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Biomedical subjects

M J Kim

Publications and source records attributed to M J Kim.

At least 163 records · Page 9Linked to original sources

Overexpression of phospholipase C-gamma1 suppresses UVC-induced apoptosis through inhibition of c-fos accumulation and c-Jun N-terminal kinase activation in PC12 cells.

Ultraviolet-C (UVC) irradiation induces DNA damage and UVC-irradiated cells undergo cell growth arrest to repair the damaged DNA or the induction of apoptosis to prevent the risk of neoplastic transformation. Phospholipase C-gamma1 (PLC-gamma1) is a mediator of growth factor induced-signal cascade, catalyzing the hydrolysis of phosphatidyl 4,5-bisphosphate to generate second messengers, diacylglycerol and inositol 1,4,5-trisphosphate (IP(3)). PLC-gamma1 is activated by phosphorylation of tyrosine residues upon occupation of cell surface receptors by growth factors and plays an important role in controlling cellular proliferation and differentiation. In this study, we found that PLC-gamma1 was tyrosine phosphorylated within 2.5 min after UVC irradiation. To investigate the role of UVC-induced tyrosine phosphorylation of PLC-gamma1, we compared the effect of UVC between PLC-gamma1 overexpressing cells and empty vector transfected cells. Overexpression of PLC-gamma1 inhibited UVC-induced sub-diploid peak and DNA fragmentation. Northern blot analysis revealed that UVC-induced c-fos mRNA accumulation was inhibited in PLC-gamma1 overexpressing cells, while c-jun expression was not affected. In addition, UVC-induced activation of c-Jun N-terminal kinase (JNK) was significantly suppressed in PLC-gamma1 overexpressing cells. These results suggest that PLC-gamma1 may associate with the protective function against the UVC-induced cell death progression via the inhibition of accumulation of c-fos mRNA and the inhibition of JNK kinase activity.

Animals↗

Crystal structure of a maltogenic amylase provides insights into a catalytic versatility.

Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. Compared with many different amylases that are able to hydrolyze only alpha-D-(1,4)-glycosidic bonds, maltogenic amylases exhibit catalytic versatility: hydrolysis of alpha-D-(1,4)- and alpha-D-(1,6)-glycosidic bonds and transglycosylation of oligosaccharides to C3-, C4-, or C6-hydroxyl groups of various acceptor mono- or disaccharides. It has been speculated that the catalytic property of the enzymes is linked to the additional approximately 130 residues at the N terminus that are absent in other typical alpha-amylases. The crystal structure of a maltogenic amylase from a Thermus strain was determined at 2.8 A. The structure, an analytical centrifugation, and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N-terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (alpha/beta)(8) barrel of the adjacent subunit. The active site is a narrow and deep cleft suitable for binding cyclodextrins, which are the preferred substrates to other starch materials. At the bottom of the active site cleft, an extra space, absent in the other typical alpha-amylases, is present whose size is comparable with that of a disaccharide. The space is most likely to host an acceptor molecule for the transglycosylation and to allow binding of a branched oligosaccharide for hydrolysis of alpha-D-(1,4)-glycosidic or alpha-D-(1,6)-glycosidic bond. The (alpha/beta)(8) barrel of the enzyme is the preserved scaffold in all the known amylases. The structure represents a novel example of how an enzyme acquires a different substrate profile and a catalytic versatility from a common active site and represents a framework for explaining the catalytic activities of transglycosylation and hydrolysis of alpha-D-(1,6)-glycosidic bond.

Amino Acid Sequence↗

Mobile gene cassettes and integrons in evolution.

Integrons and the site-specific recombination systems encoded by them provide a simple mechanism for the addition of new genes to bacterial chromosomes. Although there is substantial divergence among the four known integron-encoded integrases, they all recognize the recombination sites, known as 59-base elements, that are associated with genes that are packaged in gene cassettes. In contrast, the integron-associated recombination sites, attl sites, are preferentially recognized by the cognate integrase.

