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Biomedical subjects

H B Lee

Publications and source records attributed to H B Lee.

At least 19 recordsLinked to original sources
PubMed1997

EGF inhibits expression of WDNM1 and sulfated glycoprotein-2 genes in mammary epithelial cells.

We have previously shown that expressions of ferritin heavy chain (FHC), WDNM1, and sulfated glycoprotein-2 (SGP-2) genes are induced at an involution stage of mammary gland. Here we studied the effect of lactogenic hormones and EGF on the expression of involution-induced genes in HC11 mammary epithelial cells. Insulin, dexamethasone, prolactin, and its combinations did not affect expression of the genes. When cells were cultured in growth medium containing EGF, expression of WDNM1 and SGP-2 genes was strongly inhibited in a dose- and time- dependent manner, whereas expression of FHC gene was not influenced by EGF. Results demonstrate that EGF inhibits expression of WDNM1 and SGP-2 genes in mammary epithelial cells.

Animals↗
PubMed1997

The 5'-upstream region of the rat phospholipase C-beta 3 gene contains two critical Sp1 sites and an HIV Inr-like element.

The 5'-upstream region of the rat phospholipase C-beta 3 gene (PLC-beta 3) has been cloned and characterized. Sequence analysis of the 5'-upstream region showed that it contains a GC-rich region (-166 to +1: 79%) and multiple binding sites for the transcription factors Sp1, AP-1 and AP-2, but does not contain a canonical TATA box. Primer extension analysis of total RNA isolated from rat glial cell C6Bul revealed that single transcription start point (tsp) is located at an initiator (Inr) element similar to that found in the HIV promoter. Gel mobility shift and competitive mobility shift assays indicated that this Inr element forms a DNA-protein complex with the HIV Inr-binding protein, LBP-1/CP2 or a homologue. In order to localize functional elements of the 5'-upstream region of the rat PLC-beta 3 gene, 5'-deletion fragments were cloned into a chloramphenicol acetyltransferase (CAT) reporter vector. Transient transfection analyses of the 5'-deletion mutants identified a crucial promoter element located at -128 to -14. Supershift mobility assays, site-directed mutagenesis and DNase I footprints indicated that Sp1 binds to three GC boxes within the sequence between -128 and -14 of the PLC-beta 3 promoter. Transient transfection analyses of promoter constructs containing site-specific mutation(s) of these three GC boxes demonstrated that two GC boxes, located proximal to the tsp, are important elements for normal promoter activity.

Animals↗
PubMed1997

Plasma protein adsorption and platelet adhesion onto comb-like PEO gradient surfaces.

Comb-like polyethylene oxide (PEO) surfaces were prepared on low-density polyethylene (PE). The comb-like PEO chain density was changed gradually along the sample lengths by corona discharge treatment with gradually increasing power and the following graft copolymerization of poly(ethylene glycol) monomethacrylate macromers (PEO-MA). The macromers with different PEO repeat unit, 1, 5, and 10, were used. The prepared comb-like PEO gradient surfaces were characterized by water contact angle, Fourier transform infrared spectroscopy in the attenuated total reflectance mode, and electron spectroscopy for chemical analysis. All these measurements indicated that the PEO chains are grafted on the PE surface with gradually increasing density of PEO. Plasma protein adsorption and platelet adhesion on the PEO gradient surfaces decreased with increasing PEO chain length and surface density. As observed by scanning electron microscopy, PEO10-MA-grafted surface with high PEO density was very effective in preventing protein adsorption and platelet adhesion and did not activate the platelets.

Adsorption↗
PubMed1997

Development and characterization of an alginate-impregnated polyester vascular graft.

