BACKGROUND: Nineteen strains of Shigella sonnei isolated from the patients were examined regarding their biochemical characterization, serotype, and antibiotics resistance, and then analyzed for plasmid DNA profile. METHODS: Strains were tested for possession of set1A, set1B, sen, ipaH, ial, stx and invE genes using the polymerase chain reaction (PCR) method and were analyzed using the pulsed-field gel electrophoresis (PFGE) pattern against 7 outbreak isolates (10 strains). RESULTS: These strains had the typical biochemical characterization of S. sonnei with positive ornithine decarboxylase and -galactosidase activity, but were negative in mannitol fermentation. Serotype were identified as the I phase in 13 strains (68.0%) and the II phase in 6 strains (32.0%). All strains were resistant to erythromycin, vancomycin, tetracycline, and penicillin. The antibiogram type showed 4 groups from I to IV. The strains showed 8 types of plasmid profiles and were designated as P1 to P8. By the PCR, the ipaH gene and the set1B gene were detected from all of the 16 strains. The invE was detected from 9 strains (56.3%), and the sen gene was detected from 5 strains. All strains were negative for the Stx and the set1A gene. High-molecular-weight genomic DNA was prepared from 7 outbreak isolates (10 strains) and digested with the restriction endonuclease XbaI. Restriction fragment patterns of chromosomal DNA were demonstrated by PFGE. XbaI produced about 23 fragments in all strains with the their size ranged from 40 to 680 kb. Ten strains could be differentiated to 3 patterns by chromosomal DNA fingerprint. CONCLUSIONS: All of the Shigella sonnei strains that were isolated from Busan Province showed similar chromosomal DNA fragment patterns, while the Japanese differed in chromosomal DNA fingerprint pattern. PFGE is useful for the epidemiological study of Shigella sonnei associated endemic diarrhea.
Anti-Bacterial Agents ; Asian Continental Ancestry Group ; Busan ; Diarrhea ; DNA ; DNA Fingerprinting ; DNA Restriction Enzymes ; Electrophoresis, Gel, Pulsed-Field ; Erythromycin ; Fermentation ; Genotype ; Humans ; Mannitol ; Microbial Sensitivity Tests ; Ornithine Decarboxylase ; Penicillins ; Plasmids ; Polymerase Chain Reaction ; Shigella sonnei* ; Shigella* ; Tetracycline ; Vancomycin ; Virulence Factors* ; Virulence*

Staphylococcus aureus continues to be the main cause of surgical site infections. Recently, methicillin-resistant S. aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes the problem of neonatal nosocomial infection. Antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions. Therefore it is necessary to develop a new method based on a molecular biological technique in order to overcome these problems. We report the development of a multiplex PCR protocol for the diagnosis of staphylococcal infection. The protocol was designed to i) detect any staphylococcal species to the exclusion of other bacterial pathogens (based on primers corresponding to staphylococcal-specific regions of the 16S rRNA genes), ii) provide an indication of the likelihood that the Staphylococci present in the specimen are resistant to oxacillin (based on the amplication of the mecA gene, encoding penicillin-binding protein 2'(PBP-2'), which is known to confer resistance to the bacteriostatic action of methicillin). In this study, 67 S. aureus strains were isolated from the neonatal intensive care unit and general neonatal nursery at Pusan National University Hospital, Busan, Korea, between January and July 2003. Methicillin resistance was tested by the oxacillin disk diffusion method and the MIC method. We performed the multiplex PCR to amplify the mecA gene, encoding PBP-2'. We tested it by multiplex PCR and compared the results with the antimicrobial susceptibilities. Different results were obtained from 2 MRSA (4.65%), suggesting that the PCR method should be performed at the same time for a more accurate clinical test of MRSA
Anti-Bacterial Agents ; Busan ; Cross Infection ; Diagnosis ; Diffusion ; Infant, Newborn ; Intensive Care, Neonatal ; Korea ; Methicillin Resistance* ; Methicillin-Resistant Staphylococcus aureus* ; Multiplex Polymerase Chain Reaction* ; Nurseries ; Oxacillin ; Penicillin-Binding Proteins ; Polymerase Chain Reaction ; Staphylococcal Infections ; Staphylococcus aureus ; Thiram