Base Sequence↗

Genetic dissection of protein-protein interactions in multi-tRNA synthetase complex.

Cytoplasmic aminoacyl-tRNA synthetases of higher eukaryotes acquired extra peptides in the course of their evolution. It has been thought that these appendices are related to the occurrence of the multiprotein complex consisting of at least eight different tRNA synthetase polypeptides. This complex is believed to be a signature feature of metazoans. In this study, we used multiple sequence alignments to infer the locations of the peptide appendices from human cytoplasmic tRNA synthetases found in the multisynthetase complex. The selected peptide appendices ranged from 22 aa of aspartyl-tRNA synthetase to 267 aa of methionyl-tRNA synthetase. We then made genetic constructions to investigate interactions between all 64 combinations of these peptides that were individually fused to nonsynthetase test proteins. The analyses identified 11 (10 heterologous and 1 homologous) interactions. The six peptide-dependent interactions paralleled what had been detected by crosslinking methods applied to the isolated multisynthetase complex. Thus, small peptide appendices seem to link together different synthetases into a complex. In addition, five interacting pairs that had not been detected previously were suggested from the observed peptide-dependent complexes.

Amino Acid Sequence↗

The ginsenoside Rg3 evokes endothelium-independent relaxation in rat aortic rings: role of K+ channels.

The purpose of the present study was to characterize the mechanism underlying the direct relaxing activity of ginsenosides on vascular smooth muscle. The total ginsenoside mixture, ginsenosides from either the protopanaxadiol group or the protopanaxatriol group, and the ginsenoside Rg3 from the protopanaxatriol group caused a concentration-dependent relaxation of rat aortic rings without endothelium contracted with 25 x 10(-3) M KCl but affected only minimally those contracted with 60 x 10(-3) M KCl. Ginsenoside Rg3 was the most potent relaxing agonist. Relaxations elicited by ginsenoside Rg3 were markedly reduced by tetraethylammonium, a blocker of non-selective K+ channels, but not by glibenclamide, a blocker of ATP-sensitive K+ channels. Ginsenoside Rg3 significantly inhibited Ca2+-induced concentration-contraction curves and the 45Ca2+ influx in aortic rings incubated with 25 x 10(-3) M KCl whereas these responses were not affected in rings incubated with 60 x 10(-3) M KCl. Ginsenoside Rg3 caused a time- and concentration-dependent efflux of 86Rb from aortic rings that was inhibited by tetraethylammonium but not by glibenclamide. These findings indicate that ginsenoside Rg3 is a potent inhibitor of vascular smooth muscle tone and that this effect seems to be due to an inhibition of Ca2+ influx and stimulation of K+ efflux, possibly via activation of tetraethylammonium-sensitive K+ channels.

Animals↗

Comparative study of transrectal ultrasonography, pelvic computerized tomography, and magnetic resonance imaging in preoperative staging of rectal cancer.

