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Biomedical subjects

H J Woo

Publications and source records attributed to H J Woo.

At least 19 recordsLinked to original sources

Freezing and orientational order in weakly anisotropic fluids.

A simple theoretical method of studying the effect of weak anisotropy on the freezing of classical fluids is discussed. Free energy of the solid phase is separated into contributions due to the formation of the regular lattice and the remaining orientational part. The former is calculated by the density-functional theory, while a mean-field theory of orientational order is developed for the latter. An application to the freezing of hard dumbbell fluids yields results in good agreement with simulations.

Journal Article↗

Simultaneous organic and nitrogen removal from municipal landfill leachate using an anaerobic-aerobic system.

An anaerobic-aerobic system including simultaneous methanogenesis and denitrification was introduced to treat organic and nitrogen compounds in immature leachate from a landfill site. Denitrification and methanogenesis were successfully carried out in the anaerobic reactor while the organic removal and nitrification of NH4+,-N were carried out in the aerobic reactor when rich organic substrate was supplied with appropriate hydraulic retention time. The maximum organic removal rate was 15.2 kg COD/m3 d in the anaerobic reactor while the maximum NH4+-N removal rate and maximum nitrification rate were 0.84kg NH4+-N/m3/d and 0.50kg NO3--N/m3/d, respectively, in the aerobic reactor. The pH range for proper nitrification was 6-8.8 in the aerobic reactor. The organic compounds inhibited nitrification so that the organic removal in the anaerobic reactor could enhance the nitrification rate in the following aerobic reactor. The gas production rate was 0.33 m3/kg COD and the biogas compositions of CH4, CO2, and N2 were kept relatively constant, 66-75, 22-32, and 2-3%, respectively.

Aerobiosis↗

Evaluation of pH inhibition effect on activated sludge by the pseudo toxic concentration (CPT) concept model.

It is generally accepted that the inhibition effect of pH on activated sludge follows the non-competitive inhibition kinetics. But the non-competitive inhibition kinetic equation cannot be directly applied to pH inhibition, due to the difficulty in quantification of pH as a term of inhibitor concentration. So, many empirical equations were developed especially for acidic condition to describe pH inhibition effect. In this research, the pseudo toxic concentration (CPT) concept model to quantify pH inhibition effect on activated sludge was proposed and compared with other existed models. Prediction of performance, presented by Prediction Accuracy and Prediction Accuracy Index, showed that the CPT concept model can explain the reduction of the maximum specific growth rate (mu max) more accurately than any other models do at a wide range of pH. The CPT concept model was applicable not only to activated sludge but also to specific microorganism groups, such as nitrifier, less acidophilic species and nitrifying biofilm.

Bacteria↗

Anticancer activity of hydrophobic peptides from soy proteins.

An anticancer peptide from soy protein was purified and isolated. Defatted soy protein was hydrolyzed with thermoase and hydrophobic peptides were extracted with ethanol. The peptide extract was fractionated by XAD-2 hydrophobic, gel filtration chromatography, and different C18 HPLCs. Anticancer activity of each fraction was assayed by measuring in vitro cytotoxicity on P388D1, a mouse monocyte macrophage cell line. IC50 value of a peptide fraction from Sephadex G-25 chromatography was 0.16 mg/ml. This peptide fraction at 1 mg/ml significantly affected cell cycle progression by arresting P388D1 at G2/M phases. Finally purified peptide from analytical C18 HPLC was nonapeptide of which molecular weight was 1157 Da and the sequence was X-Met-Leu-Pro-Ser-Tye-Ser-Pro-Tyr.

Amino Acid Sequence↗

Inhibition of interleukin-10 (IL-10) production from MOPC 315 tumor cells by IL-10 antisense oligodeoxynucleotides enhances cell-mediated immune responses.

