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Biomedical subjects

S H Hong

Publications and source records attributed to S H Hong.

At least 145 records · Page 8Linked to original sources
PubMed1996

Nucleotide sequence variation in the hypervariable region of the hepatitis C virus in the sera of chronic hepatitis C patients undergoing controlled interferon-alpha therapy.

Ten patients with hepatitis C virus (HCV) infection (experimental group) were treated with interferon-alpha (IF-alpha). Dosage was six million units per day for one week and then three times a week for another six months. Seven HCV-infected patients (control group) did not receive IF-alpha therapy. The hypervariable region (HVR) of HCV in the sera of patients was amplified by reverse transcription-polymerase chain reaction (RT-PCR), and the variation of amino acid sequence in this region was determined. Serum alanine aminotransferase (ALT) activities in five patients treated for six months with IF-alpha fell to the normal range, when HCV was not detected in the sera of three patients. The nucleotide sequence variation in HVR of HCV in the sera of five patients who responded well to the IF-alpha therapy was relatively less than that in another five patients who did not respond to IF-alpha therapy and those in the control patients. These results indicate that the effectiveness of IF-alpha therapy was related to the sequence variation of HVR of HCV. This may have resulted from the selection pressure by humoral antibodies directed to HVR of HCV. It is concluded that the higher rate of sequence variation in HVR of HCV was compatible with a lower degree of effectiveness of IF-alpha therapy.

Adult↗
PubMed1996

3-Deazaadenosine analogues inhibit the production of tumour necrosis factor-alpha in RAW264.7 cells stimulated with lipopolysaccharide.

The effects of 3-deazaadenosine (DZA), 3-deaza(+/-)-aristeromycin (DZAri) and 3-deazaneplanocin (DZNep) on tumour necrosis factor-alpha (TNF-alpha) production were examined in the mouse macrophage cell line, RAW264.7, stimulated with lipopolysaccharide (LPS). The 3-deazaadenosine analogues inhibited the TNF-alpha production and the inhibition was dependent upon the concentration of the analogue. DZA reduced the level of TNF-alpha mRNA suggesting that DZA acts at a transcriptional step. In contrast, DZAri and DZNep had little effect on mRNA levels for TNF-alpha, implying that these compounds inhibit a post-transcriptional or translational biosynthetic step of TNF-alpha synthesis. The observation that homocysteine (Hcy) potentiated the DZA inhibition of TNF-alpha production and of TNF-alpha mRNA levels suggests that the inhibition of TNF-alpha production may be caused by elevated levels of 3-deazaadenosylhomocysteine (DZAHcy). The results show that the 3-deazaadenosine analogues are potent inhibitors of TNF-alpha production in the RAW264.7 cell line stimulated with LPS and suggest that these analogues may be effective agents for the treatment of diseases in which TNF-alpha plays an important pathogenic role.

Adenosine↗
PubMed1996

Topogenic effect of positively charged N-terminal amino acid in ER translocation of yeast alpha-factor precursor.

Expression of fusion proteins between prepro-alpha-factor and somatostatin (SRIF) in yeast, resulted in the correct processing and secretion of the heterologous 14-amino acid SRIF peptide (1). When the chimeric genes were placed under the control of yeast acid phosphatase (PHO5) promoter, significant amount of an unglycosylated form of the fusion precursor molecule accumulated intracellularly, suggesting disruption of an endoplasmic reticulum-mediated function. We report here that the appearance of the precursor is due to an alteration in the three amino terminal residues of the chimera, i.e., Met-Arg-Phe in native prepro-alpha-factor is changed to Met-Phe-Lys in the hybrids. The unglycosylated precursor represents a population of molecules that are disrupted at an early stage of targeting to or translocation across the endoplasmic reticulum membrane. Our data demonstrate that the N-terminus plays an important role in topogenesis. Furthermore, these results show that translocation and glycosylation can be uncoupled from protein synthesis in vivo, and therefore can be posttranslational events in yeast.

