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Biomedical subjects

M Nguyen

Publications and source records attributed to M Nguyen.

At least 181 records · Page 10Linked to original sources
PubMed1992

1-Deoxymannojirimycin inhibits capillary tube formation in vitro. Analysis of N-linked oligosaccharides in bovine capillary endothelial cells.

Capillary endothelial cells can be induced to form capillary-like structures in vitro by plating on fibronectin-coated dishes (Ingber, D. E., and Folkman, J. (1989) J. Cell Biol. 109, 317-330), thereby mimicking angiogenesis. To assess the role of glycoproteins bearing asparagine-linked oligosaccharides in this process, we tested the effect of oligosaccharide processing inhibitors on the formation of capillary tubes. Deoxymannojirimycin, a compound that prevents synthesis of hybrid and complex-type oligosaccharides, inhibited the formation of capillary tubes. In contrast, swainsonine, an inhibitor that blocks synthesis of complex- but not hybrid-type oligosaccharides, did not inhibit tube formation. Lectin affinity chromatography of 2-[3H] mannose-labeled glycopeptides from endothelial cells induced to form tubes did not reveal a striking difference in the spectrum of oligosaccharides compared to uninduced cells. Since endothelial cells formed tubes normally in the presence of swainsonine, we analyzed glycopeptides from swainsonine-treated induced and uninduced cells. Cells induced to form tubes were enriched in monosialylated hybrid-type oligosaccharides sensitive to alpha-fucosidase, beta-galactosidase, and beta-N-acetylhexosaminidase, suggestive of sialyl Lewis-X determinants. We used an enzyme-linked immunoassay to measure sialyl Lewis-X epitopes on capillary endothelial cells and found that both induced and uninduced cells expressed sialyl Lewis-X epitopes. Deoxymannojirimycin and, to a lesser extent, swainsonine reduced the level of sialyl Lewis-X epitopes in cells induced to form capillary tubes, but neither compound affected the level of epitopes in cell monolayers. We conclude that synthesis of at least hybrid-type oligosaccharides is required for capillary tube formation in vitro and that an increase in monosialylated, fucosylated glycans on asparagine-linked oligosaccharides occurs during this process.

1-Deoxynojirimycin↗
PubMed1992

Transfection of mouse cytotoxic T lymphocyte with an antisense granzyme A vector reduces lytic activity.

Murine CTL have seven serine proteases, known as granzymes, in their lytic granules. Despite considerable effort, convincing evidence that these enzymes play an obligatory role in the lytic process has not been presented. To investigate the function of one of these proteases, granzyme A (GA), we utilized an antisense expression vector to lower the level of the enzyme in the cells. An expression vector containing antisense cDNA for GA and the gene for hygromycin B resistance was constructed and electroporated into the murine CTL line, AR1. Transfectants were selected based on resistance to hygromycin B, and a number of stable lines were developed. One of the antisense lines had greatly reduced levels of GA mRNA, when compared to the parental cells or to control lines transfected with the vector lacking the antisense DNA. The message levels for two other CTL granule proteins, granzyme B and perforin, were unaffected by the antisense vector. The amount of GA, as measured by enzymatic activity, was 3- to 10-fold lower in the transfectant. Most significantly, this line also consistently showed 50 to 70% lower ability to lyse nucleated target cells and to degrade their DNA. Furthermore, it exhibited 90 to 95% lower lytic activity to anti-CD3-coated SRBC. Conjugate formation with target cells, however, was normal. These data provide strong evidence that GA plays an important role in the cytolytic cycle, and that the quantity of enzyme is a limiting factor in these cytolytic cells.

Animals↗
PubMed1992

Addition of a dipeptide spacer significantly improves secretion of ovine trophoblast interferon in yeast.

Yeast has been analysed for its potential to secrete an ovine member of the type-I interferon (IFN) family, trophoblastin (oTP-1). The processing potential of the yeast KEX2 gene product (KEX2p) was evaluated using gene oTP-1 fused to the pre-pro sequence encoding the pre-pro peptide of the yeast alpha-factor precursor. High-level accumulation of nonprocessed (unmatured) recombinant oTP-1 (re-oTP-1) was observed in the medium. In order to short-circuit the limiting activity of KEX2p and to obtain a fully matured re-oTP-1, secretion was directed using a pre::oTP-1 fusion, relying only on signal peptidase-dependent processing. However, secretion of oTP-1 was impaired. High-level secretion was restored when the gene product contained a peptide spacer between oTP-1 and the signal peptidase cleavage site. The oTP-1 variant was shown to have an extended N terminus. An N-extended form was examined further and shown to have the correct size. Surprisingly, the variant retained its in vitro and in vivo biological activities. This system is likely to represent a general method for high-level secretion of type-I IFNs.

