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*

Vitiligo is a common skin depigmentation disorder and treatment options are generally unsatisfactory and difficult. We report a case of vitiligo resistant to classic treatment showing remarkable repigmentation with topical tacrolimus ointment 0.03%.

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

Detection of cystic lesions in the pancreas has increased because of the widespread use of high-resolution diagnostic imaging techniques. Therefore, cystic lesions of the pancreas constitute an increasingly important category with a challenging differential diagnosis. Squamoid cyst of pancreatic ducts is a recently recognized type of cystic lesion in the pancreas in which cystically dilated ducts are lined by non-keratinized squamous epithelium. Although it is clinically known as benign cystic lesion, we experienced its malignant behavior and report here with review of the international literatures.
Diagnosis, Differential ; Diagnostic Imaging ; Epithelium ; Pancreas ; Pancreatic Cyst ; Pancreatic Ducts* ; Pancreatic Neoplasms

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

PURPOSE: Toll-like receptor 3 (TLR3) recognizes to viral double-stranded RNA and is involved in antiviral defenses. A probable role of TLR3 gene variants in the pathogenesis of aspirin-intolerant asthma (AIA) has been suggested. AIA patients present more frequent asthma exacerbations in which respiratory viral infections could be an exacerbating factor. IgG subclass deficiency was commonly present with bronchial asthma. Based on previous findings, we investigated whether TLR3 variants could affect IgG3 subclass deficiency in AIA. METHODS: We enrolled 279 AIA patients, 403 aspirin-tolerant asthma (ATA) patients, and 315 normal healthy controls (NC) in this study. TLR3 polymorphism at the promoter region -299698G>T was genotyped. The serum levels of IgG subclasses were determined by the single radial immunodiffusion method. Expressions of IgG3 and TLR3 on Epstein-Barr virus transformed-B cells isolated from asthmatic patients were evaluated by flow cytometry to investigate B-cell functions. RESULTS: The TLR3 -299698 T allele was significantly associated with severity and IgG3 deficiency in the AIA group (P=0.044 and P=0.010, respectively), but not in the ATA group. IgG3 expression on B cells from asthmatics with IgG3 deficiency was significantly lower compared to those without (P=0.025). There was a positive correlation between IgG3 expression levels on B cells and serum IgG3 levels (r 2=0.434, P=0.002). CONCLUSION: These results suggest that the TLR3 -299698G>T polymorphism may be associated with IgG3 subclass deficiency and severity in AIA.
Alleles ; Asthma* ; B-Lymphocytes ; Flow Cytometry ; Herpesvirus 4, Human ; Humans ; Immunodiffusion ; Immunoglobulin G* ; Methods ; Polymorphism, Genetic ; Promoter Regions, Genetic ; RNA, Double-Stranded ; Toll-Like Receptor 3

Multiple simultaneous cranial neuropathies occur rarely in diabetes patients. In general, diabetic cranial neuropathy presents in an isolated form and frequently involves oculomotor or facial nerves. We report a 73-year-old man with known type 2 diabetes mellitus who presented with severe dizziness, diplopia and third, fourth and sixth nerve ophthalmoplegia of both eyes. Radiological, laboratory and ophthalmic work-up including magnetic resonance imaging and angiography (MRI and MRA) revealed no specific tumor, aneurysm, or inflammation findings, except for a previous cerebral infarction and atherosclerotic changes in the internal carotid and vertebral arteries. After strict blood glucose control, the multiple cranial nerve palsies spontaneously resolved in 12 weeks. We report the case with a review of the literature.
Abducens Nerve* ; Aged ; Aneurysm ; Angiography ; Blood Glucose ; Cerebral Infarction ; Cranial Nerve Diseases ; Diabetes Mellitus ; Diabetes Mellitus, Type 2 ; Diabetic Neuropathies* ; Diplopia ; Dizziness ; Facial Nerve ; Humans ; Inflammation ; Magnetic Resonance Imaging ; Ophthalmoplegia ; Vertebral Artery

Ankylosing spondylitis (AS) is a systemic inflammatory disorder that affects the axial skeleton. It often involves the extra-articular organs. Cardiovascular involvement is one of the extra-articular manifestations, which is mostly represented by aortic root, valvular heart disease, and conduction disturbances. An aortic sclerosing inflammatory process induces aortic root thickening and rigidity. An aortic aneurysmal change is a rare complication that often leads to life threatening conditions. A few cases regarding aortic aneurysm have been reported, but there are no reported cases in Korea. We report the first case of descending thoracic and abdominal aortic aneurysm in a patient with ankylosing spondylitis.
Aortic Aneurysm ; Aortic Aneurysm, Abdominal* ; Heart Valve Diseases ; Humans ; Korea ; Skeleton ; Spondylitis, Ankylosing*