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H G Kang

Publications and source records attributed to H G Kang.

At least 19 recordsLinked to original sources

Molecular cloning and characterization of a dermatan-specific N-acetylgalactosamine 4-O-sulfotransferase.

We have identified and characterized an N-acetylgalactosamine-4-O-sulfotransferase designated dermatan-4-sulfotransferase-1 (D4ST-1) (GenBank(TM) accession number AF401222) based on its homology to HNK-1 sulfotransferase. The cDNA predicts an open reading frame encoding a type II membrane protein of 376 amino acids with a 43-amino acid cytoplasmic domain and a 316-amino acid luminal domain containing two potential N-linked glycosylation sites. D4ST-1 has significant amino acid identity with HNK-1 sulfotransferase (21.4%), N-acetylgalactosamine-4-O-sulfotransferase 1 (GalNAc-4-ST1) (24.7%), N-acetylgalactosamine-4-O-sulfotransferase 2 (GalNAc-4-ST2) (21.0%), chondroitin-4-O-sulfotransferase 1 (27.3%), and chondroitin-4-O-sulfotransferase 2 (22.8%). D4ST-1 transfers sulfate to the C-4 hydroxyl of beta1,4-linked GalNAc that is substituted with an alpha-linked iduronic acid (IdoUA) at the C-3 hydroxyl. D4ST-1 shows a strong preference in vitro for sulfate transfer to IdoUAalpha1,3GalNAcbeta1,4 that is flanked by GlcUAbeta1,3GalNAcbeta1,4 as compared with IdoUAalpha1,3GalNAcbeta1,4 flanked by IdoUAalpha1,3GalNAcbeta1,4. The specificity of D4ST-1 when assayed in vitro suggests that the addition of sulfate to GalNAc occurs immediately after epimerization of GlcUA to IdoUA. The open reading frame of D4ST-1 is encoded by a single exon located on human chromosome 15q14. Northern blot analysis reveals a single 2.4-kilobase transcript. D4ST-1 message is expressed in virtually all tissues at some level but is most highly expressed in pituitary, placenta, uterus, and thyroid. The properties of D4ST-1 indicate that sulfation of the GalNAc moieties in dermatan is mediated by a distinct GalNAc-4-O-sulfotransferase and occurs following epimerization of GlcUA to IdoUA.

Amino Acid Sequence↗

Molecular cloning and expression of an N-acetylgalactosamine-4-O-sulfotransferase that transfers sulfate to terminal and non-terminal beta 1,4-linked N-acetylgalactosamine.

We have identified and characterized an N-acetylgalactosamine-4-O-sulfotransferase designated GalNAc-4-ST2 (GenBank(TM) accession number ) based on its homology to HNK-1 sulfotransferase (HNK-1 ST). The cDNA predicts an open reading frame encoding a type II membrane protein of 443 amino acids with a 12-amino acid cytoplasmic domain, a 23-amino acid transmembrane domain, and a 408-amino acid luminal domain containing four potential N-linked glycosylation sites. GalNAc-4-ST2 displays a high degree of amino acid sequence identity with GalNAc-4-ST1 (46%), HNK-1 ST (23%), chondroitin 4-O-sulfotransferase-1 (C4ST-1) (27%), and chondroitin 4-O-sulfotransferase-2 (C4ST-2) (24%). GalNAc-4-ST2 transfers sulfate to the C-4 hydroxyl of terminal beta1,4-linked GalNAc in the sequence GalNAc-beta1,4GlcNAcbeta-R found on N-linked oligosaccharides and nonterminal beta1,4-linked GalNAc in chondroitin and dermatan. The translated region of GalNAc-4-ST2 is encoded by five exons located on human chromosome 18q11.2. Northern blot analysis reveals a 2.1-kilobase transcript. GalNAc-4-ST2 message is most highly expressed in trachea and to a lesser extent in heart, liver, pancreas, salivary gland, and testis. The I.M.A.G.E. cDNA clone 49547 contains a putative GalNAc-4-ST2 splice form with an open reading frame encoding a protein of 358 amino acids that lacks the transmembrane domain and the stem region. This form of GalNAc-4-ST2 is not retained by transfected cells and is active against chondroitin but not terminal beta1,4-linked GalNAc. Thus, as with GalNAc-4-ST1, sequences N-terminal to the catalytic domain contribute to the specificity of GalNAc-4-ST2 toward terminal beta1,4-linked GalNAc.