Amino Acids↗
PubMed1995

Characterization of SFP2, a putative sulfate permease gene of Saccharomyces cerevisiae.

The SFP2 gene of Saccharomyces cerevisiae has been characterized. The deduced amino acid sequence contained twelve highly hydrophobic domains and showed 50, 47, 44 and 48% homologies to Neurospora crassa sulfate permease II (CYS14), soybean GMAK170 nodulin, human colon mucosa protein (DRA) and a putative open reading frame (ORF) downstream of Escherichia coli prs (phosphoribosyl pyrophosphatate synthetase) gene, respectively, in the aligned regions. Cells lacking SFP2 were viable and displayed no obvious decrease in their growth rate. Southern blot analysis revealed that SFP2 exists as a single copy in haploid genome. Northern blot analysis showed that SFP2 produced a 2.8-kb transcript which was highly expressed under sulfur derepressing condition. SFP2 mRNA was found to turn over with a half-life of approximately 15 min, which may contribute to the regulation of sulfate permease function, and reached its maximal level in about 22 h after depression.

Amino Acid Sequence↗
PubMed1995

Characterization of uvi15+, a stress-inducible gene from Schizosaccharomyces pombe.

The uvi15+ gene of Schizosaccharomyces pombe is a member of a group of stress-inducible genes transcription levels of which increase in response to DNA-damaging agents or heat shock. It encodes a polypeptide of calculated molecular mass 11641 Da, with no significant sequence similarity to other known heat shock proteins. The steady-state level of the uvi15+ gene product of about 12 kDa was increased by heat shock and canavanine, an amino acid analog. This gene also showed a transient increase in expression as cells moved into diauxic shift phase. Although deletion of the uvi15+ gene did not affect the mitotic growth or thermotolerance of cells, the mutant cells rapidly lost viability in stationary phase and under starvation conditions. These cells also showed a defect in sporulation ability. These results suggest that the uvi15+ gene encodes a stress response protein involved in the maintenance of cell viability during entry into stationary phase or under starvation conditions.

Amino Acid Sequence↗
PubMed1995

Isolation and characterization of the promoter region of the rat DNA topoisomerase II alpha gene.

A genomic DNA clone containing the 5'-end of the rat topo II alpha gene was isolated and the intron/exon structure of a 4.0 kb region encompassing the translation initiation site was determined. Multiple transcription initiation sites were found at positions -128, -110, and -100 bp upstream of the ATG codon. A minimal promoter region extending from -192 to the translation initiation codon was identified on transient expression analysis. This region lacks a TATA motif, is moderately GC-rich and contains a high number of CpG dinucleotides, which is characteristic of a housekeeping gene promoter. The fragment extending to position -242 exhibited maximal promoter activity. Putative regulatory elements were delineated within and immediately upstream of the minimal promoter region. On gel retardation and DNase I footprint analyses, two regions, between positions -195 to -159 which interact with protein factor(s) were identified. The minimal promoter region of the rat topo II alpha gene showed high sequence homology with that of human topo II alpha. In a 250 bp region upstream of the translation initiation site, the sequence identity was about 70%. The basic structure of the regulatory region of the rat topo II alpha gene was found to be similar to that of the human counterpart.

Animals↗
PubMed1995

A 33 kDa Pit-1-like protein binds to the distal region of the human thyrotrophin alpha-subunit gene.