Amino Acid Sequence↗
PubMed1992

Expression of the DAF (CD55) and CD59 antigens during normal hematopoietic cell differentiation.

Expression of decay-accelerating factor (DAF or CD55) and of CD59 during hematopoietic cell development in normal bone marrow and on peripheral blood leukocytes were characterized by three-color immunofluorescence experiments. With this technique cell subsets were identified by forward light scatter, orthogonal light scatter, and two cell-surface antigens. For each cell lineage, specific combinations of two monoclonal antibodies labeled with different fluorochromes were used. DAF or CD59 were then quantitated on the defined cell subsets from the fluorescence signal of the respective antibody conjugated with a third fluorochrome. Early uncommitted hematopoietic progenitor cells (CD34+, CD38-) all expressed both proteins homogeneously. Initial commitment to the erythroid (CD71+, CD45dim), myeloid (CD33+), or B lymphocyte (CD10+) lineages was not associated with changes in DAF or CD59 levels. With erythroid development, i.e., after loss of CD45 and decrease of CD71, expression of both proteins decreased. With myeloid maturation, expression of CD59 remained constant and expression of DAF varied. During neutrophil maturation, DAF decreased initially and then reemerged on maturing neutrophils concurrently with the appearance of CD16 (Fc gamma RIII), whereas during monocyte maturation, DAF increased concurrently with up-regulation of CD14. With B cell development, expression of DAF increased concurrently with down-regulation of CD10 and up-regulation of CD20, whereas expression of CD59 diminished slightly late in B cell maturation. Analysis of peripheral blood elements showed that monocytes, neutrophils, and B lymphocytes expressed both proteins homogeneously, but that in contrast to other cell subsets, which all expressed CD59, only a subset of (CD3+) T lymphocytes and (CD16+) Natural killer cells expressed DAF. The absence of DAF was not related to CD4 or CD8 expression or to the presence of activation markers (CD25+, CD38+), memory cell markers (CD58+, CD45RO+), or virgin T cell markers (CD45RA+), but was correlated with expression of CD11b (CR3) and CD11c (gp150/95). Although CD21+ (CR2) and CD35+ (CR1) cells all expressed DAF, CD11a (LFA-1) levels correlated inversely with those of DAF. Although the presence of CD55 and CD59 on early progenitor cells and throughout hematopoietic cell development is consistent with the requirements for both proteins in protection of host cells from complement-mediated injury, the physiological relevance of the unique patterns of variation for each cell lineage is unclear. Nevertheless, the availability of a detailed DAF and CD59 expression map in normal marrow will facilitate analyses of alterations during hematopoietic development that may occur in hematological disorders including paroxysmal nocturnal hemoglobinuria (PNH).

Adult↗
PubMed1992

A comparison of patient risk for pressure ulcer development with nursing use of preventive interventions.

OBJECTIVE: (1) Determine if the Braden scale or Norton scale predicted the same patients to be at risk for pressure ulcer development as were receiving preventive nursing interventions. (2) Identify the items on the Braden and Norton risk assessment scales that the nurses used intuitively to determine a patient's need for a preventive intervention. DESIGN: Cross-sectional study. SETTING: Six hundred-bed, state-supported, long-term care facility. PATIENTS: War veterans who were 82% male and 97% caucasian, mean age 73. MEASUREMENTS: (1) Patients were categorized as at-risk or not-at-risk by the Norton and Braden scales. (2) The presence of a preventive nursing intervention was noted. Agreement in assignment of at-risk status among the two assessments and presence of a preventive intervention was analyzed using Cohen's Kappa. (3) The staff nurses' use of preventive interventions was modeled using stepwise logistic regression. The items from the Braden and Norton risk assessment scales were used as independent variables with staff nurse implementation of a preventive intervention as the dependent variable. RESULTS: Nurse preventive interventions were found on 45% of patients. The Norton scale identified 38% and the Braden scale identified 27% of patients as at-risk. Agreement among the three methods was 0.53. Agreement between the Braden and Norton scales was 0.73. Agreement between use of a preventive intervention and a classification as at-risk by the Braden or Norton scale was 0.41 and 0.43, respectively. Stepwise logistic regression revealed that low Braden mobility scores (Odds Ratio: 2.74) and low Braden friction/shear scores (Odds Ratio: 3.29) were associated with an increased likelihood of a patient receiving a preventive nursing intervention. CONCLUSIONS: The overall level of agreement among the two scales predicting risk and the presence of a preventive intervention was not high. Agreement, however, between the two risk assessment scales was close. The staff nurses apparently relied on a patients' mobility, their exposure to friction/shear, and additional unidentified factors to guide implementation of a preventive intervention. Further study is needed to define the cost, efficacy, and related cost effectiveness of routine pressure ulcer risk assessment.