Alignate gels are known to be biocompatible, degradable, and nontoxic. In this study, sodium alginate was impregnated into a porous, knitted polyester graft (Microvel double velour graft) 6 mm in diameter. The alginate-impregnated graft was investigated in vitro and in vivo to evaluate its potential for use as a new vascular graft impervious to blood, while retaining high porosity for tissue ingrowth and biological healing. For in vitro investigation, the coating weight, water permeability, morphology, and mechanical properties of the alginate-impregnated grafts were compared to those of control or commercially available collagen-impregnated (Hemashield) grafts. The water permeability of the controls (1846 mL/min.cm2 at 120 mm Hg) was reduced > 99% by the alginate impregnation, rendering the graft impervious to blood. The coating weight of the alginate was 45 mg/g of graft, producing a much lower value than that of the collagen-impregnated model (310 mg/g). For in vivo investigation, the alginate-impregnated grafts were implanted in the aorta of mongrel dogs without preclotting for scheduled periods ranging from 4 h to 6 months. The control grafts after preclotting and the collagen-impregnated grafts without preclotting were also implanted for 3 and 6 months for comparison. Gross observation of the explanted grafts and histologic examination of the representative sections were conducted for three types of grafts using a light microscope after hematoxylin-eosin staining. No significant differences were observed between the histologic appearance of the alginate-impregnated grafts and that of the preclotted and collagen-impregnated grafts in terms of the degree of inflammation, foreign-body giant cell reaction, and intimal fibrosis. Endothelial-like cells were present on the midsections of all the grafts after 3 months of implantation. The resorption rate of alginate impregnated into the graft was also examined after staining the sections with periodic acid-Schiff reagent, Toluidine blue, and Alcian blue, which are specific for alginates. The staining alginate was partially visible between the graft fabrics up to 1 month after implantation, but was completely resorbed after 3 months. This preliminary study demonstrated that the use of an alginate as a biological sealant instead of proteins such as collagen, gelatin, and albumin may be a feasible approach to developing imprevious textile arterial prostheses, since the proteins have been reported to be generally unstable, hard to obtain in pure forms, not easy to crosslink and control resorption rate, and difficult to render compatible with standard storage and sterilization procedures.

Alginates↗
PubMed1997

Interaction of cells on chargeable functional group gradient surfaces.

Functional group gradient surfaces where the surface density of grafted functional groups changes gradually along the sample length were prepared on low density polyethylene (PE) sheets by corona discharge treatment with gradually increasing power and graft copalymerization of acrylic acid (AA), sodium p-styrene sulphonate (NaSS), and N,N-dimethyl aminopropyl acrylamide (DMAPAA). AA and NaSS are negatively chargeable and DMAPAA is positively chargeable in phosphate buffered saline or cell culture medium at pH 7.3-7.4. The functional group gradient surfaces were characterized by the measurement of water contact angle, Fourier transform infrared spectroscopy in the attenuated total reflectance mode, and electron spectroscopy for chemical analysis. All these measurements indicated that the functional groups were grafted on the PE surfaces with gradual increase of their density. The interaction of Chinese hamster ovary cells with the functional group gradient surfaces along the sample length was investigated. The cells that had adhered and grown on the surfaces were counted and observed by scanning electron microscopy. It was observed that a greater quantity of the cells had adhered and grown onto the positions with moderate density of the functional groups. This may be related to the hydrophilicity of the surface. The DMAPAA-grafted surface showed a large amount of cell attachment probably owing to the positive charge character, while the AA-grafted surface, which is negatively charged, showed poor cell attachment, as expected. The NaSS-grafted surface which is also negatively charged showed a large amount of cell attachment. This may be closely associated with the existence of an aromatic ring close to the ionizable group in NaSS. It seems that surface functional groups and their charge character as well as wettability play important roles for cell adhesion, spreading, and growth.

Amines↗
PubMed1997

Chaperone-like function of lipocortin 1.

Lipocortin 1 (LC1) is a 37 kDa member of the annexin family of proteins. It has been proposed to act as a mediator of some of the actions of glucocorticoids in anti-inflammatory and immune suppressive functions. LC1 has been shown to play a role in cell proliferation, apoptosis, and differentiation. However, the exact biological functions of LC1 still remain obscure. Here it is shown that LC1 displays a chaperone-like function. Stoichiometric amounts of LC1 suppressed thermally induced inactivation and aggregation of the test enzymes citrate synthase and glutamate dehydrogenase. LC1 was also effective in refolding guanine hydrochloride-denatured glutamate dehydrogenase, as judged by circular dichroism spectroscopy.

Animals↗
PubMed1997

Unilateral multilayered musculocutaneous V-Y advancement flap for the treatment of pressure sore.

