Search PubMed⌕ Search

Biomedical subjects

W S Hu

Publications and source records attributed to W S Hu.

At least 91 records · Page 5Linked to original sources
PubMed1990

Flow cytometric study of hybridoma cell culture: correlation between cell surface fluorescence and IgG production rate.

Flow cytometry was applied to measure the fluorescence of IgG molecules on hybridoma cell surface stained with FITC-conjugated anti-mouse IgG F(ab')2. Using light scattering to exclude the dead cell population from the analysis, the surface fluorescence intensity allows for the differentiation between producing and nonproducing cells. A linear correlation has been found between the intensity of mean surface fluorescence of the cell population and the specific antibody production rate.

Animals↗
PubMed1990

Genetic consequences of packaging two RNA genomes in one retroviral particle: pseudodiploidy and high rate of genetic recombination.

Retroviruses contain two complete viral genomic RNAs in each virion. A system to study in a single round of replication the products of virions with two different genomic RNAs was established. A spleen necrosis virus-based splicing vector containing both the neomycin-resistance gene (neo) and the hygromycin B phosphotransferase gene (hygro) was used. Two frameshift mutants were derived from this vector such that the neo and the hygro genes were inactivated in separate vectors. Thus, each vector confers resistance to only one selection. The vectors with frameshift mutations were separately propagated and were pooled to infect DSDh helper cells. Doubly resistant cell clones were isolated, and viruses produced from these clones were used to infect D17 cells. This protocol allowed virions containing two different genomic RNAs (heterozygotes) to complete one round of retroviral replication. The molecular nature of progeny that conferred resistance to single or double selection and their ratio were determined. Our data demonstrate that each infectious heterozygous virion produces only one provirus. The rate of retroviral recombination is approximately 2% per kilobase per replication cycle. Recombinant proviruses are progeny of heterozygous virions.

Animals↗
PubMed1989

The effect of intravenous captopril on the left ventricular function-an acute study by hemodynamics and radionuclide angiography.

Acute hemodynamic effects of intravenous captopril were measured, and left ventricular function was analyzed by radionuclide angiography in 11 patients with moderate to severe hypertension. All subjects received 1 mg captopril bolus injection initially, and followed in 10 minutes intervals by 2 mg or 5 mg injections according to their blood pressure response. Eight patients experienced an acute reduction of blood pressure; three patients remained unresponsive. The onset of action occurred within 5 minutes with peak effect at 5 to 10 minutes and a duration of action of 20 to 30 minutes. The hemodynamic effects were characterized by arteriolar dilation (reduced systemic vascular resistance P less than 0.01), and apparent venous dilatation (reduced right atrial, pulmonary wedge pressure P less than 0.05 & 0.01 respectively). Pulmonary vascular resistance was not altered. Ejection fraction remained unchanged. Isovolumic relaxation period lengthened (109 +/- 20.1 vs 137.39 +/- 21.15 msec, P less than 0.05) and coronary perfusion pressure gradients dropped (113.8 +/- 5.6 vs 101.1 +/- 11.3 mmHg P less than 0.001) following intravenously administered captopril. Intravenous captopril may reduce arterial pressure by inducing arterial dilatation with minimal adverse effect. The prolongation of isovolumic relaxation period should mainly attribute to the decrease of coronary filling load.

Captopril↗
PubMed1988

Recombinational resolution in primate cells of two homologous human DNA segments with a gradient of sequence divergence.

Human alpha-thalassemia-2 genotype -alpha 4.2 is the result of meiotic recombination between two 1.3 kb long, homologous DNA segments, X(alpha 2) and X(alpha 1), located in the adult alpha globin locus. The two segments can also undergo intramolecular recombination on extrachromosomal vectors transfected into mitotically dividing primate cells (COS 7). The existence of a gradient of sequence divergence between X(alpha 2) and X(alpha 1) makes them an interesting system to study the relationship between efficiencies of homologous DNA recombination and the extent of dispersed and localized base mismatches. By partial restriction mapping and DNA sequencing of plasmids recombined in COS 7 cells and rescued from bacteria HB 101, we have determined the distribution of recombinational resolution sites along the two X blocks. Most, if not all, of the homologous recombination events between the two X blocks appear to be single crossing-over without efficient gene correction or repair of base mismatches. The distribution of the sites of recombinational resolution is inversely correlated with that of the gradient of sequence divergence, with only approximately 7% of the X recombinants resolved within the 3' third of the X blocks where two diverged Alu family repeats reside. The Alu sequence within which one of the X recombinants resolved is homologous to a previously characterized alpha thalassemia deletion point.

