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Biomedical subjects

J H Shin

Publications and source records attributed to J H Shin.

At least 91 records · Page 5Linked to original sources
PubMed1999

4-Aminopyridine (4-AP) augments Ca(2+)-dependent action potential and changes oscillatory firing patterns in rat cerebellar Purkinje cells.

Intracellular recordings in cerebellar slice preparation showed that applications of 4-AP altered the pattern of oscillatory firing activity in Purkinje cells (PCs), especially yielding pronounced changes in action potential shape. 4-AP increased the amplitude and duration of action potential significantly and decreased the spike frequency. After 4-AP application, the duration of bursting was prolonged and the duration of after-burst hyperpolarization was progressively shortened. In all PCs tested, the rhythmicity of oscillatory firing activity was abolished completely at the steady state. These results suggest that 4-AP-sensitive currents determine the shape and frequency of individual Ca(2+)-dependent action potentials as well as maintaining oscillatory firing activity in PCs.

4-Aminopyridine↗
PubMed1999

Changes in the evolution of the antigenic profiles and morphology during coccoid conversion of Helicobacter pylori.

OBJECTIVES: The significance of the coccoid forms of H. pylori is still controversial and the questions of whether these forms are viable and infective or degenerative are still open. We induced conversion from rod to coccoid forms and studied morphological changes and antigenic evolutions during this conversion and, thereby, elucidated the viability of coccoid forms. METHODS: The H. pylori strain (C001) used for Western blotting was isolated from the patient with gastric cancer. The antigenic evolution during coccoid conversion of H. pylori was studied by Western blotting, using different sera from thirty patients known to be culture positive. These sera were used to reveal the total antigens of the strain cultured for 2 days (100% rod) and 15 days (> 99% coccoid). After SDS-PAGE, with 10% separating gel of total antigens (rod and coccoid), transblotting (Trans-Blot electrophoretic cell, Bio-Rad) was taken onto a nitrocellulose membrane (Bio-Rad). Then, the blots, with human sera diluted at 1/100, were developed with color reaction by goat serum anti-human IgG with alkaline phosphatase and BCIP. RESULTS: The antigenic profiles were not changed in 46.7% (14/30 cases) and were changed in 53.3% (16/30 cases) during coccoid conversion. Antigenic fractions changed during coccoid conversion were protein band at 120 kDa and band at 35 kDa, and were not detected in coccus forms. The rest of the profiles were identical between rod and coccoid forms. The protein which disappeared include CagA (120 kDa) and porin, or adhesin (35 kDa). The morphological changes during coccoid conversion were U shaped at day 7, doughnut shaped at day 9 and full coccoid at day 15. CONCLUSIONS: The results showed that coccoid forms of H. pylori retain cellular structures similar to rod form, and some of the antigens (CagA and porin) disappeared during coccoid conversion. Therefore, coccoid form might be viable and represent one of the stages of H. pylori biological cycle.

Adaptation, Physiological↗
PubMed1999

Molecular typing of vibrio vulnificus isolated from clinical specimens by pulsed-field gel electrophoresis and random amplified polymorphic DNA analysis.

Primary V. vulnificus septicemia has been continuously reported in Korea. We analyzed the molecular diversity of V. vulnificus strains isolated from clinical specimens using pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) analysis. We analyzed a total of 23 V. vulnificus strains isolated from 22 patients between 1992 and 1997. RAPD analysis was performed using five random primers, and we obtained chromosomal DNA restriction patterns with NotI and SfiI by PFGE. Two isolates from one patient disclosed similarity value 1.0 by RAPD and had an indistinguishable pattern when analyzed with PFGE, which indicated that they were the same strains. The remaining 22 isolates from 22 patients could be separated into 5 different molecular types in RAPD analysis. The range of similarity values among the isolates was wide (0.29-0.81). Of 22 strains, four strains could not be typed in repeated trials by PFGE with NotI and SfiI. The PFGE patterns of 18 strains showed a remarkable polymorphism consisting 12-19 fragments (20-870 kb). These results show that V. vulnificus strains isolated from Korea are genetically diversified for the most part.

Adult↗
PubMed1998

Analysis of the gene encoding cyclomaltodextrinase from alkalophilic Bacillus sp. I-5 and characterization of enzymatic properties.