Agenesis of trachea is a rare congenital anomaly. The main signs are respiratory distress, cyanosis, inability to vocalise and impossible tracheal intubation. There is no long-term surgical solution at present, therefore the condition is ultimately fatal. We report a case of tracheal agenesis. We anesthetized a 2.25 kg neonate for endoscopic examination, who was intubated successfully. Beyond 2.5 cm from the vocal cord, there was no trachea and we can only see esophageal lumen with fistula opening. After the endoscopic examination, exploratory thoracotomy was done, but surgical correction was impossible. After the surgery, she died from progressive ventilatory failure. Autopsy revealed a Floyd's type II tracheal agenesis with tracheo-esophageal fistula.
Autopsy ; Cyanosis ; Fistula ; Humans ; Infant, Newborn ; Intubation ; Thoracotomy ; Trachea ; Vocal Cords

PURPOSE: Nosocomial infection with Staphylococcus aureus, especially methicillin resistant S. aureus, has become a serious concern in the neonatal intensive care unit. The aim of this study is to investigate the virulence factors, and the relationship between the antibiotic resistance and the associated genes of Staphylococcus aureus isolated from nasal cavity of neonates. METHODS: Fifty one isolates of S. aureus were obtained from nasal swab taken in 28 neonates in the NICU and nursery of Pusan National University Hospital between February and May, 2001. They were tested in regard to antibiotic susceptibility, coagulase test and typing, plasmid DNA profile, as well as reactivity to enterotoxin A-E(sea, seb, sec, sed, see) genes and toxic shock syndrome toxin-1(tst) gene by polymerase chain reaction(PCR). Associated genes such as mecA, mecR1, mecI, and femA were also determined by PCR. The origin of MRSA strains was assessed using DNA fingerprinting by arbitrarily-primed polymerase chain reaction(AP-PCR). RESULTS: Twenty three(45.1%) and six(11.8%) isolates were resistant to oxacillin and vancomycin respectively. Multidrug resistance to three or more of the antibiotics tested was observed in 51.0% of the isolates. Forty two isolates were coagulase positive and twenty two isolates had mecA gene. Sixteen isolates had both mecA and femA genes and had type I-III plasmids. 64.7% of isolates carried sec gene, and 80.4% carried tst gene. DNA fingerprinting by AP-PCR for 12 MRSA strains showed 10 distinct patterns, suggesting different origins. CONCLUSION: We confirmed that the prevalence of nasal carriage of S. aureus and the incidence of antimicrobial-resistant S. aureus, especially vancomycin resistance, is very high in neonates who were admitted in NICU and nursery. It is possible that these pathogens are responsible for serious nosocomial infections in neonates. The need for improved surveillance and continuous control of pathogens is emphasized.
Infant, Newborn ; Humans ; Incidence

An open clinical trial was performed to test the efficacy and side effects of Pravastatin(Mevlotin(R)), HMG-CoA reductase inhibitor, administering 5mg twice daily for 12weeks in 30 patients of hypercholesterolemia in out patient clinics, Pusan National University Hospital. The total cholesterol, triglyceride and HDL-cholesterol were measured with enzyme methods and LDL-cholesterol was calculated indirectly by Friedewald formula. The result obtained were as follows: 1) The degree of change at the end points compared with baseline pretreatment levels were 26.1% fall in serum total cholesterol.36.6% fall in LDL-cholesterol, 20.8% fall in triglyceride and 14, 6% rise in HDL-cholesterol. And the rate of improvement more than moderate degree were 90.0% in total cholesterol(the fall of 10% or more), 53.3% in triglyceride (the fall 20% or more) and 33.3% in HDL-cholesterol(the rise of 7mg% or more). 2) The total cholesterol/HDL-cholesterol and LDL-cholesterol/HDL cholesterol ratios were decreased significantly from 6, 4+/-0.7 to 4.2+/-0.5(34.4%) and from 4.5+/-0.7 to 2.5 +/-0.4(44.4%) respectively. 3) The greatest fall in serum total cholesterol and LDL-cholesterol were observed in 2 weeks after administrating drug and thereafter fell gradually and maintained until 12 weeks of endpoint, but HDL-cholesterol showed significant rise from the 4 weeks of administration. On the other hand triglyceride showed remarkable fall in the measured values from the 4 weeks but statistical significance was observed only in 10 and 12 weeks after administration owing to wide individual variation of values. 4) There observed the tendency that the higher the initial pretreatment levels the greater the degree of fall in total cholesterol and triglyceride. 5) Neither side effects nor abnormal laboratory findings were shown during the period of observation. The results suggest that Pravastatin will be a useful and safe drug in the treatment of hyperlipidemia.
Busan ; Cholesterol ; Hand ; Humans ; Hypercholesterolemia* ; Hyperlipidemias ; Oxidoreductases ; Pravastatin ; Triglycerides

