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Biomedical subjects

H K Kim

Publications and source records attributed to H K Kim.

At least 55 records · Page 3Linked to original sources

Significance of ecto-cyclase activity of CD38 in insulin secretion of mouse pancreatic islet cells.

Cyclic ADP-ribose (cADPR), a product of CD38, has a second messenger role for in intracellular Ca(2+) mobilization from microsomes of pancreatic islets as well as from a variety of other cells. ADP-ribosylation of CD38 by ecto-mono ADP-ribosyltransferase in activated T cells results in apoptosis as well as inactivation of its activities. We, therefore, examined the effect of ADP-ribosylation of CD38 in mouse pancreatic islet cells. NAD-dependent inactivation and ADP-ribosylation of CD38, intracellular concentrations of cADPR and Ca(2+), and insulin secretion were measured following incubation of mouse pancreatic islet cells with NAD. ADP-ribosylation of CD38 inactivated its ecto-enzyme activities, and abolished glucose-induced increase of cADPR production, intracellular concentration of Ca(2+), and insulin secretion. Taken together, ecto-cyclase activity of CD38 to produce intracellular cADPR seems to be indispensable for insulin secretion.

ADP-ribosyl Cyclase↗

Fungicidal effect of antimicrobial peptide, PMAP-23, isolated from porcine myeloid against Candida albicans.

The antifungal activity and mechanism of a 23-mer peptide, PMAP-23, derived from pig myeloid was investigated. PMAP-23 displayed strong antifungal activity against yeast and mold. To investigate the antifungal mechanism of PMAP-23, fluorescence activated flow cytometry and confocal laser scanning microscopy were performed. Candida albicans treated with PMAP-23 showed higher fluorescence intensity by propidium iodide(PI) staining, which was similar to that of Melittin than untreated cells. Confocal microscopy showed that the peptide was located in the plasma membrane. The action of peptides against fungal cell membranes was examined by treating prepared protoplasts of C. albicans with the peptide and lipid vesicle titration test. The result showed that the peptide prevented the regeneration of fungal cell walls and induced release of the fluorescent dye trapped in the artificial membrane vesicles, indicating that the peptide exerts its antifungal activity by acting on the plasma lipid membrane.

Amino Acid Sequence↗

Plasma fibronectin levels in ischemic heart disease.

Fibronectin is a paradigm adhesive protein which has been implicated in the regulation of several cellular processes and cell-cell interactions. Large amounts of fibronectin have been detected in atherosclerotic plaques, while hypertension in animal models has been shown to rapidly increase fibronectin expression in arterial walls. The aim of the present study was to determine the levels of plasma fibronectin (FN) in 133 patients with ischemic heart disease and in 36 normal controls, and to investigate the possible association with blood pressure. Plasma FN levels in patients with ischemic heart disease were found to be significantly elevated (mean+/-S.D.; 46.5+/-14.2 mg/dl) compared with the control group (38.0+/-14.2 mg/dl) (P=0.002). Plasma FN concentrations were significantly different between the hypertensive group (52.9+/-14.5 mg/dl) and the normal blood pressure group (41.4+/-11.8 mg/dl) among the patients with ischemic heart disease (P<0.001). Plasma FN concentration was positively correlated with total cholesterol, triglyceride, systolic blood pressure and body mass index. In conclusion, the plasma fibronectin level may have pathogenetic implications in association with lipid components and blood pressure in patients with ischemic heart disease.

Adult↗

Synthesis and biological properties of C-2, C-8, N-9 substituted 6-(3-chloroanilino)purine derivatives as cyclin-dependent kinase inhibitors. Part II.

In this study, C-2, C-8, N-9 substituted 6-(3-chloroanilino)purine derivatives were synthesized and their inhibitory effects on cyclin-dependent kinases (CDK2, 4) as well as their cytotoxicities were evaluated. The effects of substituents at the C-2, C-8, and N-9 positions of the substituted purine were investigated. Among the compounds tested, [6-(3-chloroanilino)-2-(2-hydroxymethyl-4-hydroxypyrrolidyl)-9- isopropylpurine] (4h) was the most active inhibitor of CDK2 with IC50 of 0.3 microM, i.e. a two-fold increased inhibitory activity as compared to roscovitine. Results from structure-activity relationship studies should allow the design of more potent and selective CDK2 inhibitors, which may provide an effective therapy for cancer or other CDK-dependant diseases.

