test treatments was assessed using semiquantitative techniques.

Flicker sensitivity as a function of spectral density of external white temporal noise.

Foveal flicker sensitivity at 0.5-30 Hz was measured as a function of the spectral density of external, white, purely temporal noise for a sharp-edged 2.5 deg circular spot (mean luminance 3.

Tendon cells in vivo form a three dimensional network of cell processes linked by gap junctions.

Tendons respond to mechanical load by modifying their extracellular matrix. The cells therefore sense mechanical load and coordinate an appropriate response to it. We show that tendon cells have the potential to communicate with one another via cell processes and gap junctions and thus could use direct cell/cell communication to detect and/or coordinate their load responses.

The characteristics of arginine transport by rat cerebellar and cortical synaptosomes.

The uptake of L-[3H]arginine into synaptosomes prepared from rat cerebellum and cortex occurred by a high-affinity carrier-mediated process. The uptake of arginine appeared to be potentiated by removal of extracellular Na+, inhibited by high levels of extracellular K+, but not by depolarization with veratridine or 4-amino pyridine. The effect of Na+ removal or K+ elevation did not seem to be due to changes in intracellular Ca2+ or pH.