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Biomedical subjects

H H Park

Publications and source records attributed to H H Park.

13 recordsLinked to original sources
PubMed2001

Homodimers of mutant tryptophan synthase alpha-subunits in Escherichia coli.

Tryptophan synthase alpha-subunit from Escherichia coli functionally exists as a heterotetramer of alpha(2)beta(2) with beta-subunit. While wild-type and mutant (F139W, T24M/F139W, and T24L/F139W) alpha-subunits were expressed as a monomer from recombinant plasmids in Escherichia coli, T24A/F139W, T24S/F139W, and T24K/F139W mutant alpha-subunits were abnormally expressed as soluble homodimers in addition to monomers. Monomers of dimer-forming mutant alpha-subunits retain high affinity to beta-subunit, high activity in stimulating catalytic activities of beta-subunit, and nearly intact content of secondary structure, indicating that the global structures of these monomers are identical to that of F139W alpha-subunit. However, fluorescence spectra of Trp139 and ANS binding indicate that significant perturbations occur in the mutant proteins. Interestingly, these defective properties of monomers caused by residue replacement were partially repaired by the dimer formation. As a result, it is suggested that dimers may be formed by domain or loop swapping, and that residue 24 may play important role in maintaining on-pathway of alpha-subunit folding.

Anilino Naphthalenesulfonates↗
PubMed2001

Cloning and sequence analysis of the interleukin-8 gene from flounder (Paralichthys olivaceous).

A cDNA library of mRNA from flounder leukocytes stimulated with bacterial lipopolysaccharide (LPS) and hemagglutinin was constructed to clone cytokine genes of this fish. Initial screening of this library with human cytokine gene probes was not productive and clones with inserts of over 400 nucleotides (nt) were randomly sequenced, and a homologue of the vertebrate interleukin-8 (IL-8) gene was isolated. The flounder IL-8 cDNA encompassed 884 nt, including a coding region of 330 nt. Four cysteines characteristic of CXC chemokines were identified at conserved locations in the putative protein. The deduced amino acid sequence showed 36 and 35% sequence identity with counterpart genes in monkey and human, respectively, and 52% sequence similarity with these genes. However, the putative flounder IL-8 amino acid sequence showed 25% identity and 52% similarity to that of lamprey, the only other piscine IL-8 gene that has been cloned. Flounder IL-8 transcripts were detected in the head-kidney and spleen of LPS-injected flounder and leukocytes stimulated with LPS. It was not detected in the muscle or liver of LPS-injected flounder, tissues taken from non-stimulated flounder and non-stimulated leukocytes.

Amino Acid Sequence↗
PubMed2001

Cloning and expression of thermophilic catechol 1,2-dioxygenase gene (catA) from Streptomyces setonii.

Streptomyces setonii (ATCC 39116) degrades various single aromatic compounds such as phenol or benzoate via an ortho-cleavage pathway using catechol 1,2-dioxygenase (C12O). A PCR using degenerate primers based on the conserved regions of known C12O-encoding genes amplified a 0.45-kbp DNA fragment from S. setonii total DNA. A Southern hybridization analysis and size-selected DNA library screening using the 0.45-kbp PCR product as a probe led to the isolation of a 6.4-kbp S. setonii DNA fragment, from which the C12O-encoding genetic locus was found to be located within a 1.4-kbp DNA fragment. A complete nucleotide sequencing analysis of the 1.4-kbp DNA fragment revealed a 0.84-kbp open reading frame, which showed a strong overall amino acid similarity to the known high-G+C Gram-positive (but significantly less to the Gram-negative) bacterial mesophilic C12Os. The heterologous expression of the cloned 1.4-kbp DNA fragment in Escherichia coli demonstrated that this C12O possessed a thermophilic activity within a broad temperature range (up to 65 degrees C) and showed a higher activity against 3-methylcatechol than catechol or 4-methylcatechol, but no activity against protocatechuate.

