BACKGROUND: The determination of serotype of enteroviruses is useful for the discrimination between sporadic and epidemic infections. The conventional serotyping method is time-consuming and labor-intensive. Recently, molecular method was introduced for the serotyping of enteroviruses. The aim of this study was to establish a method to isolate and analyze enteroviruses from various specimens utilizing molecular biological techniques and to determine which strains were phylogenetically related to clinical samples. METHODS: Clinical samples in this study included 164 cerebrospinal fluid (CSF), 136 stool, 15 sera, 6 throat swab, 5 urine, and 4 sputa, which were obtained from hospitalized patients, primarily infants or children presenting symptoms of aseptic meningitis in 1998. RD cells were used for enterovirus isolation. RT-PCR was performed with RD cell lysate showing CPE. The primers 011 and 012 were used for the VP1 region, and the primers EN1 and EN2 for 5'-UTR. The nucleotide sequences of VP1 region were determined and analyzed with BLAST program. RESULTS: Among 333 samples, only 23 samples produced CPE: 17 samples at first and six samples at the second blind passage. Fifteen isolates were related to coxsackievirus B2 two to echovirus 4, three to echovirus 6, and three to echovirus 18. All 23 viral isolates displayed a nucleotide sequence identity of 80-95%, compared with the reference serotypes. However, the identity was increased up to 93-100% when the VP1 region was translated into amino acids CONCLUSIONS: Since CB2 type was 55% among enteroviral isolates, the CB2 was determined as the major causative serotype of enteroviral meningitis in 1998. CB2 type was emerged between June and July, EC4 and EC6 was limited to July, and EC18 was in August.
Amino Acids ; Base Sequence ; Cerebrospinal Fluid ; Child ; Discrimination (Psychology) ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus* ; Humans ; Infant ; Meningitis* ; Meningitis, Aseptic ; Pharynx ; Serotyping*

BACKGROUND: The determination of serotype of enteroviruses is useful for the discrimination between sporadic and epidemic infections. The conventional serotyping method is time-consuming and labor-intensive. Recently, molecular method was introduced for the serotyping of enteroviruses. The aim of this study was to establish a method to isolate and analyze enteroviruses from various specimens utilizing molecular biological techniques and to determine which strains were phylogenetically related to clinical samples. METHODS: Clinical samples in this study included 164 cerebrospinal fluid (CSF), 136 stool, 15 sera, 6 throat swab, 5 urine, and 4 sputa, which were obtained from hospitalized patients, primarily infants or children presenting symptoms of aseptic meningitis in 1998. RD cells were used for enterovirus isolation. RT-PCR was performed with RD cell lysate showing CPE. The primers 011 and 012 were used for the VP1 region, and the primers EN1 and EN2 for 5'-UTR. The nucleotide sequences of VP1 region were determined and analyzed with BLAST program. RESULTS: Among 333 samples, only 23 samples produced CPE: 17 samples at first and six samples at the second blind passage. Fifteen isolates were related to coxsackievirus B2 two to echovirus 4, three to echovirus 6, and three to echovirus 18. All 23 viral isolates displayed a nucleotide sequence identity of 80-95%, compared with the reference serotypes. However, the identity was increased up to 93-100% when the VP1 region was translated into amino acids CONCLUSIONS: Since CB2 type was 55% among enteroviral isolates, the CB2 was determined as the major causative serotype of enteroviral meningitis in 1998. CB2 type was emerged between June and July, EC4 and EC6 was limited to July, and EC18 was in August.
Amino Acids ; Base Sequence ; Cerebrospinal Fluid ; Child ; Discrimination (Psychology) ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus* ; Humans ; Infant ; Meningitis* ; Meningitis, Aseptic ; Pharynx ; Serotyping*