Our previous studies demonstrated that at least two DNA regions with upstream limits between positions -223 to -190 and positions -151 to -135 of the human TSH gene are important for transcriptional regulation by TRH in GH3 rat pituitary cells. The proximal region (-151 to -135 bp) including the cAMP-responsive element (CRE) was required for the induction of the TSH gene by TRH, while the distal region (-223 to -190 bp) containing an element similar to the binding site for the pituitary-specific transcription factor, Pit-1, was necessary to amplify the effects of TRH. To determine whether a pituitary-specific nuclear protein, in addition to the CRE-binding protein, is involved in the molecular mechanism of TRH regulation, a gel retardation assay and Southwestern blot analysis were performed on the distal region with GH3 cell nuclear extracts. GH3 extracts generated a distinct DNA-protein complex that was effectively eliminated in the presence of excess unlabelled DNA fragment, and TRH treatment increased the affinity of protein binding remarkably. Excess Pit-1 DNA-binding sequence from the rat prolactin gene inhibited formation of the complex, but mutation of the Pit-1 consensus sequence in the distal region did not eliminate the complex. In addition, Southwestern experiments showed that a 33 kDa nuclear protein present in GH3 cells bound to this region and its binding affinity was increased slightly 2 h after TRH treatment, with the maximal increase (fivefold) at 3 h, which was similar to the results when using gel retardation. Phosphatase treatment of nuclear protein also resulted in a loss of binding affinity. Taken together, these data indicate that the interaction of a pituitary-specific nuclear protein, identical or closely related to Pit-1, with the distal region may be involved in the TRH stimulation of human TSH gene expression.

Animals↗
PubMed1995

Effects of topical nasal decongestants on histology of nasal respiratory mucosa in rabbits.

The aim of this study was to evaluate histologic changes after long-term administration of the topical nasal decongestants phenylephrine and oxymetazoline. Ninety healthy rabbits were divided into 3 groups for topical administration for 1 week, 2 weeks, and 4 weeks. Each group was subdivided into 3 subgroups by topical administration of phenylephrine. oxymetazoline, and physiologic saline as controls. Each study group thus consisted of 10 rabbits. Phenylephrine was administered by 2 puffs in the left nostril 5 times daily and oxymetazoline by 2 puffs twice a day using metered sprayers giving 0.10 ml in each puff with a dosage used in clinical practice. For statistical significance the Kolmogorov-Smirnov test was used. Light and electron microscopic examination were performed after obtaining nasal respiratory mucosa from the nasal septum of the rabbits. Administration of phenylephrine and oxymetazoline for more than 2 weeks caused histologic changes including ciliary loss, epithelial ulceration, inflammatory cell infiltration and subepithelial edema, and the changes were more pronounced with increasing administration duration of the decongestants. Ciliary loss was prominent in the 4-week phenylephrine and oxymetazoline groups. There were significantly severer changes in the morphologic variables in the decongestant groups compared with the control group according to administration duration (p < 0.05). Dilatation or vacuolization of mitochondria and endoplasmic reticula and vesicles ill the cytoplasm were observed in the 2- and 4-week phenylephrine groups as well as the 2- and 4-week oxymetazoline groups. Mild widening of the intercellular space was observed in the 4-week phenylephrine group. Purulent maxillary sinusitis developed in 6 of 10 rabbits treated for 4 weeks with phenylephrine. The results of this study suggest that the administration of decongestants may cause ciliary loss with subsequent inflammatory changes in the nasal respiratory mucosa.

Administration, Topical↗
PubMed1995

Evidences for possible involvement of Rhp51 protein in mitotic events including chromosome segregation.

To understand the role of the Rhp51 protein in Schizosaccharomyces pombe, we examined the phenotypes of the null mutant for the rhp51+ gene. Unlike Saccharomyces cerevisiae rad51 mutants, S. pombe rhp51 mutants (rhp51delta cells) displayed slow growth and heterogeneity in cell size, indicating perturbation of the cell cycle. Furthermore, many aberrant nuclear structures found in 4',6'-diamidino-2-phenylindole (DAPI)-stained rhp51delta cells and the caffeine hypersensitivity of the mutant cells suggested an involvement of the Rhp51 protein in normal chromosome segregation. These data suggested that the Rhp51 proteins were required for normal cell growth as well as a DNA repair pathway. Moreover, rhp51delta mutants showed a considerable sensitivity to ultraviolet (UV) light-irradiation as well as methyl methanesulfonate (MMS) treatment, indicating that the Rhp51 proteins are involved in both the active excision mechanism of UV-induced DNA damage and recombinational repair in S. pombe. Taken together, we suggest that the role(s) of the Rhp51 protein in S. pombe may be different from those of Rad51 in S. cerevisiae.