Amino Acid Sequence↗
PubMed2001

Oxygen transfer characteristics in a pilot scale surface aeration vessel with Simcar aerator.

The volumetric mass transfer coefficient, KLa, was determined by dynamic method in a surface aerated pilot scale squared vessel up to 0.531 m3 equipped with Simcar type impeller. Through surface aeration, the oxygen transfer characteristics were investigated with the variations of operating variables such as stirring speed, impeller diameter, liquid height and power input per liquid volume (P0/V). It was seen from the results of different oxygen concentration absorption that the dynamic method might lead to errors in KLa when air was used for absorption. To provide reliable KLa values measured by dynamic, the KLa data using pure oxygen were used and confirmed with feeding steady-state method (FSM). As expected, KLa depends on P0/V, impeller size and liquid height. However, for Simcar type impeller, the KLa shows linear dependency on P0/V in contrast to majority of correlations reported in the literature which shows KLa variation of (P0/V)0.65 for disk type impeller. Moreover, it was interesting to find that the bubble behaviors inside the vessel computed by computational fluid dynamics (CFD) could explain qualitatively the KLa changes with operating variables. For the purpose of scale-up procedures, the empirical correlations for predicting KLa were developed within +/- 2% accuracy.

Adsorption↗
PubMed2000

IFN-gamma and IFN-alpha posttranscriptionally down-regulate the IL-4-induced IL-4 receptor gene expression.

As Th1 and Th2 cytokines, IFN-gamma/alpha and IL-4 counterregulate diverse immune functions. In particular, IFN-gamma and IFN-alpha have been reported to markedly suppress the IL-4-induced IgE production and type II IgE receptor (FcepsilonRII/CD23) expression. Because modulation of IL-4R may be an important mechanism in the regulation of IL-4 response, we have investigated the effect of IFN-gamma/alpha on IL-4R expression and signal transduction mechanisms involved in this process. In human mononuclear cells and B cells isolated from tonsil or peripheral blood, IL-4 up-regulates IL-4R(alpha) expression at surface protein and mRNA levels, and the IL-4-induced IL-4R(alpha) is significantly down-regulated by both IFN-gamma and IFN-alpha to a similar extent. The inhibitory effects of IFN-gamma/alpha on the IL-4R mRNA expression require a lag period of about 8 h, and are sensitive to cycloheximide treatment, which suggests that the suppressive effect of IFNs on IL-4R gene expression is a secondary response requiring de novo synthesis of IFN-induced factors. Under such conditions that the inhibitory effects of IFNs are observed, IFNs do not affect the IL-4-induced STAT6 activation and IL-4R transcription, as analyzed by EMSA and nuclear run-on assays, respectively. Subsequently, mRNA stability studies have indicated that the action of IFN-gamma/alpha is primarily mediated by an accelerated decay of IL-4-induced IL-4R mRNA. Thus, it appears that, as already shown in the case of the IL-4-induced FcepsilonRII regulation, posttranscriptional inhibition of IL-4-inducible genes by mRNA destabilization is a common mechanism by which type I and II IFNs antagonize the IL-4 response in human immune cells.

Cells, Cultured↗
PubMed2000

Natural and radiation-induced degeneration of primordial and primary follicles in mouse ovary.