Antineoplastic Agents↗

Determination of propranolol concentration in small volume of rat plasma by HPLC with fluorometric detection.

A simple, rapid and sensitive fluorescence high performance liquid chromatographic method was developed to determine propranolol concentration in the small volume of rat plasma without the solvent extraction step using pronethanol as the internal standard. The analysis was accomplished using a 5 microm CAPCELL PAK analytical cyano column at room temperature and a mobile phase consisted of 1% aqueous acetic acid containing 0.2% triethylamine and acetonitrile (65:35, v/v; pH 3.8). The flow-rate was kept at 0.5 mL/min and column effluent was monitored with a fluorescence detector at an excitation wavelength of 230 nm and an emission wavelength of 340 nm. Retention times for pronethalol and propranolol were 8.5 min and 10.5 min, respectively. Linear regressions for the standard curves were linear in the range 2-800 ng/mL, giving correlation coefficients above 0.998. The detection limit was 1.34 ng/mL. No analytical interference was observed from endogenous components in rat plasma. This simple and sensitive assay method was feasibly applied to the pharmacokinetic study of propranolol after intravenous administration of 2 mg/kg of propranolol to normal and carbon tetrachloride-induced liver cirrhotic rats.

Adrenergic beta-Antagonists↗

Fine-needle aspiration cytology of pleuropulmonary blastoma: a case report with unusual features.

Pleuropulmonary blastoma (PPB) is a rare and aggressive intrathoracic neoplasm of childhood, typically presenting as a pulmonary and/or pleural-based mass with cystic, solid, or combined features. Histologically, the tumor is well characterized with a mixture of primitive mesenchymal and variably differentiated sarcomatous components, and the cytologic features described in the three previous reports are also compatible to the histologic ones. Now, we present another case of PPB showing unusual features. A 3-yr-old boy presented with a pleural- or chest wall-based tumor. The fine-needle aspiration cytologic smears were highly cellular with poorly differentiated cells showing variable size and shape rather than those of typical blastemal cells. The histologic findings were also distinctive, exhibiting diffuse sheets of poorly differentiated cells without typical blastemal cell component. Diagnosis was confirmed by the support of immunohistochemical and ultrastructural features. The patient underwent a typical aggressive clinical course to death within 8 mo after diagnosis.

Biomarkers, Tumor↗

Prognostic factors of Krukenberg's tumor.

OBJECTIVE: The aim of this study was to determine prognostic factors of metachronous Krukenberg's tumors of gastric origin, thereby helping to establish a therapeutic plan for this rare entity. METHODS: Thirty-four female patients who underwent curative resection of gastric carcinoma from 1987 through 1996 and subsequently developed Krukenberg's tumors were identified. The covariates used for survival analysis were patient age at the time of ovarian relapse (categorized as <50 years or >/=50 years), size of Krukenberg's tumor (categorized as </=5, 5-10, or >10 cm), initial AJCC stage of gastric carcinoma, relapse-free interval (categorized as <1, 1-2, or >/=2 years), extent of recurrent disease, and the presence of gross residual disease after treatment for Krukenberg's tumors. The influence of all covariates on the survival period was assessed using the Cox proportional hazards model. RESULTS: At a median follow-up of 32.3 months (range, 2.2-83.1), the median survival period of 34 study patients was estimated to be 7.7 months (95% confidence interval (CI), 5.8-9.6) after the diagnosis of metachronous Krukenberg's tumor. Cox-model-based survival analysis demonstrated that the absence of residual disease after treatment and limited disease extent were favorable prognostic factors. The median survival period of patients without gross residual disease was longer (10.9 months (95% CI, 4.6-17.1)) than that of patients with gross residual disease (7.5 months (95% CI, 6.6-8.3)) [P = 0.036]. Median survival periods according to the extent of metastasis were 10.9 months (95% CI, 0-23.0) for patients with disease confined to the ovaries, 13.1 months (95% CI, 5.9-20.3) for patients with disease confined to the pelvis, 7.5 months (95% CI, 6.8-8.1) for patients with intraabdominal disease, and 3.6 months (95% CI, 3.4-3.7) for patients with disease spread outside the abdomen and pelvis [P = 0.003]. In contrast, patient age, size of ovarian tumor, initial stage of gastric adenocarcinoma, and relapse-free interval were not prognostic indicators for survival after the development of ovarian metastases. CONCLUSION: The absence of residual disease after treatment and limited disease extent were favorable prognostic factors of metachronous Krukenberg's tumors of stomach origin. Our data suggest that resection might have a role in the management of Krukenberg's tumors of stomach origin if it could render patients free of gross residual disease.