Interleukin-10 (IL-10) has both inhibitory and stimulatory effects on diverse cell types of the immune system. It inhibits the antigen-presenting capacity of monocytes/macrophages and stimulates T cell proliferation. Although many tumors spontaneously release IL-10, the physiological relevance of this phenomenon to the in vivo antitumor immune response is not known. To elucidate the physiological role of tumor-released IL-10, we used IL-10-specific antisense oligodeoxynucleotides (AS-ODN) for the inhibition of IL-10 production from the tumor cells. Incubation of MOPC 315 plasmacytoma with IL-10 AS-ODN in vitro resulted in inhibition of IL-10 production and also in enhancement of the expression of major histocompatibility complex (MHC) class I, MHC class II, and B7-1 molecules. MOPC 315 cells incubated with IL-10 AS-ODN (MOPC-IL10AS) for 16 h in vitro showed reduced tumorigenicity in Balb/c mice. The mice implanted with MOPC-IL10AS effectively rejected the tumor graft, and showed strong cytotoxic T lymphocyte (CTL) activity against the parental MOPC 315 cells. In addition, MOPC-IL10AS were more effective as stimulator cells in mixed lymphocyte/ tumor cell culture, and as target cells in a CTL assay. These results imply that IL-10 spontaneously released from MOPC 315 cells inhibits their immunogenicity and that the inhibition of IL-10 production by IL-10 AS-ODN may be a way to enhance the host cellular antitumor immune response.

Animals↗

Generalized atrophic benign epidermolysis bullosa--poor prognosis associated with chronic renal failure.

Generalized atrophic benign epidermolysis bullosa (GABEB) is an autosomal recessive form of junctional epidermolysis bullosa, with milder clinical features than the Herlitz subtype. A 25-year-old man presented with the clinical and histological findings of GABEB. At the initial visit, laboratory tests revealed that he also had chronic renal failure (CRF). Usually, GABEB has a good prognosis. However, in this case, the patient had CRF as an associated complication. He died of an intracranial haemorrhage combined with sepsis after 3 weeks of hospitalization. This case suggests that renal complications can occur in this relatively mild form of epidermolysis bullosa, and may contribute to morbidity and premature mortality.

Adult↗

No significant correlation exists between core promoter mutations, viral replication, and liver damage in chronic hepatitis B infection.

Hepatitis B virus (HBV) core promoter mutants have been proposed to contribute to severe liver damage by increasing viral loads, but this has not yet been clearly shown. To examine the effects of core promoter mutants on viral load and liver damage, we first developed a polymerase chain reaction (PCR)-based semiquantitative HBV DNA detection method with a high sensitivity (able to detect as low as 10(3) molecules/mL). Then we cloned 12 predominant core promoter mutants from 41 chronic hepatitis B patients. The in vitro promoter and replication activities of these mutants were similar to those of wild-type virus. However, viral load was highly variable, and this was dependent on individual patients rather than mutant type. In addition, there was no mutant type that showed any unique correlation with alanine transaminase (ALT) levels. Viral load was not significantly correlated with ALT level in both cross-sectional and longitudinal studies. Quantitation of HBV levels also revealed no clear correlation between hepatitis B e antigen (HBeAg) status and viral load. Taken together, these results indicated that the replication activity of core promoter mutants has little effect on viral load, and that viral load does not correlate with the severity of liver damage or with HBeAg status.

Adult↗

Pylephlebitis associated with appendicitis.

Pylephlebitis usually occurs secondary to infection in the region drained by the portal venous system. A most common antesecent focus of infection is diverticulitis and the most common blood isolate is E. coli (54%), followed by Proteus mirabilis (23%). Overall mortality is 32% and most of the patients who had died had severe sepsis prior to the initiation of antibiotic therapy. We describe a case of pylephlebitis which had appendicitis and consequent septic thrombosis of the portal vein and its branches, with dissemination of infection to the liver. The patient had recovered due to timely antibiotic treatment alone and resulted in complete resolution. Early diagnosis and treatment are basic to a favorable clinical course.

Adult↗

Heterogeneous HBV mutants coexist in Korean hepatitis B patients.

Although many hepatitis B virus (HBV) mutants have been found in all open reading frames since the precore defective mutant was initially reported, systematic investigations of diverse HBV mutant populations in hepatitis B patients have not been performed. Therefore, we examined whether heterogeneous mutant populations simultaneously exist in Korean hepatitis B patients. In order to detect hepatitis B virus mutants, we amplified a conserved core region and a surface antigen region of HBV DNA by PCR from sera of 27 Korean chronic hepatitis B patients, and then performed single strand conformational polymorphism analysis followed by DNA sequencing analysis. The results showed that heterogeneous HBV mutants in both regions were present in a single as well as in various hepatitis B patients. Sequence analysis revealed a defective interfering particle with missense mutation in the core region. We also found that two subtypes of adr and adw coexisted in a single patient. In addition, a point mutation causing a stop codon in the surface antigen region was observed. We are further analyzing the clinical implications of HBV mutants to identify their roles in the pathogenesis of chronic hepatic disorders induced by HBV.

Adult↗

307-bp fragment in HOXA7 upstream sequence is sufficient for anterior boundary formation.