The gene encoding cyclomaltodextrinase (CDase) was cloned from alkalophilic Bacillus sp. I-5. The nucleotide sequence of the gene was determined and the physicochemical properties of the enzyme were investigated. The gene had an open reading frame of 559 amino acids with a predicted molecular weight of 64,884. The enzyme was purified to near homogeneity from Escherichia coli cells carrying a recombinant plasmid that contained the CDase gene. The enzyme hydrolyzed cyclomaltoheptaose (beta-CD) 13 times better than starch and 33 times better than pullulan, and it had transglycosylation activity. The enzyme also hydrolyzed acarbose, a pseudotetrasaccharide inhibitor of glucosidases. The enzyme was stabilized by Ca2+ and the activity was increased more than twofold in the presence of 5 mM EDTA. The optimum temperature of the enzyme was elevated from 40 to 50 degrees C by Ca2+ ion and the thermal activity was maintained more than 80% at 60 degrees C in the presence of Ca2+. Comparison of known amino acid sequences of several amylolytic enzymes with cyclomaltodextrinase activity, site-directed mutagenesis of the enzyme, and substrate specificity of the enzyme imply that the region between the third and the fourth conserved regions of the enzyme may play an important role in binding and degradation of cyclomaltodextrin.

Acarbose↗
PubMed1998

Direct identification of Vibrio vulnificus in clinical specimens by nested PCR.

This study was performed to establish optimal nested PCR conditions and a high-yield DNA extraction method for the direct identification of Vibrio vulnificus in clinical specimens. We designed two sets of primers targeting the V. vulnificus hemolysin/cytolysin gene. The target of the first primer set (P1-P2; sense, 5'-GAC-TAT-CGC-ATC-AAC-AAC-CG-3', and antisense, 5'-AGG-TAG-CGA-GTA-TTA-CTG-CC-3', respectively) is a 704-bp DNA fragment. The second set (P3-P4; sense, 5'-GCT-ATT-TCA-CCG-CCG-CTC-AC-3', and antisense, 5'-CCG-CAG-AGC-CGT-AAA-CCG-AA-3', respectively) amplifies an internal 222-bp DNA fragment. We developed a direct DNA extraction method that involved boiling the specimen pellet in a 1 mM EDTA-0.5% Triton X-100 solution. The new DNA extraction method was more sensitive and reproducible than other conventional methods. The DNA extraction method guaranteed sensitivity as well, even when V. vulnificus cells were mixed with other bacteria such as Escherichia coli or Staphylococcus aureus. The nested PCR method could detect as little as 1 fg of chromosomal DNA and single CFU of V. vulnificus. We applied the nested PCR protocol to a total of 39 serum specimens and bulla aspirates from septicemic patients. Seventeen (94.4%) of the 18 V. vulnificus culture-positive specimens were positive by the nested PCR. Eight (42.1%) of the 19 culture-negative samples gave positive nested PCR results.

Base Sequence↗
PubMed1998

Fatal Hormonema dematioides peritonitis in a patient on continuous ambulatory peritoneal dialysis: criteria for organism identification and review of other known fungal etiologic agents.

We report a fatal case a fungal peritonitis caused by the yeast-like dematiaceous mould Hormonema dematioides in a 45-year-old woman. The woman had a 13-year history of insulin-dependent diabetes mellitus and had been on continuous ambulatory peritoneal dialysis for chronic renal failure. H. dematioides was repeatedly isolated from the dialysate culture specimens collected on days 3, 9, 16, and 20 of her hospital stay. Preliminary culture reports on day 7 of the growth of a yeast-like fungus, a probable Candida species, prompted the administration of fluconazole (FLU). Intraperitoneal and intravenous FLU failed to eliminate the mould, and the patient expired on day 21 of her hospital stay. We use this case to present what appears to be the first report of fungal peritonitis due to H. dematioides, to provide laboratorians with criteria for differentiating this organism from the similar mould Aureobasidium pullulans and from various yeast genera, and to provide a review of known fungal taxa inciting peritonitis.

Ascitic Fluid↗
PubMed1998

Effects of t-butyl hydrogen peroxide on single SR calcium release channels.

Using lipid bilayer reconstitution technique, we investigated the oxidation effect of t-butyl hydrogen peroxide (tBHP) on the single channel activity of the sarcoplasmic reticulum (SR) calcium release channels isolated from canine latissimus dorsi muscles. When 0.7% tBHP was added in the cytosolic side, the channel activity became suppressed (n = 7), and it was recovered by changing the solution to the control solution. The suppression was due to the change in the gating mode of the channel: before tBHP the channel opened to four sub-conductance levels, but it opened to only one level after tBHP. These effects by tBHP were different from the previous finding using hydrogen peroxide (H2O2), which may be explained by different oxidation patterns between the two oxidants.

Animals↗
PubMed1998

Korean Nationwide Surveillance of Antimicrobial Resistance of bacteria in 1997.