Adult↗

Expression in Escherichia coli of the thermostable inorganic pyrophosphatase from the Aquifex aeolicus and purification and characterization of the recombinant enzyme.

The gene encoding the inorganic pyrophosphatase from a hyperthermophilic bacterium, Aquifex aeolicus (Aae), was amplified by PCR. Then, the gene was overexpressed in Escherichia coli using a pJR-based expression plasmid, pAIPD. The recombinant Aae pyrophosphatase was purified 16.2-fold with a 53.4% yield and a specific activity of 34 U/mg protein by a combination of heating (to denature E. coli proteins) and two steps of DEAE-Sephacel column chromatography (nonabsorbed enzyme at pH 7.3 and absorbed enzyme at pH 8.0). This enzyme has an approximate molecular mass of 105,000 Da and consists of four subunits, each with a molecular mass of 24,500 Da. The enzyme shows the optimal activity in the pH range 7.5-8.0. The enzyme was stable at 80-95 degrees C. A divalent cation was absolutely required for the enzyme activity, Mg(2+) being most effective.

Bacteria↗

Porcine factor V: cDNA cloning, gene mapping, three-dimensional protein modeling of membrane binding sites and comparative anatomy of domains.

Factor V is a plasma protein essential for blood coagulation. This protein is involved in activated protein C resistance, the most common inherited thrombotic disorder known. We utilized the polymerase chain reaction to clone the porcine factor V gene by generating overlapping clones amplified with primers chosen by comparison with known nucleotide sequences. The porcine factor V cDNA contig encodes a predicted 2258-amino acid protein, making it the largest in comparison to the bovine, human, and murine proteins. Porcine factor V has the highest level of homology with bovine factor V, but also has high levels of conservation of important residues with all the species. Radiation hybrid mapping assigned the porcine factor V gene to chromosome 4. Three-dimensional models of factor V were generated and used to analyze membrane-binding sites in terms of conserved, and therefore likely important residues.

Amino Acid Sequence↗

Expression of the seqA gene is negatively modulated by the HU protein in Escherichia coli.

The SeqA protein acts as a regulator of chromosomal replication initiation in Escherichia coli by sequestering hemi-methylated oriC, effectively blocking methylation and therefore preventing rapid re-initiation. The level of SeqA protein is maximal at mid-log phase and decreases when cells enter late-log phase. In hup mutants that lack the HU protein, the maximal seqA expression is also seen at mid-log phase, but seqA expression, as well as SeqA levels and activity, is increased by up to four fold relative to that in the wild type. These results suggest that the HU protein functions as a negative modulator of seqA expression.

Bacterial Outer Membrane Proteins↗

Accumulation of 2,5-dimethoxy-1,4-benzoquinone in suspension cultures of Panax ginseng by a fungal elicitor preparation and a yeast elicitor preparation.

Suspension cultures of Panax ginseng C.A. Meyer (Araliaceae) were treated with either an elicitor preparation from the culture broth of the phytopathogenic hyphomycete Botrytis cinerea or a yeast elicitor preparation, and the accumulation of a new compound, which was not detected in non-elicited cultures, was observed. The accumulated compound was isolated and shown to be 2,5-dimethoxy-1,4-benzoquinone by 1H-NMR, 13C-NMR and electron ionization (EI) mass spectra. While it is well known that this compound shows antibacterial activity against Staphylococcus aureus, its presence in ginseng root has not been reported to date. Levels of the compound in the media increased rapidly, reaching a maximum level of 65.10 +/- 4.96 microg/g fresh weight at approximately 12 h after treatment with the yeast elicitor preparation. The maximal level of the compound in medium from the culture treated with an elicitor preparation from the culture broth of B. cinerea was 46.13 +/- 10.42 microg/g fresh weight after 24 h of incubation.

