Serologic differences in strains of Sporothrix schenckii.

To obtain evidence that Sporothrix scheneckii enters the body by contact with contaminated materials, the antigenic property of strains from different sources was investigated. The reciprocal absorption test of the antisera against a soil isolate and a human isolate (KO 4606) showed that the absorbed antisera against KO 4606 possessed unique antigen(s) in addition to the common antigen of both strains. Twenty-three clinical isolates were tested with absorbed antisera. Not all of them possessed the unique antigen(s), but there were serologic varieties among S. schenckii strains, regardless of their sources, clinical type of the disease and the morphology of the yeast phase cells.     

Dark Rearing in the Visual Critical Period Causes Structural Changes in Myelinated Axons in the Adult Mouse Visual Pathway

Neurochemical Research - An appropriate sensory experience during the early developmental period is important for brain maturation. Dark rearing during the visual critical period delays the...     

Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)

A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs) was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. coli. A high antigenicity of recombinant BbTRAP2 (rBbTRAP2) was observed with field B. bovis-infected bovine sera collected from geographically different regions of the world. Moreover, antiserum against rBbTRAP2 specifically reacted with the authentic protein by Western blot analysis and an indirect fluorescent antibody test. Three bands corresponding to 104-, 76-, and 44-kDa proteins were identified in the parasite lysates and two bands of 76- and 44-kDa proteins were detected in the supernatant of cultivated parasites, indicating that BbTRAP2 was proteolytically processed and shed into the culture. Apical and surface localizations of BbTRAP2 were observed in the intracellular and extracellular parasites, respectively, by confocal laser microscopic examination. Moreover, native BbTRAP2 was precipitated by bovine erythrocytes, suggesting its role in the attachment to erythrocytes. Furthermore, the specific antibody to rBbTRAP2 inhibited the growth of B. bovis in a concentration-dependent manner. Consistently, pre-incubation of the free merozoites with the antibody to rBbTRAP2 resulted in an inhibition of the parasite invasion into host erythrocytes. Interestingly, the antibody to rBbTRAP2 was the most inhibitive for the parasite’s growth as compared to those of a set of antisera produced against different recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c), rhoptry-associated protein 1 C-terminal (BbRAP-1CT), and spherical body protein 1 (BbSBP-1). These results suggest that BbTRAP2 might be a potential candidate for development of a subunit vaccine against B. bovis infection.     

A simple and rapid determination of biapenem in plasma by high-performance liquid chromatography

A simple, rapid and precise HPLC method using ultrafiltration to remove plasma protein was developed to determine biapenem concentrations in human plasma. Plasma was separated by centrifugation at 4 degrees C from blood collected in heparinized vacuum tubes, and biapenem was stabilized by immediate mixing the plasma with 1M 3-morpholinopropanesulfonic acid (MOPS) buffer (pH 7.0) (1:1). Biapenem was detected by ultraviolet absorbance at 300nm with no interfering plasma peak. The calibration curve of biapenem in human plasma was linear from 0.04 to 50microg/mL. The limit of detection was 0.01microg/mL, which was more than 40-fold lower than that of conventional plasma protein precipitation using ammonium sulfate. The assay has been clinically applied to pharmacokinetic studies in patients.     

A discriminant model constructed by the support vector machine method for HERG potassium channel inhibitors

HERG attracts attention as a risk factor for arrhythmia, which might trigger torsade de pointes. A highly accurate classifier of chemical compounds for inhibition of the HERG potassium channel is constructed using support vector machine. For two test sets, our discriminant models achieved 90% and 95% accuracy, respectively. The classifier is even applied for the prediction of cardio vascular adverse effects to achieve about 70% accuracy. While modest inhibitors are partly characterized by properties linked to global structure of a molecule including hydrophobicity and diameter, strong inhibitors are exclusively characterized by properties linked to substructures of a molecule.     

A novel type of two-component regulatory system affecting gingipains in Porphyromonas gingivalis

We surveyed the Porphyromonas gingivalis W83 genome database for homologues of FimS, the first two-component sensor histidine kinase, which could possibly control virulence factors. Including fimS, we found six putative sensor kinase genes in the genome. The gene encoding one of the homologues was cloned from a P. gingivalis plasmid library, sequenced, and analyzed using its mutants. Two gene-disruption mutants were created in strain ATCC 33277 by introducing a drug cassette into the gene. The mutants formed nonpigmented colonies, indicating that they might be defective in proteinase production, a characteristic of this organism. Proteinase activities, measured as arginine- and lysine-specific (Rgp and Kgp gingipains, respectively) activities, of the mutants were almost half those of the parent strain. Unlike the parent and wildtype strains, most of the gingipain activities were detected in the culture supernatant, not in cells, of the mutants. Abnormal production of gingipains was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analyses. These results strongly suggest that this newly-discovered two-component sensor kinase is involved in maturation and proper localization of gingipains to the outer membrane through an unknown mechanism. The gene encoding the sensor histidine kinase was designated gppX, which represents regulation (X) of gingipains and black pigmentation in P. gingivalis.