Purpose: To evaluate the accuracy of intraocular pressure (IOP) measurements obtained by a rebound and non-contact tonometer in eyes with a therapeutic contact lens (CL) after vitrectomy. Methods: In 60 eyes of 60 patients who underwent vitrectomy for vitreoretinal disease, IOP was measured using a rebound tonometer (iCare ic200®; IOPRT) and non-contact computerized air puff tonometer (CT-80, IOPNCT), before and after wearing a CL (Purevision2®, +0.0 diopter). The mean IOP of three consecutive measurements were analyzed, and a comparative analysis with IOP measured by a Goldman applanation tonometer (IOPGAT) was performed. Results: The mean IOPRT without and with the CL was 12.55 ± 5.43 and 13.12 ± 5.13 mmHg, respectively, showing a statistically significant difference (p = 0.02) and strong positive correlation (r = 0.90, p < 0.001). The mean IOPNCT with and without the CL was 12.18 ± 3.24 and 12.17 ± 3.14 mmHg, showing no statistically significant difference (p = 0.17). The consistency with IOPGAT (12.57 ± 5.22 mmHg) was highest in IOPRT without the CL, followed by IOPRT with the CL, IOPNCT without the CL, and IOPNCT with the CL (intraclass correlation coefficients = 0.955, 0.945, 0.856, and 0.850, respectively). In addition, the rebound tonometer successfully measured IOP, regardless of whether the CL was worn; however, the non-contact tonometer failed to measure IOP in seven eyes without the CL and nine with the CL. No difference was observed according to intraocular tamponade type. Conclusions A rebound tonometer can be used as an alternative IOL measuring method in eyes for which it is difficult to use a Goldman applanation tonometer due to the postoperative presence of a therapeutic CL.

A total of 136 strains of Escherichia coli, isolated from rectal swabs of neonates at the neonatal intensive care unit of Pusan National University Hospital during the period from February to April of 2001 and March to April of 2002 were serotyped. The presence of eaeA, aggA, bfpA, astA, LT, and ST genes was test by PCR. Four enteroaggregative E. coli (EAggEC) strains were isolated in 2001 and eight in 2002. In 2001, three strains of enterotoxigenic E. coli (ETEC) were isolated and these strains were tested for antimicrobial susceptibility. ETEC isolates were detected by a reverse passive latex agglutination (RPLA) test for the production of heat-labile enterotoxin. The strains were analyzed by using plasmid profiling, random amplified polymorphic DNA (RAPD), and pulsed field gel electrophoresis (PFGE) methods. The O serotypes could be assigned to 29 isolates (21.3%): O166, 6; O167, 5; O86a, O6 and O127a, 4 each; O8, 2; and O28ac, O44, O158, O20, 1 each. There were 107 untypable isolates. EAggEC isolates were typed as O86a in 4, O127a in 4, and untypable in 4 strains. Three isolates of ETEC were typed as O6. The PCR detected aggA gene in 12 strains (8.8%), but bfpA, eaeA, EAST-1, and ST genes were not detected. LT gene was detected in 3 strains (2.2%) by PCR and DNA probe hybridization, LT production was confirmed in those strains by a latex bead aggregation method. Out of the 15 EAggEC and ETEC strains, eleven (80.0%) were multi-drug resistant to more than 3 antibiotics. Thirteen groups were classed by antibiogram and most strains were susceptible to gentamicin, kanamycin, and cefoperazone, but resistant to ampicillin (100%), cephalothin (66.7%), cefuroxime (46.6%), and tetracycline (46.7%). All isolates possessed a 60 kbp plasmid and 15 strains possessed smaller plasmids. Twelve EAggEC and 3 ETEC strains were differentiated into 5 and 3 groups by the plasmid profiling. RAPD and PFGE grouped the EAggEC isolates into 7 and 6 groups, respectively. The RAPD and PFGE patterns matched in 11 isolates (91.7%) among the 12 EAggEC strains, but the plasmid profile analysis and antibiogram showed no correlation. Three ETEC strains showed different plasmid profiles, and RAPD and PFGE patterns.
Agglutination ; Ampicillin ; Anti-Bacterial Agents ; Busan ; Cefoperazone ; Cefuroxime ; Cephalothin ; DNA ; Electrophoresis, Gel, Pulsed-Field ; Enteropathogenic Escherichia coli* ; Enterotoxigenic Escherichia coli ; Enterotoxins ; Epidemiologic Studies* ; Escherichia coli ; Gentamicins ; Humans ; Infant, Newborn* ; Intensive Care, Neonatal ; Kanamycin ; Latex ; Microbial Sensitivity Tests ; Microspheres ; Plasmids ; Polymerase Chain Reaction ; Tetracycline ; Virulence*