Aged↗
PubMed1992

Differentiation and maturation of growth factor expanded human hematopoietic progenitors assessed by multidimensional flow cytometry.

Non-adherent cord blood and bone marrow mononuclear cells were analyzed by multiparameter flow cytometry before and at day 2, 4, 7, and 11 of culture in recombinant interleukin 3 (IL-3) and granulocyte colony-stimulating factor (G-CSF, cord blood) or stem cell factor (SCF), IL3 and granulocyte-macrophage colony-stimulating factor (GM-CSF, BM) to assess the differentiation and maturational pathway of myeloid cells. Before cell culture cord blood contained progenitor cells (CD34+) in various differentiation stages (CD38(-)----CD38bright), mature lymphocytes, monocytes, and neutrophils, but no immature neutrophils and immature monocytes. During cell culture, all CD34+ cells acquired the CD38 antigen between day 2 and 5 of cell culture, the CD34 antigen was lost between day 5 and 11 of cell culture. Differentiation of cells into the myeloid cell lineage was characterized by the acquisition of both CD33 and CD71. The latter is indicative for the active proliferation of these cells. Maturation of the cells into the neutrophilic pathway was indicated by the acquisition of first the CD15 antigen followed by CD11b and CD16 respectively. Whereas maturation of the cells into the monocytic pathway was indicated by the acquisition of first CD11b followed by CD14 and a dim expression of both CD15 and CD16. In normal bone marrow, cells of various maturational stages are already present before cell culture. During cell culture differentiation of cells into the myeloid lineage and maturation of the cells along the monocyte and neutrophilic lineage followed identical pathways as was observed before cell culture. Differentiation and maturational pathways of cord blood and adult bone marrow were identical. The results confirm the surface-antigen-defined pathways of myeloid cell differentiation described previously for non-cultured normal bone marrow aspirates. The detailed assessment of cell maturation and differentiation of cultured cells by multidimensional flow cytometry permits the determination of the specific effects of various recombinant human growth factors on myeloid cells.

Antigens, CD↗
PubMed1992

Longitudinal radiologic evaluation of osteoarthritis of the knee.

Radiologic assessment appears to be an objective standard for longterm evaluation of osteoarthritis (OA) and it is inexpensive, quick, simple and noninvasive. We conducted a one year followup study of patients with OA of the knee to evaluate the reproducibility and validity of this evaluation. The intra and interobserver reproducibility of the radiologic variables were evaluated on 275 and 539 patients, respectively, and found to be satisfactory (intraclass coefficient of correlation above 0.70 for the evaluation of the space narrowing of the joint). During the one year followup study, there was a slight but statistically significant deterioration of the joint space narrowing, evaluated on a 6 grade scale on 360 patients (p less than 0.001). The changes in the joint space narrowing were more closely correlated with treatments received by the patients for OA (nonsteroidal antiinflammatory drug (NSAID) intake, synovial fluid aspiration) than with changes in the recorded clinical variables (pain on a visual analog scale, Lequesne algofunctional index). Moreover, some factors such as obesity, generalized OA and flares of OA appeared to be correlated with the deterioration of joint space narrowing. Further studies are necessary to confirm and/or explain the relationship between the deterioration of the joint space narrowing and such factors (i.e., NSAID intake, obesity, flares of OA, generalized OA), which were detected in this study.

Aged↗
PubMed1991

A strongly basic protein of the MAP2 family copolymerizes with tubulin and induces polymerization.

The family of microtubuli-associated proteins of approximately 300 kD molecular weight (MAP2) from porcine brain was fractionated into components of neutral isoelectric point and one polypeptide of strongly basic nature. Both fractions are able to induce the polymerization of purified porcine brain tubulin. In the case of the fractions of an isoelectric point of 7.2, thick and short tubular structures result. Under the influence of the basic protein, extremely long tubules of normal diameter of microtubules are produced. This basic MAP2 copolymerizes with tubulin.

Animals↗
PubMed1991

Effect of BIM-18216, a novel cholecystokinin receptor antagonist, on food intake reduction induced by cholecystokinin.

The role of cholecystokinin (CCK) in food intake was investigated in rats by using BIM-18216, a novel CCK receptor antagonist. In rats fed 6 hr/day, BIM-18216 antagonized the reduction of food intake induced by exogenous CCK octapeptide (CCK-8; 4 micrograms/kg) in a dose-dependent manner and had a maximum effect at 1 mg/kg. BIM-18216 did not antagonize the effect of bombesin on food intake and showed some degree of specificity. BIM-18216 was not able to prevent the effect of endogenous CCK at the beginning of the feeding period. These data demonstrate that BIM-18216 is a potent CCK-specific antagonist. These results also suggest that endogenous and exogenous CCK could act by different regulating pathways.

