Typing of cryptococcal varieties and antifungal susceptibility testing were performed on two strains which were isolated from a nonimmunosuppressed host with cryptococcal meningitis and another from a patient with systemic cryptococcosis with underlying liver cirrhosis. Both varieties of clinical isolates were identified by the use of the glycin-cycloheximide-bromothymol blue agar medium as Cryptococcus neoformans variety neoformans. For the two isolates of Cryptococcus neoformans, the minimal inhibitory concentrations (MIC) of amphotericin B were 0.25 g/mL and the MICs of fluconazole were 8 g/mL.
Agar ; Amphotericin B ; Cryptococcosis ; Cryptococcus neoformans* ; Cryptococcus* ; Fluconazole ; Humans ; Liver Cirrhosis ; Meningitis, Cryptococcal

Background: Delta checks increase patient safety by identifying automated hematology analyzer errors. International standards and guidelines for the complete blood count (CBC) delta check method have not been established. We established an effective, practical CBC delta check method and criteria. Methods: We assessed five delta check methods for nine CBC items (Hb, mean corpuscular volume, platelet count, white blood cell [WBC] count, and five-part WBC differential counts) using 219,804 blood samples from outpatients and inpatients collected over nine months. We adopted the best method and criteria and evaluated them using 42,652 CBC samples collected over two weeks with a new workflow algorithm for identifying test errors and corrections for Hb and platelet count. Results: The median delta check time interval was 1 and 21 days for inpatients and outpatients (range, 1–20 and 1–222 days), respectively. We used delta values at 99.5% as delta check criteria; the criteria varied among the five methods and between outpatients and inpatients. The delta percent change (DPC)/reference range (RR) rate performed best as the delta check for CBC items. Using the new DPC/RR rate method, 1.7% of total test results exceeded the delta check criteria; the retesting and resampling rates were 0.5% and 0.001%, respectively. Conclusions We developed an effective, practical delta check method, including RRs and delta check time intervals, and delta check criteria for nine CBC items. The criteria differ between outpatients and inpatients. Using the new workflow algorithm, we can identify the causes of criterion exceedance and report correct test results.

PURPOSE: To determine whether the expression of mutant myocilin can lead to death of human trabecular meshwork (HTM) cells and to determine whether the mechanism by which this occurs is apoptosis. METHODS: HTM cells were transduced with a recombinant adenovirus expressing human mutant (Q368X) myocilin. The apoptotic death of HTM cells caused by expression of mutant myocilin was examined using a cell proliferation assay, flow cytometry, Western blot analysis, and immunocytochemistry. RESULTS: It appeared that the expression of mutant myocilin itself was not sufficient to cause HTM cell death. Furthermore, the expression of mutant myocilin did not lead to apoptosis of HTM cells although it did elicit a protein unfolding response. CONCLUSIONS: Our data suggest that the mechanism of myocilin glaucoma is not apoptotic death of HTM cells caused by mutant myocilin expression, and that the actual mechanism remains unknown.
Adenoviridae ; Apoptosis ; Blotting, Western ; Cell Death ; Cell Proliferation ; Cytoskeletal Proteins ; Eye Proteins ; Flow Cytometry ; Glaucoma ; Glycoproteins ; Humans ; Protein Unfolding ; Trabecular Meshwork

PURPOSE: To assess whether the expression of heat shock protein 72 (Hsp72) protects rat retinal ganglion cells (RGC-5) from apoptotic cell death. METHODS: Hsp72 expression in RGC-5 cells transduced with replication-deficient recombinant adenovirus was analyzed by Western blot analysis and immunofluorescence. The effect of Hsp72 expression on etoposide-induced apoptotic cell death was examined by microscopic analysis and confirmed by cell proliferation assay. RESULTS: Western blot analysis and immunofluorescence clearly showed adenovirus-mediated Hsp72 expression in RGC-5 cells. Treatment with etoposide resulted in the death of a proportion of the cells by apoptosis. However, this apoptotic cell death was significantly reduced in cells expressing Hsp72, with the reduction in cell death correlating to the level of Hsp72 expression. CONCLUSIONS: Over-expression of Hsp72 alone is sufficient to rescue neuronal cells from apoptotic cell death, suggesting that fine-tuning its expression may be an effective neuroprotective approach in retinal degenerative disease.
Animals ; Blotting, Western ; Cell Death/*genetics ; Cell Survival ; Cells, Cultured ; DNA/*genetics ; Disease Models, Animal ; Etoposide/toxicity ; *Gene Expression Regulation ; HSP72 Heat-Shock Proteins/biosynthesis/*genetics ; Immunohistochemistry ; Rats ; Retinal Degeneration/*genetics/metabolism/pathology ; Retinal Ganglion Cells/drug effects/*metabolism/pathology

Anticoagulants ; Fibrinogen ; Heparin ; Heparin, Low-Molecular-Weight ; Plasma ; Reference Values