The present study deals with the morphological changes of the degenerating primordial and primary follicles induced by gamma-radiation. Prepubertal female mice of 3 weeks old ICR strain were gamma-irradiated with the dose of LD(80(30)) (8.3 Gy). The ovaries were collected at 3, 6 and 12 h after irradiation. The largest cross-sections were prepared by histological semithin sections for microscopical observations. The ratio (%) of normal to atretic follicles decreased with time after the irradiation in primordial follicles and in primary follicles as well. At 6 h after irradiation, the number of degenerated primordial follicles increased. Germinal vesicles disappeared and lipid droplets increased in number. Granulosa cells became round in shape and apoptotic cells started to appear. The ooplasmic membrane was not recognizable. The ratio of normal to atretic primordial follicles in the control group was 62.5. Then it became lower with time after the irradiation. It went down to 51.6, 49.0, 11.1 and 7.1 at 0, 3, 6 and 12 h, respectively. The ratio of normal to atretic primary follicles in the control mouse ovary was 81.3. It was 80.0, 75.0, 45.5 and 33. 3 at 0, 3, 6 and 12 h after irradiation, respectively. It is concluded that the ionizing radiation acutely induces the degeneration of primordial and primary follicles. The pattern of degeneration is one of the following: (1) apoptosis of one or more granulosa cells with a relatively intact oocyte, (2) apoptosis of an oocyte with intact follicle cells, or (3) apoptotic degenerations of both kinds of cells. These results can provide morphological clues for the identification of the degenerating primordial and primary follicles in normal and irradiated mouse ovaries.

Animals↗
PubMed1999

Effectiveness of preoperative transarterial chemoembolization in presumed inoperable hepatoblastoma.

PURPOSE: To evaluate the effectiveness and therapeutic role of preoperative transarterial chemoembolization (TACE) of hepatoblastoma. MATERIALS AND METHODS: Four patients (one boy, three girls) with unresectable hepatoblastoma were treated twice with preoperative TACE in an effort to improve the surgical and clinical outcome. The patients ranged in age from 8 to 27 months (mean, 15 months). The first TACE was performed superselectively in tumor feeding arteries. The second TACE was performed 3 weeks later. Surgical hepatic resection was performed 1 month after the second TACE. Contrast-enhanced computed tomography (CT) was used to evaluate changes in size, volume, internal texture, and margin of the masses. The toxicity of the chemotherapeutic drugs was evaluated by blood chemistry analysis (AST/ALT, alpha-FP) performed before and after TACE, and after surgery. RESULTS: TACE allowed subsequent surgical resection in all four patients, who remained disease free 16-52 months after operation. There were no major problems related to TACE. There was no chemotherapeutic agent toxicity from TACE. The average largest diameters and volumes of the tumors decreased by 31% (8.3 to 5.6 cm) and 69% (317 to 93 cm2), respectively. CONCLUSION: TACE provided subsequent successful surgical resection and good long-term results in all four patients. The hepatoblastomas were initially considered inoperable because of extensive hepatic involvement and indistinct margins.

Antineoplastic Agents↗
PubMed1998

Induction of quinone reductase by a methanol extract of Scutellaria baicalensis and its flavonoids in murine Hepa 1c1c7 cells.

The effect of extracts of scutellariae radix (Scutellaria baicalensis Georgi) and its flavonoids, baicalin, baicalein and wogonin, on induction of quinone reductase (QR) in the Hepa 1c1c7 murine hepatoma cell line was examined. A significant and dose-dependent induction of QR activity was observed in the methanol extract of scutellariae radix and baicalin. HPCL analysis showed that baicalin was contained as a main component in the methanol extract of scutellariae radix, indicating that baicalin may be the major active principle of QR induction mediated by scutellariae radix extract. To elucidate the mechanism of baicalin-mediated induction of QR enzyme activity, the effect on QR mRNA levels in Hepa 1c1c7 cell cultures was investigated. Using reverse transcriptase-polymerase chain reaction techniques, time- and dose-dependent induction of QR mRNA levels by baicalin were demonstrated in Hepa 1c1c7 cells. On the basis of these results, the scutellariae radix extract or baicalin can be regarded as a readily available, promising, novel cancer chemopreventive agent.

Animals↗
PubMed1995

Modulated expression of type X collagen in Meckel's cartilage with different developmental fates.