Animals↗
PubMed1987

Reconstruction of human alpha thalassemia-2 genotypes in monkey cells.

The human adult alpha globin genes, alpha 2 and alpha 1, are contained within two tandemly arranged duplication units. Each unit spans 4 kb of DNA, and contains three homology blocks (X, Y, Z) separated by non-homologous sequences. Segmental DNA recombination processes between the two units have resulted in high frequencies of two types of deletions in certain human populations, each deletion removing one alpha globin gene from chromosome 16, (alpha-thalassemia 2). In order to study the molecular mechanisms of alpha-thalassemia 2, and of homologous DNA recombination in general in mammalian cells, we have reconstructed these two alpha-thalassemia 2 genotypes in monkey cells. The two duplication units have been cloned in an SV40 origin-containing vector, and transfected into COS 7 cells. Newly replicated plasmid DNA was isolated and analyzed by Southern blot hybridization. Homologous DNA recombination occurs with high frequencies (10-20% per kb of homology), and this generates both types of alpha-thalassemia 2 deletions on the episomes in the monkey cells. Removal of the 5' end of either one, or both, of the X blocks prior to DNA transfection affects the relative frequencies of the two alpha-thalassemia 2 genotypes in a novel way. We consider and discuss these results in terms of several alternative models. Our data suggest the existence of hot spot(s) for initiation of homologous DNA recombination, or recombination promoting element(s), in a specific region of the human adult alpha globin locus. A DNA sequence that defines the boundaries of the two duplication units, and has been implicated in the initiation of gene conversion of the two X blocks, is contained within this region.

Animals↗
PubMed1987

Left ventricular diastolic function in hypertrophic cardiomyopathy: assessment by radionuclide angiography.

Diastolic left ventricular function was studied in 20 patients with obstructive hypertrophic cardiomyopathy, 10 with apical hypertrophic cardiomyopathy, and 20 control subjects. The left ventricular time-activity curve was filtered using Fourier expansion with 4 harmonics. The first and second derivatives of the filtered curve were used to define various parameters of the left ventricle. Isovolumic relaxation period and time to peak filling rate were prolonged in obstructive hypertrophic cardiomyopathy and apical hypertrophic cardiomyopathy (101.56 +/- 44.81 msec, 105.19 +/- 45.27 versus 72.19 +/- 29.36 (CS) P less than 0.05; 169.70 +/- 50.73 msec, 173.83 +/- 50.53 versus 138.02 +/- 29.54 (CS) P less than 0.05, respectively). Rapid filling period, diastasis, atrial contraction period, time to peak filling rate-isovolumic relaxation period, and filling fraction of diastasis were not significantly different in these groups. Fasting filling fraction was 11% lower in hypertrophic groups than in control subjects. The filling fraction of atrial contribution was 12% higher in hypertrophic groups. We conclude: Isovolumic relaxation period, rapid filling phase, and diastasis atrial contraction period can be clearly defined by this technique. Impaired relaxation is an important determinant of decreased left ventricular filling in patients with obstructive hypertrophic cardiomyopathy and apical hypertrophic cardiomyopathy. Prolonged time to peak filling rate is mainly due to increased isovolumic relaxation period in hypertrophic patients. Atrial contraction contributes to more stroke volume in patients with hypertrophic cardiomyopathy.

Adult↗
PubMed1987

Effect of glucose on the cultivation of mammalian cells.

Most of the glucose consumed by mammalian cells cultivated in vitro is converted to lactate. The glucose consumption rate appears to be affected by glucose concentration. In a batch cultivation of cells the glucose concentration can be manipulated at a low level by programmed feeding of glucose. In such a culture the specific consumption rate of glucose and the fraction of glucose converted to lactate can be reduced. This reduced conversion rate of glucose to lactate appears to coincide with an increased oxygen uptake rate. A possible consequence of such programmed feeding of glucose is the increased oxidation of glutamine and the concurrent increased production of ammonium. For the cultivation of hybridoma cells high concentrations of ammonium and lactate can be growth inhibitory. It is suggested that the identification of the optimum cultivation conditions is necessary if such a programmed feeding is to be used to increase cell concentration and medium utilization efficiency.

