Role of Rilpivirine and Etravirine in Efavirenz and Nevirapine-Based Regimens Failure in a Resource-Limited Country: A Cross- Sectional Study

IntroductionEtravirine(ETR) can be used for patients who have failed NNRTI-based regimen. In Thailand, ETR is approximately 45 times more expensive than rilpivirine(RPV). However, there are no data of RPV use in NNRTI failure. Therefore, we assessed the susceptibility and mutation patterns of first line NNRTI failure and the possibility of using RPV compared to ETV in patients who have failed efavirenz(EFV)- and nevirapine(NVP)-based regimens.MethodsClinical samples with confirmed virological failure from EFV- or NVP-based regimens were retrospectively analyzed. Resistance-associated mutations (RAMs) were interpreted by IAS-USA Drug Resistance Mutations. Susceptibility of ETR and RPV were interpreted by DUET, Monogram scoring system, and Stanford University HIV Drug Resistance Database.Results1,279 and 528 patients failed EFV- and NVP-based regimens, respectively. Y181C was the most common NVP-associated RAM (54.3% vs. 14.7%, p<0.01). K103N was the most common EFV-associated RAM (56.5% vs. 19.1%, P<0.01). The results from all three scoring systems were concordant. 165(11.1%) and 161(10.9%) patients who failed NVP-based regimen were susceptible to ETR and RPV, respectively (p = 0.85). 195 (32.2%) and 191 (31.6%) patients who failed EFV-based regimen, were susceptible to ETR and RPV, respectively (p = 0.79). The susceptibility of ETV and RPV in EFV failure was significantly higher than NVP failure (p<0.01).ConclusionThe mutation patterns for ETR and RPV were similar but 32% and 11% of patients who failed EFV and NVP -based regimen, respectivly were susceptible to RPV. This finding suggests that RPV can be used as the alternative antiretroviral agent in patients who have failed EFV-based regimen.     

Cloning and characterization of the hemA gene for synthesis of δ-aminolevulinic acid in Xanthomonas campestris pv. phaseoli

The gene from Xanthomonas campestris pv. phaseoli that is involved in the C5 pathway of -amino-levulinic acid (ALA) of Escherichia coli. Subcloning of deletion fragments from the initial 2.5-kilobase (kb) chromosomal fragment allowed the isolation of a 1.6-kb fragment that could complement the hemM mutation. Nucleotide sequence analysis of the 1.6-kb DNA fragment revealed an open reading frame that encodes a polypeptide of 426 amino acid residues, and the deduced molecular mass of this polypeptide is 46768 Da. The amino acid sequence shows a high degree of homology of the HemA protein, which is glutamyl-tRNA reductase, to other organisms. Thus, we examined the complementation test of the cloned gene from Xanthomonas with a hemA mutation of E. coli and found that the gene complemented the hemA mutation. These results suggest that the cloned gene is hemA and the gene from Xanthomonas also complements both hemA and hemM mutations, as in the case of the E. coli hemA. Correspondence to: Y. Murooka     

Immunological detection of Salmonella paratyphi A in raw prawns.

A slot blot enzyme-linked immunosorbent assay, using monoclonal antibodies specific only for Salmonella paratyphi A, to detect S. paratyphi A contamination in raw prawns has been established. When artificially contaminated prawn samples were tested. S. paratyphi A contamination could be identified correctly within 20 h. No false positives from samples artificially contaminated by other microorganisms were obtained. The sensitivity was such that as few as 1 S. paratyphi A organism per g of raw prawn could be detected. Therefore, the assay constituted a promising test for the rapid and specific detection of S. paratyphi A in prawns.     

Inactivation of <Emphasis Type="Italic">Burkholderia pseudomallei</Emphasis><Emphasis Type="Italic">bsaQ</Emphasis> results in decreased invasion efficiency and delayed escape of bacteria from endocytic vesicles

Burkholderia pseudomallei, an infectious Gram-negative bacterium, is the causative pathogen of melioidosis. In the present study, a B. pseudomallei strain with mutation in the bsaQ gene, encoding a structural component of the type III secretion system (T3SS), was constructed. This bsaQ mutation caused a marked decrease in secretion of BopE effector and BipD translocator proteins into culture supernatant. The B. pseudomallei bsaQ mutant also exhibited decreased efficiencies of plaque formation, invasion into non-phagocytic cells and multinucleated giant cell (MNGC) development in a J774A.1 macrophage cell line. Co-localization of the bacteria and lysosome-associated membrane glycoprotein-1 (LAMP-1) containing vesicles suggested that defects in MNGC formation may result from the delayed ability of this B. pseudomallei mutant to escape from the vacuoles of macrophages. Veerachat Muangsombut and Supaporn Suparak contributed equally to this work.     