Benzoquinones↗

Identification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis.

The functional importance of a conserved region in a novel chitosanase from Bacillus sp. CK4 was investigated. Each of the three carboxylic amino acid residues (Glu-50, Glu-62, and Asp-66) was changed to Asp and Gln or Asn and Glu by site-directed mutagenesis, respectively. The Asp-66-->Asn and Asp-66-->Glu mutation remarkably decreased kinetic parameters such as Vmax and kcat to approximately 1/1,000 those of the wild-type enzyme, indicating that the Asp-66 residue was essential for catalysis. The thermostable chitosanase contains three Cys residues at positions 49, 72, and 211. The Cys-49-->Ser/Tyr and Cys-72-->Ser/Tyr mutant enzymes were as stable to thermal inactivation and denaturating agents as the wild-type enzyme. However, the half-life of the Cys-211-->Ser/Tyr mutant enzyme was less than 10 min at 80 degrees C, while that of the wild-type enzyme was about 90 min. Moreover, the residual activity of Cys-211-->Ser/Tyr enzyme was substantially decreased by 8 M urea; and it lost all catalytic activity in 40% ethanol. These results show that the substitution of Cys with any amino acid residues at position 211 seems to affect the conformational stability of the chitosanase.

Amino Acid Sequence↗

Case report: diabetic muscle infarction presenting as knee arthralgia.

Diabetic muscle infarction is a rare complication of diabetes and has characteristic clinical features including acute onset of pain with painful swelling, most commonly in the thigh or calf muscle, which gradually improves to complete resolution. Recently we experienced a case of diabetic muscular infarction presenting as knee joint pain due to involvement of the proximal portion of the leg muscle, which site has not been reported previously. This case shows that diabetic muscle infarction may involve sites other than the thigh and calf areas and should be considered in the differential diagnosis of knee arthralgia.

Aged↗

Dietary alpha-linolenic acid lowers postprandial lipid levels with increase of eicosapentaenoic and docosahexaenoic acid contents in rat hepatic membrane.

This study was designed to examine the effects of dietary n-3 and n-6 polyunsaturated fatty acids (PUFA) on postprandial lipid levels and fatty acid composition of hepatic membranes. Male Sprague-Dawley rats were trained for a 3-h feeding protocol and fed one of five semipurified diets: one fat-free diet or one of four diets supplemented with 10% (by weight) each of corn oil, beef tallow, perilla oil, and fish oil. Two separate experiments were performed, 4-wk long-term and 4-d short-term feeding models, to compare the effects of feeding periods. Postprandial plasma lipid was affected by dietary fats. Triacylglycerol (TG) and total cholesterol levels were decreased in rats fed perilla oil and fish oil diets compared with corn oil and beef tallow diets. Hepatic TG and total cholesterol levels were also reduced by fish oil and perilla oil diets. Fatty acid composition of hepatic microsomal fraction reflected dietary fatty acids and their metabolic conversion. The major fatty acids of rats fed the beef tallow diet were palmitic, stearic, and oleic. Similarly, linoleic acid (LA) and arachidonic acid in the corn oil group, alpha-linolenic acid (ALA) and eicosapentaenoic acid (EPA) in the perilla oil group, and palmitic acid and docosahexaenoic acid (DHA) in the fish oil group were detected in high proportions. Both long- and short-term feeding experiments showed similar results. In addition, microsomal DHA content was negatively correlated with plasma lipid levels. Hepatic lipid levels were also negatively correlated with EPA and DHA contents. These results suggest that n-3 ALA has more of a hypolipidemic effect than n-6 LA and that the hypolipidemic effect of n-3 PUFA may be partly related to the increase of EPA and DHA in hepatic membrane.

Animals↗

The role of the pars recta and pars oblique of cricothyroid muscle in speech production.