Attaching and effacing Escherichia coli (AEEC) cause enteric infections in humans and animals. Attaching indicates the intimate attachment of bacteria to the enterocyte, and effacing relates to the localized effacement of brush border microvilli. Enteropathogenic (EPEC) and enterohemorrhagic Escherichia coli (EHEC) infections are characterized by the formation of attaching and effacing (AE) lesion on the intestinal epithelial cells. Therefore, they are often grouped together as AEEC. Development of multiplex PCR allowed us to type five of the most important genes implicated in the formation of the AE lesion. A total of 60 AEEC strains isolated from diarrheal patients were investigated by multiplex PCR for the presence of the insertion site of locus of enterocyte effacement (LEE) and LEE-related (eae, tir, espA, espB, and espD) genes. Associating the results of LEE genes typing in the AEEC strains, three different pathotypes are determined: eae(gamma)-tir(gamma)-espA(gamma)-espB(gamma)-espD(gamma) (O157:H7), eae(beta)-tir(beta)-espA(beta)-espB(beta)-espD(beta) (O26:H11), and eae(alpha)-tir(alpha)-espA(alpha)-espB(alpha)-espD(alpha) (O55:H6). These results indicate that AEEC are a heterogenous groups of organisms.
Animals ; Bacteria ; Enterocytes* ; Enterohemorrhagic Escherichia coli* ; Enteropathogenic Escherichia coli* ; Epithelial Cells ; Escherichia coli ; Humans ; Microvilli ; Multiplex Polymerase Chain Reaction

BACKGROUND: Most of the shigellosis outbreaks in Korea have been caused by Shigella sonnei since late 1990's. We analyzed 36 strains of S. sonnei isolated in South Korea from 1998 to 2001 by molecular epidemiologic tools to understand genetic relationship of the outbreaks. METHODS: The 36 strains of S. sonnei were tested for the presence of virulence genes (ial, ipaH, stx, set1A, set1B and sen) using polymerase chain reaction (PCR) method and for the production of Shiga-toxin using latex agglutination test. Seventeen representative strains were selected and their genetic relevance was analyzed by plasmid profile and pulsed-field gel electrophoresis(PFGE). RESULTS: By PCR, ipaH gene was detected in all 36 strains, set1B gene in 15 strains (41.7%), and sen gene in 16 strains (44.4%); all strains were negative for set1A gene. Although stx gene was positive in four strains by PCR method, the toxin was negative by latex agglutination test. The strains were differentiated into 11 groups by plasmid profile and 1 type with 3 subtypes (A-1, A-2and A-3) by PFGE. CONCLUSION: There was a wide range of diverse virulence genes present in the outbreak strains of S. sonnei. PFGE analysis indicated that all the strains tested were related with each other despite minor genotypic and phenotypic differences. A genetically identical clone of S. sonnei was estimated to be the cause of the outbreaks.
Clone Cells ; Disease Outbreaks ; Dysentery, Bacillary ; Genotype ; Korea* ; Latex Fixation Tests ; Molecular Epidemiology* ; Plasmids ; Polymerase Chain Reaction ; Shigella sonnei* ; Shigella* ; Virulence Factors* ; Virulence*

An open clinical trial was conducted to evaluate the efficacy and tolerability of isradipine in 30 cases (male 16, female 14 cases, average age 52.6+/-7.94) of mild to moderate essential hypertension using 1.25-2.5mg twice a day for 8 weeks of active treatment. Blood pressure was significantly reduced from 168.5+/-14.33/108.3+/-6.37mmHg, 163.7+/-9.74/105.5+/-7.1mmHg to 141.0+/-13.69/92.0+/-9.27mmHg, 138.8+/-13.46/92.3+/-11.16mmHg in sitting and standing position respectively. The extent of reduction was 27.5/16.3mmHg in sitting position and 29.9/13.2mmHg in standing position. This comprised the mean response rate in terms of reduction of DBP of 10mmHg or more being 90% and the normalization rate, deficed as DBP lowering to 90mmHg or below, being 70%. Heart rate, hematology and blood chemistry including blood sugar and lipids were not changed significantly after treatment with isradipine. No significantl side effect was observed except 2 cases of mild transient facial flushing and nausea during the treatment, so could proceed the trial without drug discontinuation in all 30 cases. The results suggest that isradipine is one of the useful and safe drugs in the treatment of mild to moderate essential hypertension.
Blood Glucose ; Blood Pressure ; Chemistry ; Female ; Flushing ; Heart Rate ; Hematology ; Humans ; Hypertension* ; Isradipine* ; Nausea