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Biomedical subjects

R C Johnson

Publications and source records attributed to R C Johnson.

At least 487 records · Page 27Linked to original sources

Purine analogue sensitivity and lipase activity of leptospires.

The genus Leptospira can be divided into three groups based on purine analogue sensitivity and lipase (trioleinase) activity. Group 1 contains members of the "parasitic complex" of leptospires which initially cannot grow in media containing 10 mug of 2,6-diaminopurine (DAP) per ml or 200 mug of 8-azaguanine per ml. In addition, leptospires in this group possess lipase activity. Group 2 also contains members of the "parasitic complex" of leptospires. Although these leptospires are similarly sensitive to 8-azaguanine, they differ from group 1 leptospires in that they grow in media containing 10 mug of DAP per ml, and they do not possess detectable lipase activity. Group 3 consists of leptospires belonging to the "biflexa complex." These leptospires are resistant to both purine analogues and have lipase activity.

Adenine↗

Electron microscopy of immune disruption of leptospires: action of complement and lysozyme.

Sequential disruption of the sheath of avirulent leptospires of the serotype canicola with antibody and complement was monitored by electron microscopy. Loosening and separation of the sheath from the protoplasmic cylinder was observed as early as 2 min after exposure to complement. Virulent leptospires of this serotype were morphologically intact after 1 hr of exposure to antibody and complement. Similarly, treatment of leptospires of the serotype patoc with normal serum and complement severely damaged the sheath structure. Removal of the sheath of both serotypes permitted lysozyme to act on the wall of the protoplasmic cylinder. Thus, morphological evidence for the location of the mucopeptide-containing structure of these leptospires was obtained. Viable leptospires with intact sheaths were resistant to lysozyme alone. Sections and negatively stained preparations of sheaths of serotypes canicola and patoc revealed three dense layers with two intermediate light zones and an overall thickness of about 110 A. A periodicity of 40 A was observed in sheath fragments produced by complement. The 70 A wallmembrane complex of leptospires of both serotypes consisted of two dense layers with an intermediate light zone. Structures apparent after removal of the outer sheath included membranous bodies or mesosomes, axial filaments attached to terminal knobs at opposite ends of the cell, and electron-dense intracellular bodies.

Animals↗

Antileptospiral activity of serum. II. Leptospiral virulence factor.

A definite relationship exists between the resistance of leptospires to the antibody-complement system and virulence. Leptospires capable of producing either lethal or renal infections in hamsters or guinea pigs were resistant to the leptospiricidal action of antibody and complement. Avirulent leptospires, in contrast to the virulent organisms, were rapidly immobilized and killed by these serum substances. The change of a virulent culture to the avirulent state as a result of growth in culture media was accompanied by the loss of resistance to antibody and complement. Virulent leptospires were phenotypically altered when grown in the presence of the purine analogue, 8-azaguanine. The cells became sensitive to antibody and complement without a corresponding decrease in virulence. The basis for a leptospiral virulence factor, the ability to multiply in vivo, appears to reside in their capacity to resist the leptospiricidal activity of the host antibody-complement system. The immune leptospiricidal assay provides a simple and rapid method of determining the virulence of a culture.

Animals↗

Differentiation of pathogenic and saprophytic letospires. I. Growth at low temperatures.

The minimal growth temperature of the pathogenic leptospires is between 13 and 15 C. The saprophytic leptospires have a minimal growth temperature between 5 and 10 C, or approximately 5 C below that of the pathogens. The capability of the saprophytic leptospires to grow at temperatures below those which allow the growth of the pathogenic leptospires provides a simple method of discrimination. With an inoculum yielding approximately 8 x 10(7) cells per ml in the test medium and an incubation temperature of 13 C, the saprophytic leptospires were easily differentiated from the pathogenic leptospires. All 13 saprophytic leptospires tested grew in the 10% rabbit serum medium at 13 C, whereas none of the 20 pathogens grew during the 30-day incubation period.

Leptospira↗

Antileptospiral activity of serum. I. Normal and immune serum.