PURPOSE: The preoperative assessment of rectal cancer wall invasion and regional lymph node metastasis is essential for the planning of optimal therapy. This study was done to determine the accuracy and clinical usefulness of transrectal ultrasonography, pelvic computed tomography, and magnetic resonance imaging in preoperative staging. METHODS: A total of 89 patients with rectal cancer were examined with transrectal ultrasonography (n = 89), pelvic computed tomography (n = 69), and magnetic resonance imaging with endorectal coil (n = 73). The results obtained by these diagnostic modalities were compared with the histopathologic staging of specimens. RESULTS: In staging depth of invasion, the overall accuracy was 81.1 percent (72/89) by transrectal ultrasonography, 65.2 percent (45/ 69) by computed tomography, and 81 percent (59/73) by magnetic resonance imaging. Overstaging was 10 percent (9/89) by transrectal ultrasonography, 17.4 percent (12/69) by computed tomography, and 11 percent (8/73) by magnetic resonance imaging; and understaging was 8 of 89 (8.9 percent) by transrectal ultrasonography, 12 of 69 (17.4 percent) by computed tomography, and 6 of 73 (8 percent) by magnetic resonance imaging. In staging lymph node metastasis, the overall accuracy rate was 54 of 85 (63.5 percent) in transrectal ultrasonography, 39 of 69 (56.5 percent) in computed tomography, and 46 of 73 (63 percent) in magnetic resonance imaging. The sensitivity was 24 of 45 (53.3 percent) in transrectal ultrasonography, 14 of 25 (56 percent) in computed tomography, and 33 of 42 (78.5 percent) in magnetic resonance imaging; and specificity was 30 of 40 (75.0 percent) in transrectal ultrasonography, 25 of 44 (56.8 percent) in computed tomography, and 13 of 31 (41.9 percent) in magnetic resonance imaging. The accuracy in detection of positive lateral pelvic lymph nodes under magnetic resonance imaging (n = 8) was 12.5 percent. The accuracy in detection of posterior vaginal wall invasion was 100 percent in transrectal ultrasonography (n = 7) and 100 percent in magnetic resonance imaging (n = 3), but 28.5 percent in computed tomography (n = 7). CONCLUSIONS: Both transrectal ultrasonography and magnetic resonance imaging with endorectal coil exhibited similar accuracy and were superior to conventional computed tomography in preoperative assessment of depth of invasion and adjacent organ invasion. Because transrectal ultrasonography is a safer and more cost-effective modality than magnetic resonance imaging, transrectal ultrasonography is an appropriate method for preoperative staging of rectal cancer. Further efforts will be needed to provide a better staging of lymph node involvement.

Adult↗

Cytotoxicity of urushiols isolated from sap of Korean lacquer tree (Rhus vernicifera Stokes).

Cytotoxicities of four urushiols, congeners isolated from the sap of Korean lacquer tree (Rhus vernicifera Stokes), to 29 human cancer cell lines originated from 9 organs were evaluated. Their values of 50% growth inhibition were below 4 microg/ml, and showed cell line specific cytotoxicity. The present result is the first report on the cytotoxicity of urushiols suggesting that they would have an anticancer activity to human cancer cells.

Antineoplastic Agents↗

Posterior capsule opacification in eyes with a silicone or poly(methyl methacrylate) intraocular lens.

PURPOSE: To evaluate the effect of poly(methyl methacrylate) (PMMA) and silicone intraocular lenses (IOLs) on posterior capsule opacification (PCO) after cataract surgery. SETTING: Kangnam St. Mary's Hospital, Seoul, Korea. METHODS: This retrospective study comprised 48 patients (54 eyes) who had neodymium:YAG (Nd:YAG) laser posterior capsulotomy from March 1995 to December 1997. All operations were performed by 1 surgeon using the same technique except for incision method. RESULTS: Mean interval from cataract surgery to Nd:YAG capsulotomy was 31 months in the PMMA group and 15 months in the silicone group. The difference between groups was statistically significant (P = .0002). The ratio of Elschnig pearl to fibrosis type PCO was 16:6 in the PMMA group and 14:18 in the silicone group. Mean total Nd:YAG laser energy used was 256 mJ in the PMMA group and 309 mJ in silicone group. However, the damage caused by the laser was more severe and more common in the silicone group. CONCLUSION: Silicone IOLs induced PCO faster than PMMA IOLs, with fibrosis the most common type in the silicone group. Precautions should be taken to prevent damage during Nd:YAG laser capsulotomy in eyes with a silicone IOL.

Cataract↗

The involvement of cAMP signaling pathway in axis specification in Xenopus embryos.