Antimicrobial-resistant bacteria are known to be prevalent in tertiary-care hospitals in Korea. Twenty hospitals participated to this surveillance to determine the nationwide prevalence of resistance bacteria in 1997. Seven per cent and 26% of Escherichia coli and Klebsiella pneumoniae were resistant to 3rd-generation cephalosporin. Increased resistance rates, 19% of Acinetobacter baumannii to ampicillin/sulbactam, and 17% of Pseudomonas aeruginoa to imipenem, were noted. The resistance rate to fluoroquinolone rose to 24% in E. coli, 56% in A. baumannii and 42% in P. aeruginosa. Mean resistance rates were similar in all hospital groups: about 17% of P. aeruginosa to imipenem, 50% of Haemophilus influenzae to ampicillin, 70% of Staphylococcus aureus to methicillin, and 70% of pneumococci to penicillin. In conclusion, nosocomial pathogens and problem resistant organisms are prevalent in smaller hospitals too, indicating nosocomial spread is a significant cause of the increasing prevalence of resistant bacteria in Korea.

Bacterial Physiological Phenomena↗
PubMed1998

A case of arteriovenous type cardiac hemangioma.

Cardiac hemangiomas are rare primary tumors of the heart and constitute only 2.8% of primary cardiac tumors. They are classified into capillary, cavernous, epitheloid and arteriovenous type and the last one is the most uncommon type. We experienced a case of cardiac hemangioma which was diagnosed as arteriovenous type for the first time in Korea in the literature. The patient was a 54-year-old woman who presented with palpitation and anterior chest pain. The diagnosis was based upon coronary angiography which showed two tumor blushings located in the interatrial and interventricular septum with venous drainage to the coronary sinus and right atrium. Associated atrial fibrillation with rapid ventricular response was controlled with digitalis.

Atrial Fibrillation↗
PubMed1997

Nitric oxide directly activates calcium-activated potassium channels from rat brain reconstituted into planar lipid bilayer.

Using the planar lipid bilayer technique, we tested whether NO directly activates calcium-activated potassium (Maxi-K) channels isolated from rat brain. We used streptozotocin (STZ) as NO donor, and the NO release was controlled with light. In the presence of 100-800 microM STZ, the Maxi-K channel activity increased up to 3-fold within several tens of seconds after the light was on, and reversed to the control level several minutes after shutting off the light. Similar activation was observed with other NO donors such as S-nitroso-N-acetylpenicillamine and sodium nitroprusside. The degree of activity increase was dependent upon the initial open probability (P[init]). When the P(init) was lower, the activity increase was greater. These results demonstrate that NO can directly affect the Maxi-K channel activity, and suggest that the Maxi-K channel might be one of the physiological targets of NO in brain.

Action Potentials↗
PubMed1997

Parathyroid hormone-related protein enhances insulin-like growth factor-I expression by fetal rat dermal fibroblasts.

Interactions between cells of differing embryonic origins comprise a common theme during tissue development and repair. Often, communication between them can be mediated by soluble growth mediators and in some cases is restricted in focus. That is, some cells respond to, but do not produce, mediators expressed by other cells within the tissue. Because keratinocytes respond to but do not express insulin-like growth factor I (IGF-I), another skin cell population, the dermal fibroblast, may supply this factor. However, keratinocytes express, but do not respond to parathyroid hormone related protein (PTHrp), which increases cAMP production by dermal fibroblasts. Based on earlier results where inducers of cAMP increase local IGF-I expression in skeletal tissue, we postulated that PTHrp might induce local IGF-I by dermal fibroblasts and provide a source of this factor for keratinocyte activity. Our studies reveal that IGF-I mRNA and protein levels increase in response to PTHrp in vitro, and that this effect is replicated by inducers of cAMP, but not by activators of protein kinase C. Consequently, these factors appear to comprise a paracrine loop within the skin, permitting focused but restricted IGF-I expression to support skin growth, remodeling, or repair.

Animals↗
PubMed1997

Eugenol protects neuronal cells from excitotoxic and oxidative injury in primary cortical cultures.

We examined the neuroprotective efficacy of eugenol against N-methyl-D-aspartate (NMDA)-, oxygen-glucose deprivation-, and xanthine/xanthine oxidase-induced neurotoxicity in primary murine cortical cultures. Eugenol (100-300 microM) attenuated NMDA (300 microM)-induced acute neurotoxicity by 20-60%. At the same concentration range, eugenol also inhibited NMDA (300 microM)-induced elevation in neuronal 45Ca2+ uptake by 10-30%. In the oxygen-glucose deprivation (50 min) neurotoxicity, eugenol (100-300 microM) prevented acute neuronal swelling and reduced neuronal death by 45-60% in a concentration-dependent fashion. Oxidative neuronal injury induced by xanthine/xanthine oxidase was also significantly reduced (75-90%) by eugenol (100- 300 microM) addition. These results suggest that eugenol may play a protective role against ischemic injury by modulating both NMDA receptor and superoxide radical.