Johnson, Russell C. (University of Minnesota, Minneapolis), and Louis H. Muschel. Antileptospiral activity of serum. I. Normal and immune serum. J. Bacteriol. 91:1403-1409. 1966.-Normal serum was found to exert a leptospiricidal effect, mediated by the complement system, against the nonpathogenic leptospires. Although resistant to normal serum, the pathogenic serotypes were susceptible to antiserum plus complement. Several variables in these immune leptospiricidal reactions were investigated. A reaction period of 3 hr at 37 C between serum substances and 1-day-old cells provided a maximal leptospiricidal effect. The normal serum of the rabbit, guinea pig, bovine, and human were leptospiricidal against the nonpathogenic serotypes, and, in conjunction with rabbit antiserum, rabbit and bovine complement were leptospiricidal against the pathogenic serotypes. Studies with C(14)-labeled leptospires indicated that the immune leptospiricidal reaction was associated with a loss of permeability control. Thus, like the gram-negative bacteria, the treponemes, erythrocytes, and nucleated mammalian cells, the leptospires may be included as cell types susceptible to the antibody-complement system.

Animals↗

Membrane introduction mass spectrometry: trends and applications.

Recent advances in membrane introduction mass spectrometry (MIMS) are reviewed. On-line monitoring is treated by focusing on critical variables, including the nature and dimensions of the membrane, and the analyte vapor pressure, diffusivity, and solubility in the membrane barrier. Sample introduction by MIMS is applied in (i) on-line monitoring of chemical and biological reactors, (ii) analysis of volatile organic compounds in environmental matrices, including air, water and soil, and (iii) in more fundamental studies, such as measurements of thermochemical properties, reaction mechanisms, and kinetics. New semipermeable membranes are discussed, including those consisting of thin polymers, low vapor pressure liquids, and zeolites. These membranes have been used to monitor polar compounds, selectively differentiate compounds through affinity-binding, and provide isomer differentiation based on molecular size. Measurements at high spatial resolution, for example, using silicone-capped hypodermic needle inlets, are also covered, as is electrically driven sampling through microporous membranes. Other variations on the basic MIMS experiment include analyte preconcentration through cryotrapping (CT-MIMS) or trapping in the membrane (trap-and-release), as well as differential thermal release methods and reverse phase (i.e., organic solvent) MIMS. Method limitations center on semivolatile compounds and complex mixture analysis, and novel solutions are discussed. Semivolatile compounds have been monitored with thermally assisted desorption, ultrathin membranes and derivatization techniques. Taking advantage of the differences in time of membrane permeation, mixtures of structurally similar compounds have been differentiated by using sample modulation techniques and by temperature-programmed desorption from a membrane interface. Selective ionization techniques that increase instrument sensitivity towards polar compounds are also described, and comparisons are made with other direct sampling (nonchromatographic) methods that are useful in mixture analysis.

Bioreactors↗

A family study of homeland Korean alcohol use.

Survey data were obtained from 199 homeland Korean families consisting of 199 sets of parents, and 300 college-age sons (162) and daughters (138). Data were obtained regarding quantity and frequency of alcohol consumption by users and of judgments of what constituted "normal" and "problem" alcohol use, flushing status, symptoms experienced following alcohol use, reasons for drinking given by users, and reasons for abstaining given by abstainers. Fathers and sons differed little in use status, but daughters were abstainers far less often than their mothers. Female users drank a good deal less than males, fathers and sons differed slightly, while daughters drank considerably more than their mothers. Own use was substantially correlated with judgments of the nature of normal use and far less related judgements of problem use. There was a significant difference in fast versus slow skin flushing. with a higher proportion of females being fast flushers. Fast flushers drank less than slow flushers and also judged lower amounts of consumption as being "normal." Fast flushers experienced more physical symptoms than slow flushers; persons who drank more experienced fewer symptoms. Males more than females, slow flushers more than fast flushers, and heavy more than light users of alcohol are more likely to endorse a variety of reasons for drinking. Reasons for not drinking differ little across family membership groups. Contemporary homeland Koreans drink less than would have been expected on the basis of prior research, even though there is a substantial generational difference among females, with daughters being less frequently abstainers and drinking more, and more often than their mothers.

