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Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica.

Entamoeba histolytica can act as a harmless commensal organism in the lumen of the large intestine, or can cause invasive amoebiasis. Some workers have suggested that there are two distinct subspecies of this organism, and that only one of these is associated with invasive disease. Present isoenzyme tests to identify the subspecies take several days to analyse: we report a technique that uses immunofluorescence with monoclonal antibodies, takes two days to perform, and may, therefore, assist in the clinical management of patients infected with this organism.

Animals

Molecular clones from a non-acutely pathogenic derivative of SIVsmmPBj14: characterization and comparison to acutely pathogenic clones.

Molecularly cloned simian immunodeficiency viruses capable of inducing acute, fatal disease in pig-tailed macaques had been derived previously from a biological clone (bcl-3) of the PBj14 isolate of SIV from sooty mangabey monkeys (SIVsmmPBj14). The present study was undertaken in order to characterize virus from a second biological clone of SIVsmmPBj14, bcl-1, which fails to induce acute or fatal disease. Polymerase chain reaction was used to amplify 5' and 3' viral genome halves. The DNA sequence of two 3' halves was determined, and an infectious recombinant generated using a bcl-3-derived 5' half and a bcl-1-derived 3' half. Overall, bcl-1- and bcl-3-derived viruses displayed close homology, differing by a total of 2% at the DNA level and 1-6% at the amino acid level within the 8 open reading frames examined. In contrast to the bcl-3-derived viruses, the bcl-1-derived viruses encode a truncated transmembrane envelope glycoprotein. Another consistent difference was the presence of a 22 bp duplication in the U3 portion of the long terminal repeat (LTR) of bcl-3-derived viruses that includes the NF-kappa B transcriptional enhancer binding site. To assess the importance of this duplication, virus chimeras were generated which removed the duplication from the 3'-LTR or from both LTRs of a bcl-3 clone. The former virus was unstable, reacquiring the duplication through recombination with the 5' LTR. No consistent difference were observed, however, between viruses with or without the duplication in the in vitro studies conducted to date.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence

Differences in aerosol survival between pathogenic and non-pathogenic strains of Legionella pneumophila serogroup 1.

A strain of Legionella pneumophila serogroup 1 known to be virulent for guinea-pigs was found to be least stable at a relative humidity (r.h.) of 60% when stored as a small particle aerosol. Three L. pneumophila serogroup 1 strains of different virulence for guinea-pigs were then tested at a r.h. of 60% at 20 degrees C. The most virulent strain was found to have the best survival and the avirulent strain was least stable. The strain of intermediate virulence did not survive as well as the virulent strain but was more stable than the avirulent strain. Strains of L. pneumophila serogroup epidemiologically associated with legionnaires' disease had better survival in small particle aerosols than strains which were not associated with disease. Subtyping with monoclonal antibodies also showed that the type more commonly associated with disease survived longer in aerosols than the other subtypes.

Aerosols

Flotation of mastitis pathogens with cream from subclinically infected quarters. Prospects for developing a cream-rising test for detecting mastitis caused by major mastitis pathogens.

Bacterial isolates, originating from 36 subclinically infected quarter milk samples, were labelled with 75Se and checked for cream-rising at various temperatures in a system analogous to the ABR test ("Abortus Bang Ringprobe"; the cream-rising test based on stained brucella organisms for detection of brucellosis). Diagnostic specificity and sensitivity were analyzed in experiments where labelled bacterial isolates were mixed with a number of quarter milk samples with known bacteriological status as well as samples from healthy control quarters. Creaming at 37 degrees C resulted in specific "recognization" as the bacterial isolates showed preferential flotation in the milk samples from which they had been isolated as well as is milk samples harbouring the same bacterial species. At lower creaming temperatures, the specificity was lost since all the isolates became concentrated in the cream phase irrespective of the milk sample. When comparing the specific recognization by cream of the respective bacteria, bacterial species vary: The prospects for developing diagnostic cream-rising tests for Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli seems promising, but less so for coagulase-negative staphylococci, Streptococcus dysgalactiae, and Streptococcus uberis. The mechanism behind the cream-rising of labelled bacteria at 37 degrees C seems to lie in specific fat globule membrane-bound immunity of IgA type. Therefore the milk fat globules from chronically infected quarters function as absorbents for the respective isolates. Flotation of bacteria with cream indicates an in vivo mechanism enabling bacteria to invade the upper parts of milk ducts within the udder.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Relationship between growth inhibition and mitochondrial function in petite-negative yeasts. I. Effects of antibiotics and dyes upon pathogenic and non-pathogenic Candida species.

Antibiotics and dyes which preclude growth of Saccharomyces cerevisiae in media containing oxidizable carbon sources arrested the growth of Candida albicans, Candida tropicalis and Candida utilis even in glucose medium. The growth in the presence of sub-inhibitory concentrations of the various antibiotics and dyes determined a reduction in the cell survival but with no accumulation of respiratory deficient mutants. Under these culture conditions, the total respiration declined leaving a residual antimycin A-resistant--hydroxamate-sensitive O2 uptake, and the amount of the respiratory cytochromes aa3 and b synthesized was reduced. SDS gel electrophoresis of soluble proteins prepared from the antibiotic-treated cells showed some bands in the MW range 92-100 K, which became faint after the cells were grown in the presence of some mitochondrial inhibitors. The ultrastructural analysis of these cells evidenced disappearance of the mitochondrial cristae and their replacement by unfolded membranes. The data obtained suggest that the petite negative trait of Candida could depend on the non-viability or on the very low viability of those cells which have lost their mitochondrial function.

Anti-Bacterial Agents

Relationship between growth inhibition and mitochondrial function in petite-negative yeasts. II. Effects of central nervous system drugs upon pathogenic and non-pathogenic Candida species.

Six nervous system drugs which inhibited vegetative reproduction of S. cerevisiae arrested also mitotic division of C. utilis, C. albicans and C. tropicalis. Chlorpromazine and chlorpheniramine which proved to be the most effective, affected respiration and cytochrome biosynthesis. Electrophoretic bands with MW congruent to 100 K were faint in silver-stained electrophoregrams of proteins from cells grown in the presence of a sub-inhibitory concentration of chlorpromazine.

Amitriptyline

In vitro nail invasion by pathogenic and non-pathogenic fungi under different culture conditions.

Data from the literature suggest that the nutritional environment can modify major metabolic functions of fungi and possibly their aggressivity towards keratinous structures. Trichophyton rubrum (TR), Microsporum canis (MC), Scopulariopsis brevicaulis (SB), Aspergillus fumigatus (AF) and Penicillium spec. (P) were inoculated on media of different nutritional value, in presence of nail fragments. The activity of the fungi was evaluated at two and four weeks for intensity and depth of invasion of nail samples. Nail invasion was most pronounced by MC, especially when grown on rice agar or peptone agar. Nail invasion by the other fungi tested was less important, Sabourand glucose and rice agar were most favorable. Our results indicate that nutritional factors can, at least in some fungus species, alter their rate of nail invasion.

Aspergillus fumigatus