Ammonia↗
PubMed1986

DNA supercoiling of recombinant plasmids in mammalian cells.

We have used chloroquine/agarose gel electrophoresis and a blot-hybridization technique to study the modulation of superhelicity of extrachromosomal DNA in mammalian cells. The high sensitivity of the procedure has allowed us to measure the change in the specific linking difference or superhelical density (sigma) of a plasmid, psvo alpha 1p3d, after its introduction into COS-7 cells by DNA transfection. Because the molecular weight of psvo alpha 1p3d is approximately the same as that of simian virus 40 (SV40) DNA, the latter can be used as a standard for estimating the average linking difference or number of superhelical turns (tau) of psvo alpha 1p3d after separation of the different supercoiled species on chloroquine/agarose gels. It was found that transfection of monkey cells with either fully supercoiled psvo alpha 1p3d isolated from bacteria (tau = -27 +/- 1, sigma congruent to -0.051) or its relaxed form after treatment with DNA topoisomerase I yields psvo alpha 1p3d samples of the same tau and sigma values of -20 +/- 1 and -0.038, respectively. The difference between the tau values of psvo alpha 1p3d and SV40 in COS-7 cells, in which both plasmids undergo rounds of replication, corresponds to an average difference of 5 +/- 1 superhelical turns. Plasmid psvo alpha 1p3d remains at this lower level of superhelicity for at least 72 hr. The distribution in linking numbers of the topoisomers of psvo alpha 1p3d isolated from transfected COS cells is also more heterogeneous than that of SV40 DNA. These results suggest that the regulation of DNA supercoiling and chromatin assembly may be closely associated with specific DNA sequences. The approach presented here should have a wide application in the study of the regulation and functional role(s) of DNA supercoiling of plasmids in mammalian cells.

Animals↗
PubMed1985

Analysis of tissue-specific methylation patterns of mouse mammary tumor virus DNA by two-dimensional Southern blotting.

We used a two-dimensional Southern blotting procedure to analyze the tissue-specific methylation patterns of the five endogenous mouse mammary tumor viruses in the GR/A mouse strain. Our findings suggest that in certain tissues (brain, kidney, and liver) all proviruses are extensively methylated. In other tissues (spleen, placenta, and testes) all proviruses are hypomethylated to some degree. In these tissues individual proviruses display both quantitative and qualitative differences in methylation. We interpret the general patterns of tissue-specific hypomethylation in terms of a "hitch-hiker" model: mouse mammary tumor virus proviral methylation patterns reflect the tissue-specific activity of neighboring sequences. The observation that certain sites on particular proviruses are differentially methylated in a tissue-specific fashion may reflect tissue-specific differences in the makeup or conformation, or both, of proviral-containing chromatin.

Animals↗
PubMed1984

Mouse mammary tumor virus: specific methylation patterns of proviral DNA in normal mouse tissues.

The methylation state of endogenous mouse mammary tumor virus (MuMTV) proviral DNA was examined in normal mouse tissues. DNAs from various tissues were cleaved with the methylation-sensitive enzymes HhaI and HpaII and analyzed by Southern blotting. Tissue-specific MuMTV proviral DNA methylation patterns were found in the BALB/c, C3H, C57BL, GR/A, and GR-Mtv-2- mouse strains. MuMTV proviral DNA was hypomethylated in DNAs from the spleens and testes of all strains examined. The GR/A mouse strain, which was most thoroughly studied, also showed hypomethylation of MuMTV proviral DNA in bone marrow and placental tissues. Analysis of RNAs extracted from GR/A liver, mammary tumor, testes, placenta, and spleen tissues demonstrated that MuMTV proviral hypomethylation need not reflect significant proviral transcription.

Cell Transformation, Viral↗
PubMed1980

Lidocaine: effect on phagocytosis and purification of monocytes in bovine peripheral blood.

Enumeration of bovine monocytes was determined by latex ingestion, using live cell and fixed cell techniques. Approximately 7.33% of the mononuclear leukocytes were found to phagocytize latex particles by live-cell technique and approximately 7.38% of mononuclear leukocytes phagocytized latex particles by fixed-cell technique. Lidocaine (30 mM) significantly inhibited phagocytosis of latex particles; however, the effects of lidocaine can be reversible. Adherent monocytes can be removed with lidocaine treatment and can be enriched to as much as 90% pure monocytes. Removal of lidocaine was associated with a return to normal cell function.

Animals↗