Adult↗

Prediction of anxiety disorders using the state-trait anxiety inventory for multiethnic adolescents.

The purpose of this study was to determine the validity of the State-Trait Anxiety Inventory (STAI) in predicting DSM-III-R anxiety disorders based on the Diagnostic Interview Schedule for Children (DISC, Version 2.3) and using Asian/Pacific Islander adolescents. An overall prevalence rate of 9.19% for generalized anxiety disorder, overanxious disorder, or social phobia was consistent with past studies. As hypothesized, STAI negatively worded (i.e., Factor 2) items were better predictors than positively stated (i.e., Factor 1) items. The STAI State mean was a better predictor of concurrent DISC anxiety disorders as compared to STAI State Factors I or 2. In contrast, the STAI Trait Factor 2 (negatively worded) composite was the best predictor for nonconcurrent DISC anxiety disorders as compared to STAI Trait Factor 1 or the overall STAI Trait subscale. Satisfactory predictive-validity values were obtained when using the STAI State mean and Trait Factor 2 composite. Implications of these findings are discussed, including using the STAI as a screening measure for ethnically diverse adolescents.

Adolescent↗

A cheA cheW operon in Borrelia burgdorferi, the agent of Lyme disease.

Borrelia burgdorferi sensu stricto homologues of cheA and cheW were cloned and characterized. A combination of strategies such as polymerase chain reaction (PCR) using degenerate primers, random-primed gene walking PCR and construction of a lamda library were used to identify the putative cheA gene. Sequence analysis of the DNA fragments obtained from the CT strain identified a 2,592-bp open reading frame (ORF) encoding an 864-amino-acid protein with significant similarity (53-64.6% identical residues) to the CheA of several genera of eubacteria. In particular, hallmarks of a histidine kinase family were found such as the location of the histidine autophosphorylation domain very close to the NH2 terminus and the nucleotide-binding site. A second ORF located immediately downstream from the putative borrelial cheA gene encoded a 195-amino-acid protein which displayed a high level of similarity to bacterial CheW. Using reverse transcription PCR, we demonstrated that cheA and cheW form an operon with an upstream, unidentified ORF. The cheA and cheW homologues were located at 722-737 kbp, 738-768 kbp and 743-824 kbp on the linear chromosomes of B. burgdorferi sensu stricto, B. garinii and B. afzelii, respectively. Identification of cheA and cheW is the first step toward elucidation of a possible role of chemotaxis in virulence of the Lyme disease borreliae.

Amino Acid Sequence↗

DNA sequences at the ends of transposon Tn5 required for transposition.

Transposons are a class of genetic elements that can move from one site in a cell's genome to another independently of the cell's general recombination system. Little is known about the mechanism of transposition of compound transposons such as Tn5, but it is thought that a transposon-encoded protein (a transposase) must recognize the outer ends of the element and, together with host factors, catalyse the transfer of the internal DNA into a new site in a manner that may involve replication. It has previously been shown that the synthesis of an IS50R-encoded protein (protein 1) is an essential requirement for Tn5 transposition. Here we demonstrate that a structure containing only the outer 186 base pairs (bp) of both inverted repeats is capable of being efficiently complemented to transpose in Escherichia coli, provided IS50R is located close by on the same replicon. In addition, Bal31-generated deletions indicate that 16-18 bp of the outer end of IS50L are required for transposition. This 16-18-bp sequence contains the 8-9-bp small inverted repeat present at each end of IS50 plus a 9-bp sequence which is homologous to an interrelated sequence present in four copies in the chromosomal origin of replication in a variety of Gram-negative bacteria. This sequence organization suggests that the ends of Tn5 may function to provide a recognition site for the Tn5 transposase adjacent to a sequence recognized by the host replication system.

Base Sequence↗