Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii

Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba

Ductographic Findings of Breast Cancer

Ductography has become the gold standard for the evaluation of patients exhibiting pathologic nipple discharges. In nine patients (age range, 29-67 years; median age, 51 years) with invasive (n=5) or intraductal (n=4) cancer, ductographic findings were recorded, then correlated with mammographic and sonographic findings. Common ductographic findings included complete ductal obstruction, multiple irregular filling defects in the nondilated peripheral ducts, ductal wall irregularities, periductal contrast extravasation, and ductal displacement. Faint microcalcifications or ill-defined masses, which were not opacified by contrast material, were often discovered adjacent to ductal abnormalities. Mammographically and sonographically occult diffusely spreading intraductal cancers often manifested as pathologic nipple discharge. In such cases, meticulous ductographic examinations and interpretations were crucial in order not to miss breast cancers

Interferometric detection of prostate specific antigen based on enzyme immunoassay

AbstractInterferometric detection of Prostate-specific antigen (PSA) based on enzyme immunoassay are investigated. Refractive index changes of substrate are measured for PSA detection. Michelson scheme of optical interferometer was used so as to be applicable to a disposable fluidic chip. When interferometer is used for the measurements of refractive index changes, the detection is over 8 times more sensitive than that of absorbance changes for the same amount of target protein

Atg3-Mediated Lipidation of Atg8 Is Involved in Encystation of Acanthamoeba

Autophagy is a catabolic process involved in the degradation of a cell's own components for cell growth, development, homeostasis, and the recycling of cellular products. Autophagosome is an essential component in the protozoan parasite during differentiation and encystation. The present study identified and characterized autophagy-related protein (Atg) 3, a member of Atg8 conjugation system, in Acanthamoeba castellanii (AcAtg3). AcAtg3 encoding a 304 amino acid protein showed high similarity with the catalytic cysteine site of other E2 like enzymes of ubiquitin system. Predicted 3D structure of AcAtg3 revealed a hammer-like shape, which is the characteristic structure of E2-like enzymes. The expression level of AcAtg3 did not increase during encystation. However, the formation of mature cysts was significantly reduced in Atg3-siRNA transfected cells in which the production of Atg8-phosphatidylethanolamine conjugate was inhibited. Fluorescent microscopic analysis revealed that dispersed AcAtg3-EGFP fusion protein gathered around autophagosomal membranes during encystation. These results provide important information for understanding autophagic machinery through the lipidation reaction mediated by Atg3 in Acanthamoeba