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) was found in several serotypes of E coli including 0157 serotype. Sorbitol-MacConkey agar: may be useful for the detection of E. coli 0157, but is not helpful for the detection of sorbitol-fermenting STEC other than 0157. Moreover, some strains of E. colt 0157 can ferment sorbitol. In this study, in situ hybridization using DNA probe of shiga toxin was used for the isolation of STEC from the PCR-positive stool and -Sequenbe analysis of a part of shiga toxin gene was performed. METHODS: The stool was incubated in LB broth overnight and DNA was extracted from the culture fluid. Multiplex PCR was performed with primers for stxl and stx2 genes. Specimen showed PCR-positive was incubated on MacConkey agar and colonies were blotted with nitrocellulose membrane. Digoxigenin-labelled DNA probe for shiga toxin was made by PCR and the positive colonies were detected with anti-digoxigenin-alkaline phosphatase conjugate and nitroblue tetrazolium. Agglutination test with antisera was performed for the serotying and VTEC-RPLA kit was used for the toxin production. Sequence analysis of PCR products was performed with automatic sequence analyser. RESULTS: An stxl-negative, but stx2-positive PCR was observed in a three-year-old girl, who visited Kumi Hospital on July 19, 1999 complaining of vomiting and diarrhea. The positive colonies were isolated by in situ hybridization using stx2-specific DNA probe. The titers of stxl and stx2 by VTEC-RPLA test were negative and 1:64, respectively. Agglutination for the serotyping was not observed with all of the 0 antisera. 160-nucleotide sequence of stx2 of this isolate was identical with bacteriophage 933W (GenBank X07865), except for the change (T-C) of 957th nucleotide and amino acid sequence was identical each other. CONCLUSIONS: For the sensitive detection of STEC from the stool of patients with diarrhea, multiplex PCR is recommended with stxl- and stx2-specific primers. And in situ hybridization should be performed in PCR-positive specimen for the isolation of STEC. This method may be helpful for the detection of STEC as the causative microorganisms in food-borne outbreak.
Agar ; Agglutination ; Agglutination Tests ; Amino Acid Sequence ; Bacteriophages ; Collodion ; Diarrhea ; DNA ; Escherichia coli ; Female ; Gyeongsangbuk-do ; Humans ; Immune Sera ; In Situ Hybridization ; Membranes ; Multiplex Polymerase Chain Reaction ; Nitroblue Tetrazolium ; Polymerase Chain Reaction ; Sequence Analysis* ; Serotyping ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli* ; Sorbitol ; Vomiting

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) was found in several serotypes of E coli including 0157 serotype. Sorbitol-MacConkey agar: may be useful for the detection of E. coli 0157, but is not helpful for the detection of sorbitol-fermenting STEC other than 0157. Moreover, some strains of E. colt 0157 can ferment sorbitol. In this study, in situ hybridization using DNA probe of shiga toxin was used for the isolation of STEC from the PCR-positive stool and -Sequenbe analysis of a part of shiga toxin gene was performed. METHODS: The stool was incubated in LB broth overnight and DNA was extracted from the culture fluid. Multiplex PCR was performed with primers for stxl and stx2 genes. Specimen showed PCR-positive was incubated on MacConkey agar and colonies were blotted with nitrocellulose membrane. Digoxigenin-labelled DNA probe for shiga toxin was made by PCR and the positive colonies were detected with anti-digoxigenin-alkaline phosphatase conjugate and nitroblue tetrazolium. Agglutination test with antisera was performed for the serotying and VTEC-RPLA kit was used for the toxin production. Sequence analysis of PCR products was performed with automatic sequence analyser. RESULTS: An stxl-negative, but stx2-positive PCR was observed in a three-year-old girl, who visited Kumi Hospital on July 19, 1999 complaining of vomiting and diarrhea. The positive colonies were isolated by in situ hybridization using stx2-specific DNA probe. The titers of stxl and stx2 by VTEC-RPLA test were negative and 1:64, respectively. Agglutination for the serotyping was not observed with all of the 0 antisera. 160-nucleotide sequence of stx2 of this isolate was identical with bacteriophage 933W (GenBank X07865), except for the change (T-C) of 957th nucleotide and amino acid sequence was identical each other. CONCLUSIONS: For the sensitive detection of STEC from the stool of patients with diarrhea, multiplex PCR is recommended with stxl- and stx2-specific primers. And in situ hybridization should be performed in PCR-positive specimen for the isolation of STEC. This method may be helpful for the detection of STEC as the causative microorganisms in food-borne outbreak.
Agar ; Agglutination ; Agglutination Tests ; Amino Acid Sequence ; Bacteriophages ; Collodion ; Diarrhea ; DNA ; Escherichia coli ; Female ; Gyeongsangbuk-do ; Humans ; Immune Sera ; In Situ Hybridization ; Membranes ; Multiplex Polymerase Chain Reaction ; Nitroblue Tetrazolium ; Polymerase Chain Reaction ; Sequence Analysis* ; Serotyping ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli* ; Sorbitol ; Vomiting