Acetylgalactosamine↗

Ifosfamide nephrotoxicity in pediatric cancer patients.

The renal functions in pediatric cancer patients who received ifosfamide (IFO) treatment were evaluated and the risk factors related to IFO nephrotoxicity were determined. The medical records of all children treated with IFO were reviewed, and 62 with normal renal function before IFO treatment were selected. Nephrotoxicity was diagnosed by measuring urine beta2-microglobulin and glucose, and serum phosphate, bicarbonate, and creatinine. Forty-eight (77.4%) had a history of previous cisplatin treatment. Nephrotoxicity was detected in 20 patients (32.3%). beta2-Microglobulinuria was observed in all 20, hypophosphatemia in 10 (16.1%), hypocarbia in 2 (3.2%), glucosuria in 5 (8.1%), and decreased creatinine clearance in 7 (11.3%). The cumulative dose of IFO and a history of previous cisplatin therapy were related to nephrotoxicity. Among the 20 patients with nephrotoxicity, the median cumulative dose of IFO in patients with a low (<500 mg/m2) and high (>500 mg/m2) cumulative dose of previous cisplatin was 80 g/m2 (73-102 g/m2) and 45 g/m2 (11-76 g/m2), respectively. Most of the nephrotoxicity persisted after cessation of IFO treatment. In conclusion, close monitoring of IFO nephrotoxicity should be started earlier in patients with high-dose cisplatin pretreatment. Tubular proteinuria, as indicated by beta2-microglobulinuria, was the most-sensitive marker for IFO nephrotoxicity. Long-term follow-up study for reversibility of IFO nephrotoxicity is in progress.

Adolescent↗

Moderate dietary fat consumption as a risk factor for ischemic heart disease in a population with a low fat intake: a case-control study in Korean men.

BACKGROUND: Dietary fat intake is associated with the incidence of ischemic heart disease (IHD) in Western countries. In populations in which both the average dietary fat consumption and the incidence of IHD are lower than in Western countries, the association of dietary fat intake with IHD incidence remains unknown. OBJECTIVE: We conducted a case-control study to examine the association of dietary fat with IHD incidence in Korean men. DESIGN: The case group consisted of 108 patients with electrocardiogram-confirmed myocardial infarction or angiographically confirmed (> or =50% stenosis) IHD who were admitted to a university teaching hospital in Seoul, Republic of Korea. The controls were 142 age-matched patients admitted to the departments of ophthalmology and orthopedic surgery at the same hospital. Dietary fat intake was assessed by a nutritionist using a semiquantitative food-frequency questionnaire. Body mass index (BMI), cigarette use, alcohol intake, exercise, and history of disease were determined during an interview and examination. RESULTS: In a univariate analysis, the mean percentages of energy from total fat, saturated fatty acids, and monounsaturated fatty acids were significantly higher in the cases than in the controls. BMI, smoking, and a history of hypertension were associated with the occurrence of IHD. In multiple logistic analyses, total fat intake was a significant risk factor (odds ratio: 1.08 for 1% of energy intake; 95% CI: 1.02, 1.14) after adjustment for BMI and smoking. CONCLUSION: In a population with a relatively low fat intake (19% of energy intake), a moderate increase in total fat intake may be a risk factor for IHD.

Alcohol Drinking↗

Application of progesterone measurement for age and body weight at puberty, and postpartum anestrus in Korean native cattle.

Studies were conducted in Korean native cattle to determine the age and body weight at puberty, and to determine the postpartum anestrus intervals based on plasma progesterone profiles, rectal palpation, ultrasonography and estrus observation. Korean native cattle reached puberty at 344 days of age and at 188 kg body weight as determined by plasma progesterone profiles, but estrous behavior was not detected until 3 months later, at a body weight of 215 kg. Prolonged postpartum anestrus occurred in 40% of smallholders cows, where animals were tethered and fed hay and rice straw, whereas its incidence was only 6.8% in a well managed herd that was grazed and housed. Plasma progesterone profiles were used to monitor ovarian activity postpartum in standard fed cows (n=44) and restricted fed cows (n=20). In standard fed cows 81.8% showed luteal activity by 50 days postpartum, compared to 55% in restricted fed cows. Approximately 40% of cows in both groups had shown observable estrous behavior by this time. This study indicates that one or more ovulations had occurred before pubertal heifers and postpartum cows showed overt estrous behavior. It was concluded that improved management and feeding practices should result in improved reproductive performance.