The cAMP signaling system has been postulated to be involved in embryogenesis of many animal species, however, little is known about its role in embryonic axis formation in vertebrates. In this study, the role of the cAMP signaling pathway in patterning the body plan of the Xenopus embryo was investigated by expressing and activating the exogenous human 5-hydroxytryptamine type 1a receptor (5-HT(1a)R) which inhibits adenylyl cyclase through inhibitory G-protein in embryos in a spatially- and temporally-controlled manner. In embryos, ventral, but not dorsal expression and stimulation of this receptor during blastula and gastrula stages induced secondary axes but were lacking anterior structures. At the molecular level, 5-HT(1a)R stimulation induced expression of the dorsal mesoderm marker genes, and downregulated expression of the ventral markers but had no effect on expression of the pan mesodermal marker gene in ventral marginal zone explants. In addition, ventral expression and stimulation of the receptor partially restored dorsal axis of UV-irradiated axis deficient embryo. Finally, the total mass of cAMP differs between dorsal and ventral regions of blastula and gastrula embryos and this is regulated in a temporally-specific manner. These results suggest that the cAMP signaling system may be involved in the transduction of ventral signals in patterning early embryos.

Animals↗

Transglycosylation of naringin by Bacillus stearothermophilusMaltogenic amylase to give glycosylated naringin.

Naringin, a bitter compound in citrus fruits, was transglycosylated by Bacillus stearothermophilus maltogenic amylase reaction with maltotriose to give a series of mono-, di-, and triglycosylnaringins. Glycosylation products of naringin were observed by TLC and HPLC. The major glycosylation product was purified by using a Sephadex LH-20 column. The sturcture was determined by using MALDI-TOF MS, methylation analysis, and (1)H and (13)C NMR. The major transglycosylation product was maltosylnaringin, in which the maltose unit was attached by an alpha-1-->6 glycosidic linkage to the D-glucose moiety of naringin. This product was 250 times more soluble in water and 10 times less bitter than naringin.

Antioxidants↗

Prophylactic efficacy of lithium administered every second day: a WHO multicentre study.

OBJECTIVES: To study the prophylactic efficacy of lithium administered every second day to patients with bipolar disorder or recurrent unipolar depressive disorder. METHODS: The study was carried out as a WHO multicentre study in five different psychiatric clinics: Russia (Moscow), Canada (Montreal), India (Lucknow), Germany (Munich) and South Korea (Pusan), with the lithium tablets being supplied from Denmark (Copenhagen). Participation in the study was conditional on the patient having been in prophylactic lithium treatment for the preceding 2-year period and having been free of depressive or manic phases during the preceding 6 months. After a 2-month baseline period during which lithium was administered every day, the treatment was changed to lithium intake every second day, this regimen in most cases being continued for 22 months. RESULTS: Forty-seven patients, 42 with a diagnosis of bipolar disorder and five with a diagnosis of recurrent unipolar depressive disorder, participated in the study. The number of patients from each centre ranged from six to 11. The mean lithium dose every second day was 36 mmol lithium, leading to a mean 12-h standard serum lithium concentration during the last month of treatment at 0.78+/-0.16 mmol/L. Lithium intake every second day was effective in 38 out of 47 patients. The remaining nine patients had recurrences, a number not differing from the 16 patients in the same group of 47 patients who, during the 2-year period preceding the study, had recurrences. The mean 12-h standard serum lithium concentration was lower in the patients who had recurrences than in those who stayed euthymic. The results suggest that to be effective when administered every second day, lithium must be given in a dose leading to a 12-h standard serum lithium concentration at 0.8 mmol/L or above. CONCLUSIONS: In the dose administered, lithium intake every second day was as effective as lithium intake every day. The amount and severity of lithium-related side effects were apparently not influenced by the change to lithium intake every second day.

Adult↗

Heat shock response decreases endotoxin-induced acute lung injury in rats.

