Evaluation of a Short,On-Plate Formic Acid Extraction Method for Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry-Based Identification of Clinically Relevant Yeast Isolates

This report describes a short, on-plate formic acid (FA) extraction method for the identification of clinical yeast isolates using matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS). A total of 41.1% (78/190) and 63.7% (121/190) of yeasts were identified using species log score thresholds of >2.0 and >1.9, respectively. Overall, 97.4% (185/190) of yeasts were identified in combination with conventional FA extraction.     

Simultaneous identification of prednisolone and its ten metabolites in human urine by high performance liquid chromatography-tandem mass spectrometry

The use of prednisolone and prednisone is prohibited by the World Anti‐Doping Agency (WADA) due to their performance‐enhancing effect. The purpose of the present work was to explore the possibility of identification and detection of various metabolites of prednisolone by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) in excretion study samples. Ten metabolites of prednisolone could be identified namely prednisone (11‐oxo metabolite) [M‐1], 6‐β‐OH‐prednisolone [M‐2], 20‐β‐OH‐prednisolone [M‐3], 20‐α‐OH‐prednisolone [M‐4], 20‐α‐OH‐prednisone [M‐5], 20‐β‐OH‐prednisone [M‐6], 2 tetrahydro epimers of 20‐β‐OH‐prednisolone [M‐7], 2 tetrahydro epimers of 20‐α‐OH‐prednisolone [M‐8], 2 tetrahydro epimers of 20‐β‐OH‐prednisone [M‐9], and 2 tetrahydro epimers of 20‐α–OH‐prednisone [M‐10]. Prednisolone was administered in 10‐, 20‐, and 40‐mg dosage to healthy volunteers to study detection of various metabolites. The parent, M‐1, M‐2, and M‐3 could be detected up to 72 h while rest of the metabolites were detectable up to 24 h after drug administration. The detection of newer metabolites of the drug can further be used for confirmatory purposes. Copyright © 2012 John Wiley & Sons, Ltd.     

Features of correction of phosphate-calcium metabolism disorders in terminal renal failure]

Mineral metabolism was studied in 27 hemodialyzed patients with terminal chronic renal failure (CRF). Because of progressing hyperparathyroidism the initial treatment included calcium drugs, active forms of vitamin D (videchol, oxidevit), almagel, bicarbonate hemodialysis and hemosorption. The condition improved in 8 and stabilized in 12 patients. Conservative treatment was ineffective in 7 patients who consequently underwent total (4 cases) or subtotal (3 cases), parathyroidectomy. The response was achieved in 3, stabilization in 3, the disease advanced in 1 patients.     

A Strain Device Imposing Dynamic and Uniform Equi-Biaxial Strain to Cultured Cells

The objective of this study is to design a new apparatus to allow the control of the magnitude and frequency of dynamic stretch applied uniformly to cells cultured on a silicon elastic membrane. The apparatus is designed to produce equi-biaxial dynamic stretches with area changes ranging from 0% to 55% and frequencies ranging from 0 to 2 Hz. Homogeneous finite strain analysis using triangles of markers was performed to compute the symmetric two-dimensional Lagrangian strain tensor on the membrane. Measurements of strain in both static and dynamic conditions showed that the shear component of the strain tensor (E_rc) was near zero, and that there was no significant difference between radial (E_rr) and circumferential (E_cc) components, indicating the attainment of equi-biaxial strain. Bovine aortic endothelial cells were transiently transfected with a chimeric construct in which the luciferase reporter is driven by TPA-responsive elements (TRE). The transfected cells cultured on the membrane were stretched. The luciferase activity increased significantly only when the cells were stretched by 15% or more in area. Cells in different locations of the membrane showed similar induction of luciferase activities, confirming that strain is uniform and equi-biaxial across the membrane. © 1998 Biomedical Engineering Society. PAC98: 8780+s, 8745-k, 8722-q     

Examining the literature on using tap water in wound cleansing

Practitioners working in the community setting and whose caseload predominantly consists of treating chronic wounds often use tap water as a wound cleanser with the rationale that chronic wounds are already colonised with bacteria. However, there is some controversy, as discussed within this review, of the suitability of tap water as a cleansing agent for acute wounds.     

Identification of CD9 extracellular domains important in regulation of CHO cell adhesion to fibronectin and fibronectin pericellular matrix assembly

CD9, a 24-kDa member of the tetraspanin family, influences cellular growth and development, activation, adhesion, and motility. Our investigation focuses on the hypothesis that the CD9 second extracellular loop (EC2) is important in modulating cell adhesive events. Using a Chinese hamster ovary (CHO) cell expression system, we previously reported that CD9 expression inhibited cell adhesion to fibronectin and fibronectin matrix assembly. For the first time, a functional epitope on CD9 EC2 that regulates these processes is described. Binding of mAb7, an EC2-specific anti-CD9 monoclonal antibody, reversed the CD9 inhibitory activity on CHO cell adhesion and fibronectin matrix assembly. This reversal of cell phenotype also was observed in CHO cells expressing CD9 EC2 truncations. Furthermore, our data showed that the EC2 sequence (173)LETFTVKSCPDAIKEVFDNK(192) was largely responsible for the CD9-mediated CHO cell phenotype. Two peptides, (135)K-V(172) (peptide 5b) and (168)P-I(185) (peptide 6a), selectively blocked mAb7 binding to soluble CD9 and to CD9 on intact cells. These active peptides reversed the influence of CD9 expression on CHO cell adhesion to fibronectin. In addition, confocal microscopy revealed that CD9 colocalized with the integrin alpha(5)beta(1) and cytoskeletal F-actin in punctate clusters on the cell surface, particularly at the cell margins. Immunoprecipitation studies confirmed CD9 association with beta(1) integrin. The cellular distribution and colocalization of focal adhesion kinase and alpha-actinin with cytoskeletal actin was also influenced by CD9 expression. Thus, CD9 may exhibit its effect by modulating the composition of adhesive complexes important in facilitating cell adhesion and matrix assembly.