Age Factors↗

Inhibition of basal and stimulated progesterone synthesis by dichlorodiphenyldichloroethylene and methoxychlor in a stable pig granulosa cell line.

The effects of the insecticide dichlorodiphenyldichloroethylene (DDE) and methoxychlor in a stable pig granulosa cell line, JC-410, were investigated. The studies of DDE and methoxychlor were conducted in combination with studies of cholera toxin, the protein kinase A activator that stimulates cAMP and progesterone synthesis and gene expression of P450 cholesterol side chain cleavage (P450scc), which converts cholesterol to pregnenolone. Administration of DDE at 3000 and 10 000 ng ml (-1) was found to decrease progesterone synthesis 0.49- and 0.25-fold, respectively, and to block the stimulatory effect of 100 ng cholera toxin ml (-1), after 24 h incubation. At 1-100 ng ml (-1), methoxychlor did not affect progesterone synthesis after 48 h incubation. However, 1000 ng methoxychlor ml (-1) decreased progesterone synthesis 0.32-fold, and both 100 and 1000 ng methoxychlor ml (-1) blocked the stimulatory effect of cholera toxin. At 3000 and 10 000 ng ml(-1), DDE decreased cAMP synthesis 0.66-and 0.36-fold, respectively. At 300, 3000 and 10 000 ng ml (-1), DDE also decreased cholera toxin-stimulated cAMP synthesis 0.84-, 0.68-, and 0.52-fold, respectively. Administration of 1-100 ng methoxychlor ml (-1) did not affect basal or cholera toxin-stimulated cAMP synthesis. Cholera toxin increased P450scc mRNA 1.4-fold after 24 h incubation, while 3000 and 10 000 ng DDE ml (-1) led to 0.39- and 0.18-fold reductions, respectively. The stimulatory effect of cholera toxin on P450scc mRNA was blocked by 3000 and 10 000 ng DDE ml(-1). Cholera toxin increased P450scc mRNA 3.48-fold after 48 h incubation, while 100 and 1000 ng methoxychlor ml (-1) increased P450scc mRNA 1.79- and 3.0-fold, respectively, and further increased the stimulatory effect of cholera toxin 6.47- and 5.44-fold, respectively. The results of the present study indicate that DDE inhibits granulosa cell steroidogenesis by affecting cAMP production and P450scc gene expression. However, methoxychlor appears to inhibit steroidogenesis by a mechanism occurring before the conversion of cholesterol into pregnenolone.

Animals↗

Molecular cloning and expression of the pituitary glycoprotein hormone N-acetylgalactosamine-4-O-sulfotransferase.

N-Linked oligosaccharides terminating with the sequence SO(4)-4-GalNAcbeta1,4GlcNAcbeta1,2Manalpha are present on the pituitary hormones lutropin (LH), thyrotropin, and pro-opiomelanocortin. The sulfated structures on LH are essential for expression of its biologic function in vivo. We have cloned the N-acetylgalactosamine-4-sulfotransferase (GalNAc-4-ST1, GenBank(TM) accession number ), which mediates sulfate addition to the N-linked oligosaccharides on LH and other pituitary glycoproteins with terminal (beta1,4-linked GalNAc based on its homology to HNK-1 sulfotransferase (HNK-1 ST). GalNAc-4-ST1 displays 23% identity to HNK-1 ST and 28% to chondroitin 4-sulfotransferase 1 (C4ST-1) and 26% to chondroitin 4-sulfotransferase 2 (C4ST-2). The cDNA predicts a type II transmembrane protein of 424 amino acids with four potential N-linked glycosylation sites and a single membrane-spanning domain. GalNAc-4-ST1 has putative 5'-phosphosulfonate and 3'-phosphate binding sites. Three more carboxyl-terminal regions of unknown function also show a high degree of identity with HNK-1 ST, C4ST-1, and C4ST-2. The membrane-bound form of GalNAc-4-ST1 transfers sulfate to GalNAcbeta1, 4GlcNAcbeta-R but not to chondroitin, whereas truncated forms of GalNAc-4-ST1 that are released into the medium transfer sulfate to both GalNAcbeta1,4GlcNAcbeta-R and chondroitin. The first 118 amino acids of GalNAc-4-ST1 appear to contribute to both its activity and specificity for terminal beta1,4-linked GalNAc. GalNAc-4-ST1 also efficiently transfers sulfate to N-linked oligosaccharides on native LH and other glycoproteins terminating with beta1,4-linked GalNAc. A single transcript of 2.4 kilobases is most highly expressed in the pituitary and other regions of the central nervous system. The GalNAc-4-ST1 gene is located on human chromosome 19q13.1.