DNA Damage↗
PubMed1995

A simple and efficient method for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe.

A simple and efficient procedure was described for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe. The present study demonstrated that the quality and the quantity of S. pombe RNA were increased by substituting phenol/chloroform mixture for phenol as a deproteinizing agent in the first vortexing step and using an ice bath instead of a dry ice-ethanol bath in the freezing step. Additionally, this protocol had the advantage of extracting total RNA without any degradation of S. pombe cells. Furthermore, the high amounts and quality of RNA extracted by this modified procedure enabled us to perform some experiments such as Northern blot, S1 mapping, primer extension, and reverse transcriptase reaction-polymerase chain reaction (RT-PCR) without further RNA purification. We suggest that this procedure is very useful to analyse primary structures and steady-state levels of RNA from S. pombe.

Base Sequence↗
PubMed1994

Isolation of UV-inducible transcripts from Schizosaccharomyces pombe.

Four UV-inducible cDNA clones, UVI15, UVI18, UVI22 and UVI31, were isolated from Schizosaccharomyces pombe by subtraction hybridization. All transcripts of these clones were rapidly induced about 5 to 10 fold within 1 hour after UV-irradiation and the nucleotide sequences of these clones did not showed any significant sequence homology to the known genes in the data bases. Transcripts of UVI18 and UVI31 were induced only by UV-irradiation and those of UVI22 were also induced by alkylating agents, suggesting that inductions of these transcripts are specific responses to DNA damages. However, transcript levels of UVI15 were also increased by other cytotoxic agents including heat shock. These results indicate that UVI15 might be a stress responsive gene.

Amino Acid Sequence↗
PubMed1994

Cloning and sequence analysis of rhp51+, a Schizosaccharomyces pombe homolog of the Saccharomyces cerevisiae RAD51 gene.

A homology (rhp51+) of the RAD51 gene in Schizosaccharomyces pombe was cloned by screening a Sz. pombe genomic library using the 3'-end of RAD51 from Saccharomyces cerevisiae as a probe. As in S. cerevisiae, the sequence of rhp51+ showed two MluI cell-cycle boxes and a putative DNA damage-responsive element in its upstream region. The open reading frame codes for a 365-amino-acid (aa) polypeptide with an estimated molecular mass of 40,555 Da. The deduced aa sequence shows 27, 66, 75 and 80% identity with Escherichia coli RecA, S. cerevisiae Rad51 and the Rad51 homologs from chicken and humans, respectively. The aa sequence encoded by rhp51+ contains A- and B-type nucleotide-binding consensus sequences, as found in other RAD51 homologs. Northern blot analysis showed that rhp51+ encodes a 1.7-kb transcript. Methyl methanesulfonate treatment increased the level of this transcript three- to fivefold. Southern hybridization analysis suggests that a single copy of rhp51+ exists in the Sz. pombe genome.

Amino Acid Sequence↗
PubMed1994

Negative effects of wild-type p53 and s-Myc on cellular growth and tumorigenicity of glioma cells. Implication of the tumor suppressor genes for gene therapy.

Human (U251, U87, U343) and rat glioma cell lines (C6, 9L) were examined by the reverse transcriptase-polymerase chain reaction and subsequent nucleotide sequencing analysis to see whether they express wild type (wt)-p53 or mutated form (mut)-p53 messages. Results showed that U87, U343, and C6 cells expressed wt-p53 messages whereas U251 and 9L cells expressed mut-p53 messages. All these cell lines were transfected with wt-p53 cDNA or the s-myc gene linked to the mouse mammary tumor virus (MMTV) promoter. Of several G418-resistant clones obtained from each transfection, a few expressed the s-Myc or wt-p53 proteins. Independent of mutations in the intrinsic p53 gene, the cellular growth in vitro and tumorigenicity in nude mice of these clones were drastically suppressed, the extent of suppression being correlated with the expression level of the transfected gene. Flow-cytometric analysis demonstrated that both p53 and s-Myc arrested the cell cycle at the G1/S boundary. These data suggest that these genes having negative effects on tumor cell proliferation could be used in gene therapy of gliomas, which are caused by alteration of the p53 gene or by some other genetic change.