Rectus sheath hematoma (RSH) is a rare condition caused by hemorrhage into the rectus sheath. It is usually associated with severe cough, abdominal surgery, coagulopathy, and anticoagulation treatment. RSH can be difficult to diagnose and can be misdiagnosed as acute appendicitis, as diverticulitis, or as an ovarian mass. Although RSH usually presents as a benign condition, it can be life threatening, especially in the critically-ill patient. Here, we report a case of fatal RSH due to hypovolemic shock in a critically-ill 73-year-old woman, who had received heparin treatment due to acute myocardial infarction in the intensive care unit and who had been successfully treated by conservative management.
Aged ; Appendicitis ; Cough ; Diverticulitis ; Female ; Hematoma ; Hemorrhage ; Heparin ; Humans ; Hypovolemia ; Intensive Care Units ; Myocardial Infarction ; Rectus Abdominis ; Shock

BACKGROUND: The polymerase chain reaction (PCR) test is important for the confirmatory diagnosis of tuberculosis (TB) caused by Mycobacterium tuberculosis. The aim of this study was to analyze the yield of repeated PCR testing in patients with confirmed pulmonary TB. METHODS: The medical records of 130 patients, who had more than two consecutive PCR tests and a M. tuberculosis-positive sputum culture from August, 2006 to December, 2007, were retrospectively reviewed for the purposes of this study. A positive TB-PCR test was defined as at least one positive test result. RESULTS: The cumulative positive PCR test rate was 80% (104/130), with gradually increasing rates of positive findings upon the first, second and third TB-PCR tests with 52.3%, 68.5% and 75.4%, respectively. However, further testing did not increase the positive rate further. CONCLUSION: Repeated PCR testing at least three times for M. tuberculosis is helpful for diagnosis of pulmonary TB.
Humans ; Medical Records ; Mycobacterium tuberculosis ; Polymerase Chain Reaction ; Retrospective Studies ; Sputum ; Tuberculosis ; Tuberculosis, Pulmonary

The presence of epidermal growth factor receptor (EGFR) mutation is a prognostic and predictive marker for EGFR-tyrosine kinase inhibitor (TKI) therapy. However, inevitably, relapse occurs due to the development of acquired resistance, such as T790M mutation. We report a case of repeated responses to EGFR-TKIs in a never-smoked woman with adenocarcinoma. After six cycles of gemcitabine and cisplatin, the patient was treated by gefitinib for 4 months until progression. Following the six cycles of third-line pemetrexed, gefitinib retreatment was initiated and continued with a partial response for 6 months. After progression, she was recruited for an irreversible EGFR inhibitor trial, and the time to progression was 11 months. Although EGFR direct sequencing on the initial diagnostic specimen revealed a wild-type, we performed a rebiopsy from the progressed subcarinal node at the end of the trial. The result of peptide nucleic acid clamping showed L858R/L861Q.
Adenocarcinoma ; Cisplatin ; Constriction ; Deoxycytidine ; Epidermal Growth Factor ; Female ; Glutamates ; Guanine ; Humans ; Lung ; Lung Neoplasms ; Phosphotransferases ; Quinazolines ; Receptor, Epidermal Growth Factor ; Recurrence ; Retreatment ; Pemetrexed