Amino Acid Sequence↗

Different stimulatory opioid effects on intracellular Ca(2+) in SH-SY5Y cells.

Present study revealed the stimulatory effects of delta opioid receptor on intracellular Ca(2+) concentration ([Ca(2+)](i)) in SH-SY5Y cells. Fura-2 based single cell fluorescence ratio (F345/F380) was used to monitor the fluctuation of [Ca(2+)](i). Application of the selective delta-opioid receptor agonist alone, [D-Pen(2,5)]-enkephalin (DPDPE), hardly had any effects on cells cultivated for 3-10 days. However, after the cells had been pre-stimulated with cholinoceptor agonist, carbachol, variable calcium elevation was found in 59% of the cultures. The response was naltridole-reversible and dose-dependent, and was abolished completely by thapsigargin (TG) treatment but not by administration of CdCl(2) or 0-Ca(2+) bath solutions. DPDPE-mediated [Ca(2+)](i) elevation was abolished by pertussis toxin (PTX) pretreatment but not cholera toxin (CTX), indicating coupling via G proteins of G(i)/G(o) subfamily. In 17.5% of the responding cells, biphase response was found which may be due to both the stimulatory and the inhibitory effects of opioid. On the other hand, in acutely dissociated cells, DPPDE alone induced [Ca(2+)](i) increase in 50% of the cultures. The probability and the amplitude of the elevation were decreased considerably by application of nifedipine or 0-Ca(2+) bath solution and was little affected by application of TG. DPDPE activated [Ca(2+)](i) increase via a PTX-insensitive and CTX-sensitive pathway suggesting coupling through G(s) subunit. All these indicated the opioid modulated the intracellular Ca(2+) regulation system through different pathways. SH-SY5Y cell line might be a suitable model for the investigation of the complex mechanism which underlies opioid function.

Analgesics, Opioid↗

Characterization of salicylic acid-responsive, arabidopsis Dof domain proteins: overexpression of OBP3 leads to growth defects.

Dof proteins are unique to plants and contain a single zinc finger DNA-binding domain called the Dof domain. OBP1, an Arabidopsis Dof protein, was previously isolated through an interaction with OBF4, an ocs element-binding protein. Two additional Dof proteins, called OBP2 and OBP3, were isolated through homologous screening. All three OBP proteins contain transcriptional activation domains in their C-terminal region. While no significant differences were found between the OBP proteins in terms of their DNA and protein-binding properties, tissue-specific RNA expression patterns were found. The RNA expression levels of all three OBP proteins increased following treatment with auxin, salicylic acid (SA) or cycloheximide, although the level of induction varied among the different proteins and in the tissues tested. These results suggest that even though they have similar DNA binding and protein-protein interaction properties, the different OBP proteins are likely to have distinct functions in specific parts of the plant. There is a good correlation between the expression of the OBP proteins and the ocs element, a stress-response element which is also induced by auxin, SA and cycloheximide. To begin to analyze the function of the OBP proteins, transgenic lines overexpressing OBP3 were generated. These plants showed a severe growth defect with altered root development and yellowish leaves. The severity of the growth defects correlated with OBP3 expression levels and in some cases led to death, suggesting that some Dof proteins play important roles in plant growth and development.