The HOX genes are expressed in a positionally and temporally restricted manner involving anteroposterior axial pattern formation during early embryogenesis. Previously, we studied the sequence and function of an upstream regulatory region of the human HOXA7 gene. To identify a critical cis-acting element, a deletion analysis was performed along the human control region (HCR) (about 1.1 kb), which was sufficient for setting the anterior boundary of expression in transgenic mice. We demonstrated that a 307-bp control region contains a cis-acting element(s) specifying an anterior boundary as well as a dorsal-ventral restriction in the neural tube at day 12.5 postconception (p.c.). The distinct anterior limit of expression was noted at the level of C7/T1 in the neural tube and spinal ganglia. In addition, our deletion experiments revealed that the HCR consisted of several cis-acting elements which were individually capable of driving regionally restricted expression patterns in the neural tube and limb buds.

Animals↗

Primers determine the sensitivity of PCR-mediated hepatitis B virus DNA detection and pretreatment of PCR mixture with 8-methoxypsoralen eliminates false-positive results.

Most methods for the diagnosis of hepatitis B virus (HBV) infection largely depend on viral DNA detection by polymerase chain reaction (PCR) or radioimmunological assay of viral antigens or antibodies. The quality assurance program recently established in Europe reported that PCR-mediated HBV DNA detection methods used in many laboratories produced a high rate of false-positive and false-negative results. Thus, we attempted to improve the conditions of current PCR methods for detection of HBV DNA. In the present study, we applied a recently developed method of releasing HBV DNA from virion by NaOH treatment of patient serum. Using four different primer sets specific to the HBV core region, we found that the sensitivity of first-round PCR can be improved by more than two orders of magnitude depending on the primers. The second round of PCR using nested primers was sensitive enough to detect up to 10(-6) pg of the HBV DNA, which is equivalent to approximately 3 copies of the HBV genome. Among the approximately 800 HBV-infected patient sera investigated in our laboratory, more than 60% of the tested samples gave positive results in the first-round PCR. The rate of positive results obtained using our experimental conditions is very high in comparison with other reports. The reamplification of the first-round PCR reaction mixture with the nested primers produced practically 100% positive results. For diagnosis of HBV infection, we routinely used 1 microliter of patient serum, which was found to be optimum in our laboratory. Surprisingly, from 20% of our positive results, even serum diluted to 1/100 (0.01 microliter) produced a stronger signal than 1 microliter. This observation suggests that direct PCR amplification of HBV DNA released from serum by NaOH treatment has to be compensated by other DNA detection methods for correct quantitation. In order to eliminate the false positive signal resulting from the carry-over due to massive screening of a large number of samples, PCR reaction mixture containing 8-methoxypsoralen was exposed to ultraviolet light prior to thermal cycle amplification. This exercise did not decrease the sensitivity of the detection method, but almost completely removed the false positive results caused by contaminated templates. We are in the process of improving PCR-mediated HBV DNA detection methods to attain more reliable and easily applicable methods.

Base Sequence↗

Carbohydrate-binding protein 35 (Mac-2), a laminin-binding lectin, forms functional dimers using cysteine 186.

Carbohydrate-binding protein 35 (CBP35), also known as the macrophage surface antigen Mac-2, is a lactosamine-specific lectin whose extracellular properties include the ability to agglutinate cells and to bind avidly to the basement membrane glycoprotein laminin. Although these and other properties would be facilitated by dimerization of this lectin, previous studies have argued against multimeric forms of this protein. We report here that macrophage CBP35, purified by laminin affinity chromatography, exists as several distinct species (Mr 35,000, 67,000, and 80,000) when analyzed under non-reducing conditions. This unexpected finding prompted us to study the biochemistry of multimerization using recombinant CBP35 (rCBP35). rCBP35 expressed in Escherichia coli forms disulfide-linked homodimers (Mr 67,000). The dimeric form of CBP35 binds to laminin with higher affinity than does monomer and by a lactosamine-dependent mechanism. Site-directed mutagenesis indicated that cysteine 186, the single cysteine residue in CBP35, is required for dimerization. These results raise the possibility that homo- and heterodimeric forms of CBP35 contribute to its postulated functions in cell-matrix interactions and growth regulation.

Amino Acid Sequence↗

Carbohydrate-binding protein 35 is the major cell-surface laminin-binding protein in colon carcinoma.