OBJECTIVE: Transient whole-body hyperthermia was reported to reduce lung damage in a rat with intra-abdominal sepsis produced by caecal perforation. METHODOLOGY: In order to determine the effect of heat shock response on acute lung injury induced by endotoxin, which plays a central role in the pathogenesis of sepsis, we instilled either saline or lipopolysaccharide (LPS) intravenously with and without heat pretreatment in rats. The heated rats had their rectal temperature raised to more than 40 degrees C for 13 min 18 h before intravenous administration of saline or LPS. RESULTS: We found that the lung leak was significantly increased among the rats given LPS intravenously with (median, 0.17; range, 0.15-0.22; n = 10) and without heat pretreatment (0.23; 0.17-0.30; n = 10) compared with those of saline-treated rats (0.13; 0.10-0.14; n = 10) (P < 0.05 in each). However, rats given LPS after heat pretreatment had significantly decreased lung leak index compared with those of LPS-treated rats without heat pretreatment (P < 0.05). Rats administered LPS intravenously showed increased myeloperoxidase activity without heat pretreatment (19.01; 9.34-28.00 U/g; n = 10) compared with that of saline-treated rats (7.09; 4.49-10.56 U/g; n = 5) (P < 0.05) (Fig. 2). Myeloperoxidase activity of the rats treated with LPS with heat pretreatment (5.57; 2.87-8.96 U/g; n = 10) was significantly decreased to the level of normal control compared with that of LPS-treated rats without heat pretreatment (P < 0.05). The levels of heat shock proteins (HSP72) in lung tissue, which were examined by western blot analysis, were increased over baseline levels at 23 h after hyperthermic stress. CONCLUSIONS: These observations show that brief heat shock response is associated with the induction of HSP72 protein synthesis and attenuated neutrophil recruitment and acute lung leak is induced by endotoxin in rats.

Acute Disease↗

Molecular cloning of multiple splicing variants of JIP-1 preferentially expressed in brain.

Stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) is activated by a variety of cellular or environmental stresses. Proper regulation of the SAPK/JNK pathway may be critical for cell survival or death under various conditions. In this study, we report the molecular cloning of novel isoforms of JIP-1, which harbor a putative phosphotyrosine interaction domain and a helix-loop-helix domain, as well as an SH3 homologous region in the C terminus. Northern analysis indicates that transcription variant jip-1 is expressed in brain and kidney and transcription variants jip-2 and jip-3 are specifically expressed in brain. In situ hybridization data showed that the hybridized jip messages were heavily concentrated in adult brain, and were particularly enriched in the cerebral cortex and hippocampus, the brain regions vulnerable to pathological states such as hypoxia-ischemia, epilepsy, and Alzheimer's disease. All the deduced protein products of the jip transcription variants appear to have a similar property in that they inhibit the SAPK/JNK stimulation when overexpressed. Inhibition of SAPK activation by overexpression of the novel isoform JIP-2a resulted in suppression of etoposide-induced cell death in a neuroglioma cell line, N18TG. These findings suggest that JIP may play an important role in regulation of the SAPK pathway that is involved in stress-induced cellular responses.

Adaptor Proteins, Signal Transducing↗

Response to chloroquine of Plasmodium vivax among South Korean soldiers.

The response to standard chloroquine treatment was evaluated, by microscopical examination of blood-smears, among 81 soldiers diagnosed with Plasmodium vivax malaria in South Korea in 1996. The smears were prepared pre-treatment and 3, 14 and 28 days after starting chemotherapy. Parasitaemias were determined after staining the smears with Giemsa's stain. Blood samples from the patients who were not smear-negative by day 3 were carefully checked for parasites, by staining smears with Acridine Orange and by a PCR-based assay. Only two of the patients appeared to be parasitaemic on day 14 and were therefore considered treatment failures. Although both were apparently cured after additional therapy with the same regimen, one had a recurrence 8 months later. Most cases of recent, resurgent malaria in South Korea therefore appear to sensitive to chloroquine.

Adult↗

Expression and immunologic characterization of recombinant heat shock protein 58 of Leptospira species: a major target antigen of the humoral immune response.