Amino Acid Sequence↗

[Real-time techniques in monitoring secretion activity in cells].

Chemical communication for neurons and endocrine cells depends on their secretion activity. Development of variety of biophysical techniques have greatly promoted the understanding of the mechanism of cellular secretion. Here we will introduce three recently developed biophysical techniques which have been frequently used to monitor secretion activity in real-time.

Cells↗

[Cellular electrophysiology and patch clamp technique].

The fundamentals of cellular electrophysiology and its basic method are described here. The simplyfied structure of the cell, the chemical constitutes of the membrane, and the mechanism of the bioelectric signal generation are included. The patch clamp is the fundamental methodology in modern electrophysiology research. It is closely related to the construction of the laboratory setup and the key technique in experimental procedure.

Cell Membrane↗

[Single channel and whole cell recordings using patch clamp technique].

Action of ion channel on membrane is the key events of messenger transduction for excitable cells, which can be detected by the patch clamp technique. The developments of patch clamp technique have brought a revolution of life science research. The theory, work modes, single channel and whole cell recording techniques in single cells are described here in detail.

Cells, Cultured↗

Identification of a gene, SOO1, which complements osmo-sensitivity and defect in in vitro beta1,3-glucan synthase activity in Saccharomyces cerevisiae.

The soo1-1 (for suppressor of osmo-sensitivity) mutation results in a temperature-dependent osmo-sensitive phenotype at non-permissive temperature (37 degrees C), makes yeast cells more susceptible to Zymolyase and affects in vitro beta1,3-glucan synthase activity level. Transformation of soo1-1 mutant by SOO1 gene reverses these effects. Nucleotide sequencing of SOO1 revealed that this gene is identical to the recently reported alpha-COP that is involved in the intracellular protein translocation from endoplasmic reticulum to Golgi and vice versa. Although the soo1-1 mutant strain showed osmotically remediable growth at 37 degrees C, SOO1 disruptants could not grow in any culture conditions. Analysis of cell wall fractions revealed that the soo1-1 mutation causes a decrease in in vitro beta1,3-glucan synthase activity, and, thus, a subsequent alteration of the cell wall composition.

Cell Wall↗

Laparoscopic tubal anastomosis: fertility outcome in 202 cases.

OBJECTIVE: To evaluate the fertility outcome after laparoscopic tubal anastomosis for reversal of sterilization. DESIGN: Retrospective clinical study. SETTING: A private practice affiliated with a university medical school. PATIENT(S): Two hundred two women who desired reversal of tubal sterilization. INTERVENTION(S): Laparoscopic tubal anastomosis. MAIN OUTCOME MEASURE(S): The cumulative pregnancy rate (PR) and factors that influenced the fertility outcome. RESULT(S): The cumulative PR in the 186 patients for whom follow-up data were available was 60.3%, 79.4%, and 83.3% at 6, 12, and 18 months after operation, respectively. Five patients (3.2%) had ectopic pregnancies; one of these patients subsequently conceived normally. There were no statistically significant differences in the PR according to the sterilization method used, the site of the tubal anastomosis, or the length of the fallopian tube after surgery. The intrauterine PR was 87.1% (149/171) with bilateral anastomosis and 60% (9/15) with unilateral anastomosis. The PR decreased with increasing patient age (mean [+/- SD], 35+/-3.6 years) but was still 70.6% (12/17) in patients aged 40-45 years. CONCLUSION(S): Our findings suggest that laparoscopic tubal anastomosis is a highly successful procedure. This less invasive approach could be considered the procedure of choice in patients who desire reversal of tubal sterilization.

Adult↗

Identification of a rice APETALA3 homologue by yeast two-hybrid screening.