Mammalian Meckel's cartilage undergoes regionally diverse histodifferentiation: the caudal end of Meckel's cartilage extends to the developing ear and gives rise to malleus and incus through endochondral ossification while its major distal region differentiates into sphenomandibular ligament and the anterior ligament of the malleus tympanic plate through fibrous transformation. Since the entire Meckel's cartilage develops up to chondrocyte hypertrophy, the regional extracellular matrix components in the hypertrophic Meckel's cartilage may differ in association with the diverse developmental fates. In this project, the expressions of cartilage collagens were investigated in developing rat Meckel's cartilage and particular interest was given to type X collagen. A cDNA, HP114, encoding the NC1 domain of rat alpha 1(X) collagen was cloned, and a synthetic peptide based on the sequence deduced from HP114 was used to generate a monospecific antibody. In situ hybridization of newborn rat condylar and angular cartilages undergoing endochondral ossification showed restricted labeling with the alpha 1(X) collagen probe in the hypertrophic chondrocyte layer. In contrast, the alpha 1(X) collagen probe totally failed to label the major distal portion of Meckel's cartilage even in the hypertrophic cartilage zone. Immunohistochemistry using the anti-type X collagen monospecific antibody consistently failed to recognize the epitope in the corresponding portion of Meckel's cartilage throughout the experimental periods of gestational Day 17, newborn, and Postnatal Day 7, while the strictly localized positive staining was found in the posterior part of Meckel's cartilage which gave rise to malleus and incus. Since major cartilage collagens type II and type IX were found to be present throughout Meckel's cartilage, we postulate that the regulatory molecular mechanism of type X collagen expression may be closely associated with the developmental fates of fibrous transformation and endochondral ossification in mammalian Meckel's cartilage.

Amino Acid Sequence↗
PubMed1991

Transplantation of vascularized composite mandibular allografts in young cynomolgus monkeys.

In the search for donor tissue for massive craniofacial defects, the transplantation of somatic tissue allografts was explored. Four young, out-bred cynomolgus monkeys were the recipients of orthotopic hemimandibular allotransplants from nonrelated cynomolgus monkeys. The transplant consisted of one-half of the mandible with attached muscle, skin, and mucosa. Cyclosporine 15 mg/kg/day was given subcutaneously each day. The 4 monkeys were observed for 13, 27, 63, and 65 days, respectively. All transplants showed primary wound healing and hair growth. The 2 longest survivors chewed, ate a normal diet, and gained weight. Two allografts showed severe rejection signs at 2 to 3 weeks, and the monkeys were euthanized. One monkey had a second episode of rejection that could not be reversed, and it was killed. The fourth monkey died of undetermined causes.

Animals↗
PubMed1991

Growth of vascularized composite mandibular allografts in young rabbits.

Forty-one hemimandible allografts were transplanted in young rabbits immunosuppressed with cyclosporine. The majority of the grafts demonstrated normal wound healing, and growth of hair, bone, and teeth. The mandibular body and the premolars showed significant growth in length. The allografted mandibles functioned sufficiently that the rabbits took oral nourishment soon after surgery. Long-term survival was limited by a toxic "wasting syndrome" specific for rabbits under treatment with cyclosporine.

Animals↗
PubMed1989

Growth of forelimb allografts in young rabbits immunosuppressed with cyclosporine.

Seventeen forelimbs were transplanted orthotopically from young Dutch rabbits to young New Zealand rabbits treated with cyclosporine. The transplanted limbs demonstrated significant bone growth. The growth in the transplanted limbs was about 75 to 80% of that observed in the unoperated limb. The long bones of the 3 longest surviving rabbits (133 days, 150 days, 150 days) studied radiographically demonstrated increases in length over their original lengths (humerus 22%, ulna 26%, and radius 31%). Hair and nail growth were noted at about day 10. Response to pain stimuli (withdrawal of forelimb) and functional use (ambulation with 50% weight bearing) was seen at two to three months. Permanent survival was not achieved because of a species-specific toxic wasting syndrome from cyclosporine.

Animals↗