Distribution of Aeromonas hydrophila serogroups in different clinical samples and the development of polyclonal antibodies for rapid identification of the genus Aeromonas by direct agglutination

We characterized a collection of 256 Aeromonas hydrophila strains isolated from blood, discharge and stool for their serogroup designation. Of these, 2.3% were untypable and 15.2% were rough strains. Among the typable strains, about 50% comprised serogroups O:11, O:16, O:18, O:34 and O:83. To develop rapid differentiation of Aeromonas from other oxidase-positive bacteria, antisera against Aeromonas were produced to establish a direct, genus-specific, agglutination test. It was found that among 105 isolates of Aeromonas, 102 showed positive results with the agglutination test. The calculated sensitivity and specificity were 97.1% and 90.7%, respectively.     

Characterization of anaerobic non-spore-forming bacteria in municipal sewage sludge and integrated paper-mill waste water

Summary Forty-three isolates of Gram-negative, mesophilic, non-spore-forming anaerobic cellulolytic bacteria were obtained from (i) an anaerobic reactor treating waste water from an integrated paper mill and (ii) an anaerobic sewage-sludge digestor. These isolates were studied for carbohydrate fermentation and fermentation products. By numerical techniques, 22 isolates could be placed in two groups: group A (10 isolates) and group B (12 isolates). The isolates belonging to group A showed degradation of filter paper in 2–7 days. They were slightly-curved long rods and similar toBacteroides cellulosolvens andAcetivibrio cellulolyticus. Acetic acid was produced as major product. The bacteria also produced ethanol, isobutanol, pyruvic and lactic acids. Group B strains degraded filter paper in 4–5 weeks. They were short rods and produced propionic, lactic, succinic and acetic acids as fermentation products. The remaining 21 isolates could disintegrate filter paper in 2–5 weeks. They showed variable fermentation patterns, both as to fermentable carbohydrates and end products. Except for one isolate, which showed obvious similarity toButyrivibrio fibrisolvens, the isolate differed distinctly from reference strains of ruminai origin.

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Caractérisation des bactéries anaérobies non-sporulantes dans la boue résiduaire de station d'épuration des eaux domestiques (boues d'égout) et dans l'eau résiduaire d'une usine intégrale de pâte à papier

Résumé 43 souches de bactéries cellulolytiques Gram-négatives, mésophiles, non-sporulantes, et anaérobies ont été isolées à partir (A) d'un réacteur anaérobie traitant l'eau résiduaire d'une usine intégrale de pâte à papier et (B) de digesteurs anaérobies de boues d'égout. Ces souches ont été étudiées quant à leurs propriétés de fermenter les hydrates de carbone et quant à leurs produits de fermentation. Des techniques numériques ont permis de classer 22 de ces souches dans deux groupes: le groupe A (10 souches) et le groupe B (12 souches). Les souches appartenant au groupe A dégradaient le papier filtre en 2 à 7 jours. Elles se présentaient sous la forme de longs bâtonnets curvilignes et ressemblaient àBacteroides cellulosolvens et àAcetivibrio cellulolyticus. Le metabolite majeur était l'acide acétique. Elles produisaient aussi de l'éthanol, de l'iso-butanol, et des acides pyruvique et lactique. Les souches appartenant au groupe B dégradaient le papier filtre en 4 à 5 semaines. Elles se présentaient sous la forme de bâtonnets courts et produisaient, comme métabolites, les acides propionique, lactique, succinique et acétique. Les 21 souches restantes dégradaient le papier filtre en 2 à 5 semaines. Elles présentaient des profils de fermentation variables tant en ce qui concernait les hydrates de carbone fermentés que les produits finaux. A part une souche qui présentait une similitude évidente avecButyrivibrio fibrisolvens, ces souches différaient de façon marquée d'avec les souches de référence du rumen.

     

The rpoE operon regulates heat stress response in Burkholderia pseudomallei

Burkholderia pseudomallei is a gram-negative bacterium and the causative agent of melioidosis, one of the important lethal diseases in tropical regions. In this article, we demonstrate the crucial role of the B. pseudomallei rpoE locus in the response to heat stress. The rpoE operon knockout mutant exhibited growth retardation and reduced survival when exposed to a high temperature. Expression analysis using rpoH promoter-lacZ fusion revealed that heat stress induction of rpoH, which encodes heat shock sigma factor (sigma(H)), was abolished in the B. pseudomallei rpoE mutant. Analysis of the rpoH promoter region revealed sequences sharing high homology to the consensus sequence of sigma(E)-dependent promoters. Moreover, the putative heat-induced sigma(H)-regulated heat shock proteins (i.e. GroEL and HtpG) were also absent in the rpoE operon mutant. Altogether, our data suggest that the rpoE operon regulates B. pseudomallei heat stress response through the function of rpoH.