A cDNA clone OsMADS16 was isolated from the rice young inflorescence cDNA expression library by the yeast two-hybrid screening method with OsMADS4 as bait. We have previously shown that the OsMADS4 gene is a member of the PI family and that the MADS-box gene is involved in controlling development of the second and third whorls of rice flowers. The sequence comparison indicated that OsMADS16 belongs to the AP3 family. The OsMADS16 protein contains a PI-derived motif, FAFRVVPSQPNLH, that is a conserved sequence in AP3 family genes at the C-terminal region. In addition, OsMADS16 contains a paleoAP3 motif, YGGNHDLRLG, downstream of the PI-derived motif. The paleoAP3 motif is a consensus sequence in the C-terminal region of the AP3 family genes of lower eudicot and magnolid dicot species. RNA blot analysis showed that the OsMADS16 gene was expressed in the second and third whorls, whereas the OsMADS4 transcripts were present in the second, third, and fourth whorls. These expression patterns of the OsMADS16 and OsMADS4 genes are very similar to those of AP3 and PI, respectively. In the yeast two-hybrid system, OsMADS4 interacted only with OsMADS16 among several rice MADS genes investigated, suggesting that OsMADS4 and OsMADS16 function as a heterodimer in specifying sepal and petal identities. The OsMADS16 protein displayed transcription activation ability in yeast, whereas AP3 did not. It was also shown in yeast that OsMADS16 interacted with PI whereas OsMADS4 did not interact with AP3. These differences between OsMADS16 and AP3 indicate that the functions of the AP3 family genes of monocots and dicots diverged during molecular evolution processes of the B function genes. Deletion analysis showed that the 155-200 amino acid region of the OsMADS16 protein plays an important role in the transcription activation ability.

Amino Acid Sequence↗

A glucocorticoid-inducible transcription system causes severe growth defects in Arabidopsis and induces defense-related genes.

A glucocorticoid-inducible transcription system was employed to control the expression of AtEBP, an Arabidopsis transcription factor. A number of the transgenic AtEBP lines had developmental and growth defects when grown on dexamethasone (DEX), a strong synthetic glucocorticoid. However, these growth defects were not confined to the AtEBP lines but were observed with other transgenic lines that were generated using the same system, including empty vector lines. In about 25% of the AtEBP or empty vector transgenic lines, these growth defects were severe and in some cases led to death. As AtEBP has been linked to the plant defense response, the expression of specific defense-related genes, including a number of pathogenesis-related (PR) genes was also examined. PDF1.2, a plant defensin gene, was strongly induced in all transgenic lines examined following treatment with DEX, including empty vector lines that did not show any observable DEX-induced growth defect. PR-5 was induced to a lesser extent in all the lines, while the expression of PR-1, PR-2 and phenylalanine ammonia-lyase 3 (PAL3) did not change significantly. While the induction of the AtEBP transgene and PDF1.2 had similar DEX concentration requirements, the kinetics of induction differed significantly, with the AtEBP transgene being induced within 1 h and PDF1.2 only being induced between 24 and 48 h. Although the molecular mechanisms underlying the growth defects and changes in gene expression remain to be determined, these changes appear to result from the glucocorticoid-inducible system itself, and may therefore limit the usefulness of this system for controlling gene expression in Arabidopsis.

Arabidopsis↗

Determination of the motif responsible for interaction between the rice APETALA1/AGAMOUS-LIKE9 family proteins using a yeast two-hybrid system.

A MADS family gene, OsMADS6, was isolated from a rice (Oryza sativa L.) young flower cDNA library using OsAMDS1 as a probe. With this clone, various MADS box genes that encode for protein-to-protein interaction partners of the OsMADS6 protein were isolated by the yeast two-hybrid screening method. On the basis of sequence homology, OsMADS6 and the selected partners can be classified in the APETALA1/AGAMOUS-LIKE9 (AP1/AGL9) family. One of the interaction partners, OsMADS14, was selected for further study. Both genes began expression at early stages of flower development, and their expression was extended into the later stages. In mature flowers the OsMADS6 transcript was detectable in lodicules and also weakly in sterile lemmas and carpels, whereas the OsMADS14 transcript was detectable in sterile lemmas, paleas/lemmas, stamens, and carpels. Using the yeast two-hybrid system, we demonstrated that the region containing of the 109th to 137th amino acid residues of OsMADS6 is indispensable in the interaction with OsMADS14. Site-directed mutation analysis revealed that the four periodical leucine residues within the region are essential for this interaction. Furthermore, it was shown that the 14 amino acid residues located immediately downstream of the K domain enhance the interaction, and that the two leucine residues within this region play an important role in that enhancement.

Amino Acid Sequence↗