The interaction of tumor cells with laminin is thought to be critical in invasion and metastasis. We found that an endogenous lectin, carbohydrate-binding protein 35 (CBP-35), is the major laminin-binding protein on human colon carcinoma cells and that its surface expression suggests involvement in metastasis. We identified CBP-35 by laminin-affinity chromatography and immunoblotting. Surface expression of CBP-35 on eight human colon carcinoma cell lines was compared by flow cytometry. Poorly differentiated cell lines and DLD-2, a signet-ring carcinoma cell line, expressed more surface CBP-35 than well-differentiated cell lines. Poorly differentiated cell lines and DLD-2 are characterized as aggressive cell lines because they adhere to and invade through reconstituted basement membrane significantly better than well-differentiated cell lines. These data suggest that CBP-35 is involved in tumor cell-basement membrane interactions and that an increase in CBP-35 surface expression may facilitate metastatic potential of colon carcinoma cells.

Antigens, Differentiation↗

The major non-integrin laminin binding protein of macrophages is identical to carbohydrate binding protein 35 (Mac-2).

Current data indicate that cell adhesion to laminin, the major basement membrane glycoprotein, is mediated by specific integrins, a family of adhesion receptors. In addition, most cell types express a complement of high affinity non-integrin laminin binding proteins (LBPs). Despite considerable effort, the function of these LBPs has not been elucidated. We report here that the major non-integrin LBP of murine macrophages exhibits an Mr of 35,000 and is expressed on the cell surface. Protein microsequencing data revealed that this protein is identical to carbohydrate binding protein 35. This murine galactose-specific lectin is the macrophage antigen Mac-2. Thus, these data suggest that the non-integrin LBPs may contribute to laminin adhesion by a mechanism involving protein-carbohydrate interactions.

Amino Acid Sequence↗

A human macrophage hybridoma producing a cytotoxic factor distinct from TNF, LT, and IL-1.

A stable human macrophage hybridoma was established by somatic cell fusion between human peripheral blood monocyte-derived macrophages and an 8-azaguanine resistant clone of a human histiocytic lymphoma cell line U-937 (clone U-937-F9). The hybrid cell line (F9P) exhibited typical macrophage-like morphology and had 30 more chromosomes than U-937-F9 cells. Its macrophage characteristics were confirmed by the manifestation of intracellular nonspecific esterase, the detection of Mo-2 and LEU-M3 antigens on the cell surface, and the demonstration of phagocytic activity. Furthermore, when stimulated with lipopolysaccharide (LPS), this cell line could secrete a considerable amount of a cytotoxic factor (CTF). Distinct from the hybrid cell line, the parental U-937-F9 cells expressed neither Mo-2 nor LEU-M3 antigens on the cell surface, did not show phagocytic activity, and their culture supernatants did not show cytotoxic activity even after LPS stimulation. The activity of CTF in the culture supernatant of the LPS-stimulated hybrid cells could not be neutralized with anti-tumor necrosis factor, anti-interleukin-1, or anti-lymphotoxin antibodies. The CTF had a relative molecular mass of 45-60 x 10(3) daltons as determined by gel filtration on a column of Superose 12, and an isoelectric point of 5.1. The cytotoxic activity was also induced when the hybrid cells were stimulated with the concentrated supernatants of a human T-cell hybridoma containing macrophage activating factor for cytotoxicity or with LP3 tumor cells which were used as target cells.

Antibodies↗

Functional deficiencies of spleen dendritic cells in autoimmune MRL/lpr mice.

We investigated functional aspects of splenic dendritic cells (DC) obtained from MRL/MpJ-lpr/lpr (MRL/l) mice, and their normal counterparts, MRL/MpJ-+/+ (MRL/+) mice. In vitro antigen-presenting activity of DC obtained from MRL/l mice was impaired compared with that of DC from MRL/+ mice. Another major functional aspect of DC, i.e., stimulator cell activity in the allogeneic mixed lymphocyte reaction (MLR), was also found to be impaired in MRL/l mice. MRL/l mice and MRL/+ mice were also examined for co-operation between macrophages (M luminal diameter) and DC. We found peritoneal resident M luminal diameter obtained not only from MRL/+ mice but also from MRL/l mice released a soluble factor which enhanced the activity of DC from the respective mice. Furthermore, DC from both MRL/l and MRL/+ mice responded to the culture supernatant of a mouse macrophage hybridoma clone, S44, which releases a factor that enhances the antigen-presenting activity of DC, and their activity as stimulator cells in the allogeneic MLR was enhanced. However, the degree of enhancement was less in MRL/l mice than in MRL/+ mice.

Animals↗