To evaluate the functional difference of the pars recta and pars oblique during speech production, the electromyographic activities of these muscles were measured in thyroidectomized patients. The hooked wire electrodes were inserted into the normal side of the bellies of the pars recta and pars oblique bundles. Two kinds of sentences were used to obtain pitch changes, a simple interrogative sentence and a complex sentence with stress contrasts. The pars recta and pars oblique were simultaneously activated for initial lengthening and tensing of vocal folds to produce speech. The pars oblique might be initially more active than the pars recta at the initial task of speech and the pars recta might be more active at the pitch elevation in the interrogative sentence and the stress contrast of the complex sentence. The maximum electromyographic activity range of the pars recta and pars oblique seemed to be nearly equal. These results demonstrated that the patterns of electrical activities of the two bellies are different during speech and the combined activities of the pars recta and pars oblique are important in the adjustment of the vocal fold length during speech.

Adult↗

Molecular identification of Oxalobacter formigenes with the polymerase chain reaction in fresh or frozen fecal samples.

OBJECTIVE: To develop a simple and rapid polymerase chain reaction (PCR) method for detecting Oxalobacter formigenes (which degrades oxalate in the gut) in fecal specimens from healthy volunteers and patients with urolithiasis, and to determine whether O. formigenes can be detected in frozen or fresh fecal samples. MATERIALS AND METHODS: Whole bacterial DNA was isolated directly from fresh and frozen fecal samples obtained from 30 healthy volunteers free from urolithiasis and from fresh fecal samples obtained from 38 patients with urolithiasis. Genus-specific oligonucleotide sequences were designed, corresponding to homologous regions residing in the oxc gene that encodes for oxalyl-coenzyme A decarboxylase. A PCR-based assay was used on both fresh and frozen fecal samples, and the nucleotide sequences analysed to confirm oxc. RESULTS: A PCR product of 416 bp encoding the oxc gene was detected in 23 (77%) of 30 healthy volunteers free from urolithiasis and in 14 (37%) of 38 patients with urolithiasis. In healthy volunteers, the results of PCR for the fresh and the frozen samples were identical in each subject. The nucleotide sequence analysis showed that the sequence of the amplified product was compatible with that of oxc. CONCLUSION: O. formigenes can be identified easily and efficiently using this PCR-based detection system. The colonization rate of O. formigenes in patients with urolithiasis was significantly lower than that in healthy volunteers known to be free from urolithiasis. Furthermore, as the PCR-based assay results in the frozen fecal samples were identical to those from fresh samples in each subject, immediate processing of fecal samples may not be necessary to detect O. formigenes in the clinical setting.

Adult↗

Cyclin E overexpression as an independent risk factor of visceral relapse in breast cancer.

AIM: Prognostic value of the cyclin E overexpression in breast cancer has not been clearly established, especially in relation to the pattern of recurrence. We investigated the implication of cyclin E overexpression for the pattern of recurrence in Korean breast cancer patients. METHODS: Using immunohistochemical methods, we retrospectively examined the cyclin E expression level in breast cancer specimens from 128 women who underwent curative breast surgery, and correlated the levels of expression with the pattern of relapse in patients. RESULTS: Cox model-based multivariate analysis indicated that distant relapse could be predicted by the number of positive axillary lymph nodes, high cyclin E expression, and the younger age (<35 years) of the patient. We tested further the association of cyclin E overexpression with the specific types of recurrence; multivariate analyses indicated that adjusted relative risks of bone and visceral relapse as the first events among high cyclin E group were 2.46 (95% confidence interval (CI), 0.86-7.02) (P=0.092), and 3.98 (95% CI, 1.23-12.94) (P=0.022), respectively. On the other hand, cyclin E overexpression was not associated with the risk of locoregional relapse. CONCLUSION: Our data suggest that cyclin E overexpression in primary breast carcinoma tissue could independently predict the risk of distant relapse, especially of visceral relapse, as the first failure after curative breast surgery.

Adult↗

Arthroscopic anterior cruciate ligament reconstruction with quadriceps tendon composite autograft.

At present, no single graft option clearly outperforms another. Autografts (patellar tendon, hamstring) and allografts (Achilles tendon, patellar tendon) are the grafts most often used. However, each grafts has advantages and disadvantages. Quadriceps tendon graft for anterior cruciate ligament reconstruction is not new, but an alternative composite graft is introduced here that consists of quadriceps tendon-patellar bone and bone obtained from a coring reamer used to create the tibial tunnel. This composite graft retains reduced morbidity while allowing the secure bone-to-bone fixation associated with bone-patellar tendon-bone graft.

Anterior Cruciate Ligament↗