Animals↗
PubMed1997

Rapid identification of Candida species in blood cultures by a clinically useful PCR method.

Widespread use of fluconazole for the prophylaxis and treatment of candidiasis has led to a reduction in the number of cases of candidemia caused by Candida albicans but has also resulted in the emergence of candidemias caused by innately fluconazole-resistant, non-C. albicans Candida species. Given the fulminant and rapidly fatal outcome of acute disseminated candidiasis, rapid identification of newly emerging Candida species in blood culture is critical for the implementation of appropriately targeted antifungal drug therapy. Therefore, we used a PCR-based assay to rapidly identify Candida species from positive blood culture bottles. This assay used fungus-specific, universal primers for DNA amplification and species-specific probes to identify C. albicans, C. krusei, C. parapsilosis, C. tropicalis, or C. glabrata amplicons. It also used a simpler and more rapid (1.5-h) sample preparation technique than those described previously and used detergent, heat, and mechanical breakage to recover Candida species DNA from blood cultures. A simple and rapid (3.5-h) enzyme immunosorbent assay (EIA)-based format was then used for amplicon detection. One hundred fifty blood culture bottles, including 73 positive blood culture bottle sets (aerobic and anaerobic) from 31 patients with candidemia, were tested. The combined PCR and EIA methods (PCR-EIA) correctly identified all Candida species in 73 blood culture bottle sets, including bottles containing bacteria coisolated with yeasts and 3 cultures of samples from patients with mixed candidemias originally identified as single-species infections by routine phenotypic identification methods. Species identification time was reduced from a mean of 3.5 days by routine phenotypic methods to 7 h by the PCR-EIA method. No false-positive results were obtained for patients with bacteremias (n = 18), artificially produced non-Candida fungemias (n = 3), or bottles with no growth (n = 20). Analytical sensitivity was 1 cell per 2-microl sample. This method is simpler and more rapid than previously described molecular identification methods, can identify all five of the most medically important Candida species, and has the potential to be automated for use in the clinical microbiology laboratory.

Candida↗
PubMed1997

Molecular epidemiologic analysis of Staphylococcus aureus isolated from clinical specimens.

Nosocomial infections caused by Staphylococcus aureus are clinically serious and control of such infections requires strain typing to identify the source of contamination. Recently, pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) assay have been introduced and have provided a high level of strain discrimination of S. aureus isolated from clinical specimens. This study was performed to classify 82 strains of S. aureus isolated from 4 hospitals in the Kwangju-Chonnam area by PFGE and RAPD assay. Methicillin-resistant S. aureus (MRSA) was identified by disk diffusion method using the oxacillin disk and polymerase chain reaction of mecA gene was done in 69 strains. Eight-three strains including S. aureus ATCC 25923 were classified into 10 groups by RAPD assay, and into 8 groups by PFGE. Classified groups were not related to area or hospital. Classification was not characteristic between MRSA and methicillin-susceptible strains. Nosocomial infections due to outbreak were suggested because some strains disclosed identical band patterns by PFGE. These results indicate that medical personnels and instruments are routes of nosocomial infections caused by MRSA. PFGE and RAPD assay are powerful tools for the epidemiological study of S. aureus, but PFGE is more effective than RAPD assay. RAPD assay needs optimal combination of primers.

Bacterial Proteins↗
PubMed1997

Aspergillus nidulans infection in a patient with chronic granulomatous disease.

Aspergillus nidulans is one of the several species of Aspergillus with low pathogenicity. The significant infections of A. nidulans in human have rarely been reported, almost exclusively in patients with chronic granulomatous disease (CGD). CGD is a primary immunodeficiency disease which results from the absence of the NADPH oxidase in the phagocytic cells, leading to recurrent pyogenic infection and granuloma and abscess formation. Here we report a fatal case A. nidulans infection in a six-year-old boy with chronic granulomatous disease. A. nidulans was isolated from the culture of a paraspinal abscess and Aspergillus was detected in the surgical tissue by in situ hybridization. The patient succumbed despite prolonged treatment with high-dose amphotericin B, itraconazole and interferon-alpha. To our knowledge, this is the first report of A. nidulans infection in Korea.

Aspergillosis↗