Animals↗
PubMed1990

Use of a guiding catheter for contralateral femoral artery angioplasty.

We describe a unique method employing a transseptal sheath as a "guiding catheter" that allows contralateral retrograde femoral artery access to perform balloon angioplasty of proximal superficial femoral artery lesions. This technique simplifies arterial access, provides support for crossing lesions, and allows angiographic visualization of target lesions during the procedure.

Aged↗
PubMed1990

1,25(OH)2D3 stimulates phospholipid biosynthesis and surfactant release in fetal rat lung explants.

Lung tissue from 18-day-old rat fetuses was cultured in the presence of 1,25-dihydroxyvitamin D3 - 1,25(OH)2D3 - (10(-9) M) and dexamethasone (10(-7) M) for 48 h. 1,25(OH)2D3 increased the lung content in phospholipids more specifically related to lung surfactant, phosphatidylcholine and phosphatidylglycerol. This increase was similar to that observed with dexamethasone. In addition, unlike dexamethasone, 1,25(OH)2D3 stimulated the surfactant release into luminal spaces, as evidenced by light and electron microscopy. Thus, vitamin D3 might represent an additional factor controlling fetal lung maturation by stimulating phospholipid synthesis and surfactant release from type II cells.

Animals↗
PubMed1989

Anatomical intra-laryngeal anterior branch study of the recurrent (inferior) laryngeal nerve.

The studies are based upon 60 dissections of the recurrent laryngeal n. (inferior laryngeal n.). The authors describe in detail the branches destined for the intrinsic musculature of the larynx. This study is a working approach for interventions of selective laryngeal reinnervation in man. The conclusion is that, it is necessary to resect the inferior cornu of the thyroid cartilage, to reach the abductor and adductor branches of the vocal cord.

Female↗
PubMed1989

Clinical usefulness of estimation of serum fructosamine concentration as screening test for gestational diabetes.

Serum fructosamine levels and fructosamine/protein ratios were measured in 100 pregnant women who underwent glucose tolerance tests because of clinical risk. Compared with normal pregnant women, the 13 study participants with gestational diabetes had higher fructosamine/protein levels (39 +/- 3.9 mumol/gm versus 37 +/- 3.2 mumol/gm, p less than 0.05), fasting serum glucose levels (107 +/- 13.7 mg/dl versus 82 +/- 8.6 mg/dl, p less than 0.001), and area under curve of glucose tolerance test (36 +/- 5 gm x min x dl-1 versus 22 +/- 3.6 gm x min x dl-1, p less than 0.001). The serum fructosamine levels were not significantly different between the two groups of participants (2.3 +/- 0.26 mmol/L versus 2.2 +/- 0.17 mmol/L); 10 of the 13 women with diabetes had a fructosamine/protein ratio within 2 SD of the mean of the groups of normal pregnant women. Spontaneous caloric intakes (r = 0.72, p less than 0.005) and the hospital mean daily capillary glucose levels during diabetic diet (r = 0.72, p less than 0.005) correlated better with the fructosamine/protein ratio than with fasting serum glucose levels (r = 0.58, p less than 0.05) and area under curve (r = 0.57, p less than 0.05). Consequently, serum fructosamine and fructosamine/protein ratio levels should be considered insensitive as a screening test in pregnant patients with clinical risk of gestational diabetes.

Adult↗
PubMed1989

Technical and clinical evaluation of fructosamine determination in serum.

We evaluated a serum fructosamine (glycated serum proteins) assay for efficacy in the diagnosis and follow-up of diabetic patients. A Roche reagent kit, based on nitroblue tetrazolium reduction in alkaline medium, was used in COBAS FARA centrifugal analyzer. We demonstrated that this method is precise, linear and unaffected by serum hemolysis. However, bilirubin affected the test positively and lipemia negatively. Fructosamine (F) correlated positively with total protein (P) (r = 0.809) and albumin (r = 0.746) in a group of 48 non-diabetic individuals. A good correlation was observed between F and glycated hemoglobin from the sera of 514 patients (r = 0.794). A better correlation (r = 0.838) was obtained when F was corrected for P concentration (F/P). Different F and F/P means were calculated only in patients with overt diabetes, compared to normals. Gestational diabetes was associated with a highly significant F increase. However, its low sensitivity (21%) precludes the use of F as an effective screening test for that condition. Nevertheless, because of its simplicity, low cost and rapidity in reflecting changes in the metabolic control of diabetes, F should be considered a valuable test to assess glycemic control in diabetic patients.

Adult↗