We have devised a modified technique using the gluteus maximus musculocutaneous flap as multilayered sliding V-Y advancement to cover pressure sores on the sacral area. Nine patients with relatively large (average 7 x 7 cm) sacral grade IV pressure sores underwent unilateral multilayered V-Y advancement flap. All patients were followed for a minimum of 8 weeks. The mean postoperative follow-up was 32.3 months, with a range of 24 to 39 months. Using this technique, the success of surgery, i.e., the percentage of sores that healed, was 100 percent in our patients. The advantages of this technique include sufficient advancement of the flap, coverage of large ulcer defects using only a unilateral musculocutaneous flap, and preservation of the contralateral gluteus maximus muscle for future use.

Aged↗
PubMed1997

Identification of urinary dipeptidase as the released form of renal dipeptidase.

Amphipathic and hydrophilic forms of human renal dipeptidase and urinary dipeptidase were purified by affinity chromatography using cilastatin, a dipeptidase inhibitor, as the ligand. The sequence analyses of the first ten amino acids of renal and urinary dipeptidases were shown to be identical, and they are Asp-Phe-Phe-Arg-Asp-Glu-Ala-Glu-Arg-Ile. Unambiguous results of amino acid sequencing, the molecular weight of native protein (190 kD), the molecular weight of subunit (47.7 kD) and a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicate that the enzymes are composed of homotetramers. This is the most direct evidence that urinary dipeptidase is the released form of renal dipeptidase. In fact, they are the same enzymes.

Amino Acid Sequence↗
PubMed1997

Differences in immunophenotyping of mucosal lymphocytes between ulcerative colitis and Crohn's disease.

OBJECTIVES: Immunologic studies have characterized the numbers and types of inflammatory cells in diseased inflammatory bowel disease (IBD) mucosa but have yielded conflicting results regarding intestinal lymphocytes activation in IBD. We investigated the levels of lymphocytes subsets, interleukin-2 receptor, transferrin receptor, and T cell receptors in mainly isolated lamina propria lymphocytes. Including intraepithelial lymphocytes of normal colonic mucosa or IBD (ulcerative colitis and Crohn's disease) mucosa to understand the pathogenesis of IBD. We have results from this study. RESULTS: 1) In comparing ulcerative colitis with control, IL-2R (p < 0.05), TR (p < 0.01), and CD3/HLA-DR (< 0.05) showed a significant increase. 2) In comparing Crohn's disease with control, CD3 (P < 0.05), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. 3) In comparing Crohn's disease with ulcerative colitis, CD19 (p < 0.01), TR (p < 0.01), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. CONCLUSION: From these results, there are increased T cell markers, IL-2R, TR, and CD3/HLA-DR in UC, but differently, decreased CD3, TCR alpha/beta and TCR gamma/delta in CD compared with control. In addition, definitive differences in lymphocytes markers, CD19, TR, TCR alpha/beta and TCR gamma/delta, which are higher in UC than in CD, may elucidate the different immunopathogenesis between UC and CD.

Adult↗
PubMed1997

Single-sample methods to measure GFR with technetium-99m-DTPA.

UNLABELLED: Many single-sample methods have been suggested to simplify the methodology of glomerular filtration rate (GFR) measurement. The relative accuracy of these competing methods is still not clear for clinical practice. METHODS: Fifty-four GFR studies with 99mTc-DTPA were performed on 37 adult patients (serum creatinine 0.8-10 mg/dl). Each study included a UV/P, plasma clearance method (three-sample) and single-sample methods. The single-sample methods used were those of Christensen and Groth (modified by Watson), Constable, Dakubu, Groth and Aasted, Jacobsson, Morgan, Russell and Tauxe. RESULTS: When the GFR > or = 30 ml/min (n = 26), all of the single-sample methods were highly correlated with UV/P. The correlation of the single-sample method with the plasma clearance was higher than with UV/P. In this group (GFR > or = 30 ml/min), the Groth 4-hr sample method had the best value of both absolute difference and percent absolute difference (mean +/- s.e. = 11.05 +/- 2.51 ml/min and 14.08% +/- 2.43%, respectively). Most single-sample methods do not perform well at GFR < 30 ml/min (n = 28), and none of them has a good correlation with UV/P or plasma clearance at this level of renal function. However, the Groth and Aasted's 4-hr sample method was the best compared with others (mean +/- s.e. = 8.43 +/- 1.30 ml/min for absolute difference, and 65.91% +/- 16.70% for percent absolute difference). CONCLUSION: Single-sample methods may not correctly predict GFR in advanced renal failure. Groth and Aasted's method with 4-hr plasma sample has both the lowest mean absolute difference and percent absolute difference in both the group with GFR > or = 30 ml/min and GFR < 30 ml/min. All methods perform acceptably at GFR > or = 30 ml/min.