A 5 year-old, intact female Yorkshire terrier was referred for dysuria and dyschezia. The radiographic and ultrasound examination showed a round shaped mass caudal to the urinary bladder that contained anechoic fluid within the thin walls. During surgery, the cyst was noted to be attached to the outer wall of the vagina, not connected to the vaginal lumen. Cystic fluid was removed and the cystic wall was resected. Then the remaining cystic wall was omentalized to prevent a recurrence. Histological examination confirmed that the cyst was of Wolffian duct origin. In this case, a large Gartner duct cyst causing urological problems was diagnosed and removed by surgical resection.
Animals ; Constipation/etiology/veterinary ; Cysts/surgery/ultrasonography/*veterinary ; Dog Diseases/*pathology/surgery/ultrasonography ; Dogs ; Dysuria/etiology/veterinary ; Female ; Treatment Outcome ; Vaginal Diseases/complications/pathology/surgery/*veterinary ; Wolffian Ducts/*pathology/surgery

Pulmonary hypertension is a frequent complication of chronic obstructive pulmonary disease (COPD) and associated with a worse survival and increased risk of hospitalization for exacerbation of COPD. However, little information exists regarding the potential role of systemic inflammation in pulmonary hypertension of COPD. The purpose of the present study was to investigate the degree of C-reactive protein (CRP) and endothelin-1 (ET-1) levels in COPD patient with and without pulmonary hypertension. The levels of CRP and ET-1 were investigated in 58 COPD patient with pulmonary hypertension and 50 patients without pulmonary hypertension. Pulmonary hypertension was defined as a systolic pulmonary artery pressure (Ppa) > or =35 mmHg assessed by Doppler echocardiography. Plasma CRP and ET-1 levels were significantly higher in patients with pulmonary hypertension than in patients without hypertension. There were significant positive correlations between the plasma ET-1 level and CRP level in the whole study groups. For COPD patients, systolic Ppa correlated significantly with plasma CRP levels and plasma ET-1 levels. These findings support a possibility that CRP and ET-1 correlate to pulmonary hypertension in COPD patients.
Aged ; Blood Pressure ; C-Reactive Protein/*analysis ; Echocardiography, Doppler ; Endothelin-1/*blood ; Female ; Humans ; Hypertension, Pulmonary/*blood/complications ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive/*blood/complications

BACKGROUND: As one of harm reduction strategies, tobacco manufacturers have begun to introduce lower-yield cigarettes. Lower-yield cigarettes, so called light cigarettes, have been perceived as less hazardous by some smokers. However, there have been very few studies concerning smoking lower yield products the lead to lower nicotine absorption. We evaluated the association between brand nicotine yield of cigarettes and actual nicotine intake by measuring urinary cotinine. METHODS: Four hundred sixty four male smokers aged 18 or over who participated in health check-ups in a hospital from May to October 2007 filled out a self-administered smoking questionnaire. Urinary cotinine concentration was measured at the time of participation. The subjects were divided into three groups (ultralight [nicotine: 0.05 mg], light [0.1 mg], and regular [> 0.1 mg] group) according to the level of brand nicotine yield of cigarettes which they smoked. RESULTS: The median urinary cotinine concentrations of ultralight (N = 62), light (N = 216), and regular (N = 186) groups were 735.5 ng/mL (interquartile range, 320 to 1,300 ng/mL), 956.0 ng/mL (429 to 1,491 ng/mL), and 1,067.5 ng/mL (615 to 1,613 ng/mL), respectively. There was a signifi cant difference in urinary cotinine between the regular and the other groups (P = 0.015). However, multiple logistic regression analysis to evaluate the risk of being in the highest quartile of urinary cotinine concentration (> or = 1,532 ng/mL) after adjusting for possible confounding variables showed that the odds ratios were 0.84 (95% CI, 0.52 to 1.37) in the light nicotine group and 0.82 (95% CI, 0.38 to1.72) in the ultralight nicotine group compared to the regular nicotine group. CONCLUSION: There was no significant difference in the risk of elevated urinary cotinine concentrations in male adult smokers according to brand nicotine yield of cigarettes groups.
Absorption ; Adult ; Aged ; Androsterone ; Confounding Factors (Epidemiology) ; Cotinine ; Harm Reduction ; Humans ; Light ; Logistic Models ; Male ; Nicotine ; Odds Ratio ; Smoke ; Smoking ; Tobacco ; Tobacco Products