Glycoprotein B of human herpesvirus 8 is a component of the virion in a cleaved form composed of amino- and carboxyl-terminal fragments

Human herpesvirus 8 (HHV-8) or Kaposi\u27s sarcoma-associated herpesvirus (KSHV) is the only known human member of the Rhadinovirus genus of the gammaherpesvirus subfamily. Antibodies against peptides representing portions of the amino-and carboxyl-termini of HHV-8 gB were produced and used to detect gB expression in Vero cells transfected with the gB gene, in the HHV- 8-harboring cell line, BCBL-1, and in purified virions. Expression of gB was detected in approximately 3% of uninduced BCBL-1 cells, while up to 30% of the cells expressed gB after 12-O-tetradecanoylphorbol-13-acetate (TPA) induction of virus replication. Indirect immunofluorescence assays and confocal microscopy showed that gB was distributed throughout the cytoplasm of BCBL-1 cells and transfected Vero cells. Immunoblot analyses of virion preparations revealed the presence of full-length as well as two smaller than full-length gB-derived species corresponding to the amino- and carboxy- terminal portions of gB, respectively. Biochemical analysis of the gB carbohydrate moieties using glycosylation inhibitors revealed that gB contained N-linked oligosacharides of the high-mannose type, characteristic of precursor carbohydrate chains added in the endoplasmic reticulum. (C) 2000 Academic Press

EVALUATION OF A FOURIER-TRANSFORM-BASED PATTERN-RECOGNITION ALGORITHM FOR TWO-DIMENSIONAL FLUORESCENCE DATA.

A pattern-recognition algorithm for two-dimensional fluorescence data previously reported is critically evaluated. The three spectral matching criteria - sum of the absolute value of the imaginary coefficients of the frequency-domain correlation function, sum of the absolute value of the real-negative coefficients of the frequency-domain correlation function, and the intervector distance between the abbreviated Fourier transforms of two spectra - are calculated. Spectra simulated with a computer as well as data acquired with a video fluorometer are examined. Results indicate that all three parameters are sensitive to changes in peak position, peak width, relative peak height, and intensity of background noises. This work is relevant to identification of bacteria and algae

Two-dimensional fluorescence spectroscopy for bacterial identification

The use of two-dimensional fluorescence spectroscopy in bacterial identification is greatly aided by the development of rapid data acquisition systems and sophisticated pattern recognition algorithms. Numerous approaches using this multiparameter technique have been explored and have proved to be successful in research scale experiments. They are classified as primary or secondary fluorescence methods. Primary methods are limited to those organisms which produce naturally fluorescent pigments while secondary methods are applicable to virtually all bacteria. © 1988

Spectroscopic analysis and drug-binding studies of the CNBr fragments of human serum albumin

Three large fragments (A, B and C) of human serum albumin (HSA) were produced by cyanogen bromide digestion of HSA in order to investigate the specific binding sites. The fragments were isolated by use of gel filtration, followed by high performance ion exchange chromatography. The isolated fragments were examined by use of UV/Vis, steady-state fluorescence, and circular dichroism spectroscopy. The study was extended to examine the interactions of bilirubin and two anionic drugs, warfarin and naproxen, with HSA and the three fragments. The primary bilirubin binding site on HSA molecule appeared to be located between fragment A and fragment C. The results also suggest the binding sites of the two anionic drugs to mosy likely be located in fragment C of HSA molecule. © 1993

Performance Characteristics of the Immunoglobulin G-Capture BED-Enzyme Immunoassay, an Assay To Detect Recent Human Immunodeficiency Virus Type 1 Seroconversion

Recently, we developed an immunoglobulin G (IgG)-capture BED-enzyme immunoassay (BED-CEIA) to identify recent human immunodeficiency virus (HIV) type 1 (HIV-1) seroconversion for use in incidence estimates. We have established an algorithm for its use; developed quality control reagents to monitor the assay; and evaluated its performance for interrun, intrarun, and operator variability. Analysis of 144 individual plates, which involved multiple plate lots and several operators over more than a year, indicated that the coefficients of variation (CVs) were between 10 and 15% for raw optical density (OD) values in the dynamic range between 0.5 and 2.0 OD units; the CVs decreased to 5 to 10% when the OD was normalized (OD-n; OD-n = specimen OD/calibrator OD). The intrarun CVs were generally in the range of 5 to 10% for specimens with ODs <0.5 and less than 5% for specimens with ODs >0.5. The level of concordance between multiple plate lots (n = 6) and multiple operators (n = 7) was quite high (R(2) > 0.9). Comparison of the results of the initial and the confirmatory tests with specimens with OD-n values ≤1.5 demonstrated a high degree of correlation (R(2) = 0.92); 566 (92%) of 615 of specimens tested in the two modes retained the same classification (recent or long-term infection). The values for those specimens with changed classifications (n = 49) were close to the cutoff (OD-n = 1.0), as expected. The twofold difference in the HIV IgG contents between the controls and the calibrator reagents was exploited to monitor individual plate runs by using a control plot, which was incorporated into the spreadsheet for data entry and run monitoring. This information provides baseline data for the successful transfer of BED-CEIA to other laboratories and the use of BED-CEIA for the detection of recent HIV seroconversion and the calculation of incidence estimates worldwide

Selective separation of polar compounds using an electric field coupled normal phase hplc column

An electric field coupled method for separation of polar compounds using normal phase high performance liquid chromatography is described. The method, which is not based on electrophoresis or electrochromatography, selectively changes the column capacity for polar analytes. The use of different columns showed improved column efficiency and up to a 62% increase in the number of theoretical plates. The theory for the improvement is presented. The significance and utility of the method for selective analysis of polar fractions in environmental samples is discussed. Copyright © 1988 by Marcel Dekker, Inc

Selective Separation of Polar Compounds Using an Electric Field Coupled Normal Phase Hplc Column

An electric field coupled method for separation of polar compounds using normal phase high performance liquid chromatography is described. The method, which is not based on electrophoresis or electrochromatography, selectively changes the column capacity for polar analytes. The use of different columns showed improved column efficiency and up to a 62% increase in the number of theoretical plates. The theory for the improvement is presented. The significance and utility of the method for selective analysis of polar fractions in environmental samples is discussed