Adult↗
PubMed1996

Augmentation of tumor metastasis by platelet-activating factor.

The effect of platelet-activating factor (PAF) on experimental pulmonary metastasis by the B16F10 murine melanoma and the possible involvement of PAF in the activities of tumor necrosis factor alpha (TNF-alpha) and interleukin 1alpha (IL-1alpha) in tumor metastasis were investigated. i.p. injection of PAF enhanced the lung colonization in a dose- and time-dependent manner. PAF enhanced lung colonization when it was administered after, but not before, B16F10 inoculation. Multiple injections of PAF were more effective than a single injection. Neutralization of endogenous PAF with PAF antagonist BN50739 decreased lung colonization, suggesting that endogenous PAF plays an important role in pulmonary metastases. A single i.p. injection of TNF-alpha or IL-1alpha caused a marked enhancement in lung colonization. TNF-alpha- and IL-1alpha-mediated enhancement in lung colonies was significantly inhibited by BN50739. These results demonstrate that PAF has a metastasis-enhancing effect and is a mediator of the metastatic activities of TNF-alpha and IL-1alpha.

Animals↗
PubMed1996

Enhancement of thermostability and catalytic efficiency of AprP, an alkaline protease from Pseudomonas sp., by the introduction of a disulfide bond.

A site-directed mutagenesis in AprP, an alkaline protease isolated from Pseudomonas sp. KFCC 10818 was carried out in order to obtain increased thermostability. Sites for cysteine substitutions to form disulfide bond within AprP were chosen by comparing the sequences with aqualysin I, an alkaline thermostable serine protease whose disulfide bonds seems to be important for its thermostability. Gly199 and Phe236 residues were each replaced with cysteine by site-directed mutagenesis. The G199C/F236C mutant enzyme appeared to form a disulfide bond spontaneously during its expression. It also showed improved kinetic parameters for the hydrolysis of a synthetic peptide substrate at pH 8.5 and 10.5 compared to those of the wild-type enzyme. The half-life of the G199C/F236C mutant was found to be 2 to 4.8 times longer than that of wild-type under various experimental conditions, except when tested under reducing condition, where no significant differences in the half-life of the two types were observed. Therefore, it is concluded that the introduction of the disulfide bond enhanced the thermostability and the catalytic efficiency of the enzyme AprP.

Alkaline Phosphatase↗
PubMed1996

In situ detection of hepatitis C virus RNA in liver tissue using a digoxigenin-labeled probe created during a polymerase chain reaction.

The cellular localization of hepatitis C virus (HCV) RNA in liver tissue was studied by nonisotopic in situ hybridization using a digoxigenin-labeled cDNA probe created during a polymerase chain reaction on samples from 16 patients with chronic HCV infection. Hybridization signals were recognized in the cytoplasm of the hepatocytes, and a few hepatocytes had hybridization signals in the nucleus as well. HCV RNA positive hepatocytes were found in 1 of 9 patients with chronic persistent hepatitis, 2 of 5 patients with chronic active hepatitis, and in each of 2 patients with chronic active hepatitis and cirrhosis. Positive signals were found in many hepatocytes within the lobule in liver sections of patients with advanced chronic active hepatitis. A number of HCV RNA positive hepatocytes were found in nodules, but not in the area of fibrosis. On the other hand, positive signals were found in a few hepatocytes scattered in the lobule in a patient with chronic persistent hepatitis. The mean ALT levels in the patients with positive signal (175.6 +/- 44.2 U/L) were significantly higher than in those without a signal (70.27 +/- 16.1 U/L) (P < 0.05). The findings suggest that a larger amount of HCV may be present during the advanced than during the early stages of type C hepatitis and nonisotopic in situ hybridization using a digoxigenin-labeled HCV cDNA probe created during a polymerase chain reaction deserves wider application for the detection of HCV replication in specimens.