A clone of Leptospira interrogans serovar lai that was isolated by immunoscreening of a genomic lambda library with sera from convalescent patients with leptospirosis directed expression of a unique 62-kDa protein in Escherichia coli. When examined by SDS-PAGE, the protein comigrated with an immunodominant protein present in leptospiral cell lysate. Determination of the nucleotide sequence of the 2.7-kb insert DNA identified two genes homologous to the hsp58 and hsp10 of L. interrogans serovar copenhageni reported previously. The overexpressed recombinant Hsp58 protein was purified and used to immunize a rabbit to produce a polyclonal antibody. Immunoblot analysis using the rabbit anti-Hsp58 G antibodies showed that the 62-kDa protein was commonly present in lysates of other serovars of leptospires, consistent with the strong sequence conservation between the hsp58 genes of the two serovars. Immunoglobulin G antibodies to the Hsp58 were specifically detected by ELISA in 82% of sera (18/22) from patients with leptospirosis. Deletion analysis of the recombinant Hsp58 protein indicated that a strong antigenic determinant for humoral immune response is located between amino acids 360 and 380 (DREKLQERLAKLAGGVAVIHV) of Hsp58, which are highly conserved among the GroEL family. The strong sequence conservation of the Hsp58 among leptospires and its importance as a major target for the humoral immune response warrant further studies of its potential pathogenetic role.

Animals↗

Modes of action of acarbose hydrolysis and transglycosylation catalyzed by a thermostable maltogenic amylase, the gene for which was cloned from a Thermus strain.

A maltogenic amylase gene was cloned in Escherichia coli from a gram-negative thermophilic bacterium, Thermus strain IM6501. The gene encoded an enzyme (ThMA) with a molecular mass of 68 kDa which was expressed by the expression vector p6xHis119. The optimal temperature of ThMA was 60 degrees C, which was higher than those of other maltogenic amylases reported so far. Thermal inactivation kinetic analysis of ThMA indicated that it was stabilized in the presence of 10 mM EDTA. ThMA harbored both hydrolysis and transglycosylation activities. It hydrolyzed beta-cyclodextrin and starch mainly to maltose and pullulan to panose. ThMA not only hydrolyzed acarbose, an amylase inhibitor, to glucose and pseudotrisaccharide (PTS) but also transferred PTS to 17 sugar acceptors, including glucose, fructose, maltose, cellobiose, etc. Structural analysis of acarbose transfer products by using methylation, thin-layer chromatography, high-performance ion chromatography, and nuclear magnetic resonance indicated that PTS was transferred primarily to the C-6 of the acceptors and at lower degrees to the C-3 and/or C-4. The transglycosylation of sugar to methyl-alpha-D-glucopyranoside by forming an alpha-(1,3)-glycosidic linkage was demonstrated for the first time by using acarbose and ThMA. Kinetic analysis of the acarbose transfer products showed that the C-4 transfer product formed most rapidly but readily hydrolyzed, while the C-6 transfer product was stable and accumulated in the reaction mixture as the main product.

Acarbose↗

The Drosophila tissue polarity gene starry night encodes a member of the protocadherin family.

The tissue polarity genes control the polarity of hairs, bristles and ommatidia in the adult epidermis of Drosophila. We report here the identification of a new tissue polarity gene named starry night (stan). Mutations in this essential gene alter the polarity of cuticular structures in all regions of the adult body. The detailed polarity phenotype of stan on the wing suggested that it is most likely a component of the frizzled (fz) pathway. Consistent with this hypothesis, stan appears to be downstream of and required for fz function. We molecularly cloned stan and found that it encodes a huge protocadherin containing nine cadherin motifs, four EGF-like motifs, two laminin G motifs, and seven transmembrane domains. This suggests that Stan functions in signal reception, perhaps together with Fz.

Amino Acid Motifs↗