Animals↗
PubMed1994

Involvement of a cAMP-responsive DNA element in mediating TRH responsiveness of the human thyrotropin alpha-subunit gene.

TRH is known to stimulate the transcription of the TSH gene in pituitary cells. To examine TRH-responsive elements of the human TSH alpha-subunit gene, we have used transient transfection of GH3 rat pituitary tumor cells. Using this system, TRH treatment stimulated expression of a reporter gene containing 846 base pairs from the 5'-flanking region of the human glycoprotein hormone alpha-subunit gene linked to luciferase. Analysis of 5'-deletions of the alpha-subunit sequence revealed that at least two DNA regions with upstream limits between positions -223 to -190 and positions -151 to -135 are important for regulation by TRH. The more proximal region includes a previously defined cAMP-response element (CRE) while the more upstream region contains an element with sequence similarity to the binding site for the pituitary transcription factor, Pit-1. The TRH responsiveness of each individual region was tested by inserting fragments upstream of a thymidine kinase-luciferase reporter gene. The -151 to -100 region had basal enhancer activity and permitted a 3.4-fold response to TRH. The -223 to -168 region did not permit a TRH response, but possessed basal enhancer activity. The combination of both regions resulted in a 5-fold stimulation by TRH. To assess the contributions of different signal transduction pathways, various combinations of treatments were examined. Combined treatment with TRH and forskolin led to an additive activity. Treatment with TRH plus phorbol 12-myristate-13-acetate resulted in the same level of reporter gene activity as with either agent alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

[Hypotensive effect of dihydrolycorine].

Dihydrolycorine-HCL(DL) is a derivative of lycorine, an alkaloid isolated from Lycoris radiata Herb. DL iv 20, 40, and 80 mg.kg-1 lowered the mean arterial pressure in pentobar-bitone-anesthetized normotensive rats, cats, and conscious rats either normotensive or renovascular hypertensive. In pithed rats DL depressed the hypertensive effect of reaction to electric stimulation to the T7-9 segments of spinal cord. DL also depressed the hypertensive effect of phenylephrine. In rabbit aorta ring and rat anococcygeus muscle, DL antagonized the contractile effect of methoxamine, with pA2 values of 5.93 and 6.35, respectively. These results suggest that DL can block alpha 1-adrenoceptors.

Alkaloids↗
PubMed1992

Increased mucin levels in submandibular saliva from mice following repeated isoproterenol treatment.

Mucin decreases with age in the submandibular glands of mice and can be restored by chronic administration of isoproterenol. Mice were given repeated injections of isoproterenol to determine if the increase of mucin in the submandibular gland leads to elevated levels of mucin in the submandibular saliva. Mucin was significantly elevated in both the gland and the saliva after isoproterenol. Gland contents of mucin and the concentration of mucin in their salivas were positively correlated for both the isoproterenol (p = 0.01) and the control group (p less than 0.01).

Animals↗
PubMed1992

The status of the aged in Korea: are the modern more advantaged?

Using individual modernity as a measure of modernization and decision-making power to assess the status of the aged at the micro level, this study examines how modernity, family environment, and demographic characteristics affect the status of the aged in the extended family. Data are analyzed from a sample of 252 Korean elderly living with one of their married children. In contrast to findings at the societal level, this research indicates that individual modernity is associated with higher status of the aged. Individual modernity may become an important resource in negotiating for decision-making power with adult children.

Aged↗