Adult↗
PubMed1996

Blood and cell compatibility of gelatin-carrageenan mixtures cross-linked by glutaraldehyde.

Mixtures of gelatin and iota-carrageenan cross-linked by glutaraldehyde were prepared and their physical properties and blood and cell compatibility were compared to gelatin as a control material. According to scanning electron microscopic observation of fracture surfaces, the mixtures were composed of dispersed and continuous domains which might be generated by phase separation of carrageenan. The thermal degradation temperature of iota-carrageenan in the mixtures rose with increasing gelatin content. The swelling process in the mixtures proceeded slower than in gelatin. Tensile strengths of the mixtures, except that containing 50% iota-carrageenan, increased with increased amounts of iota-carrageenan in the mixtures. The iota-carrageenan contents at the surfaces of the mixtures were generally higher than those admixed originally. Static friction coefficients of the mixtures were lower than that of gelatin. Plasma recalcification times of the mixtures were longer than that of gelatin. Platelet adhesion of the mixtures was lower than that of gelatin, while cell adhesion and growth assays using Chinese hamster ovary cells showed that cell adhesion and growth were not dependent on adding iota-carrageenan. It was concluded that blood compatibility of the mixtures increased and cell compatibility did not decrease, compared to gelatin.

Animals↗
PubMed1996

Prevention of platelet adhesion on the polysulfone porous catheter by saline solution perfusion, I. In vitro investigation.

A new technique for the prevention of platelet adhesion on the polysulfone (PSf) porous catheter system by means of saline perfusion was investigated. Commercially available ultrafiltration PSf hollow fibers were used as porous catheter tubes. Sixty milliliter of citrated canine blood was circulated for 20 min through the annular catheter flow cell system in vitro. Platelet adhesion onto the PSf porous catheter tube decreased sharply with increased saline solution perfusion rate. From the semilogarithmic relationship between the number of adhered platelets and perfusion rate, the critical perfusion rate for preventing adhesion of platelets could be about 0.3 ml min-1 cm-2. After the surface of PSf porous tubes was modified by non-ionic surfactant coating and air plasma treatment for 20 seconds without saline perfusion, the number of adhered platelets was lower than that of the control tube. These modification methods may be more effective in the prevention of initial platelet adhesion if combined with saline perfusion.

Animals↗
PubMed1996

Dialysis in patients with diabetic nephropathy: CAPD versus hemodialysis.

Diabetic nephropathy has emerged as a major cause of ESRD over the past decade, being the most prevalent cause of ESRD requiring dialysis in North America (United States and Canada) and the second highest in the incidence rate in Europe, Japan, Korea, Australia, and New Zealand. A greater proportion of older patients and of patients with diabetic nephropathy and other comorbid conditions has been treated with CAPD. Despite the preferential use of CAPD to treat a high-risk group of patients, the overall and/or selection-adjusted mortality was similar between HD and CAPD groups. Among diabetic patients, selection-adjusted mortality was similar between HD and CAPD or lower in CAPD than in HD, the difference being greatest among younger patients and significant through the age of 52, or higher in CAPD than in HD with higher risk of death for older diabetics (age > or = 50 years), but with similar risk among younger diabetics (age < 50 years). Technique survival was also variably reported as similar between HD and CAPD, lower, or higher with CAPD compared to HD. Diabetic CAPD patients had more hospital admissions and more days in the hospital and higher withdrawal rates from dialysis compared to diabetic HD patients. These disparate results of patient and technique survival between HD and CAPD in diabetic patients may have resulted from patient selection criteria with different comorbid conditions on entrance to dialysis, quantity of dialysis, and other unrecognized factors. Prospective randomized studies are needed to assign a cause-and-effect relationship between the choice of dialysis modality and patient and technique survival among patients with diabetes mellitus as well as with all other diagnostic categories.

Diabetic Nephropathies↗