Hantaviruses are negative-strand RNA viruses that contain three segmented genome and belong to the family Bunyaviridae. Due to such an unique structure of segmented RNA genome, these viruses have a possibility to produce reassortant that have genomic sets mixed with different segments originated from both parental virus during the genetic interaction. To investigate whether this phenomenon occurs actually, Hantaan virus (HTN) could be coinfected with Seoul virus (SEO) to Vero-E6 cell. And also, HTN could be coinfected with Prospect hill virus (PH) to investigate the rate of genetic reassortment according to the genetic distance among the HTN, SEO and PH viruses. As a available method to differentiate the reassortant, we used the multiplex RT-PCR that detect and differentiate the serotype of hantaviruses in mixed infection. Each progeny clone could be screened by multiplex RT-PCR. So, we have constructed the multiplex RT-PCR system that separated amplifications for each segment of Hantaviruses. Our multiplex RT-PCR system provide sensitive, specific and simplified tools for the rapid differentiation of hantaviruses serotypes and also the diagnosis of Hantavirus infections.
Bunyaviridae ; Clone Cells ; Coinfection ; Diagnosis ; Genome ; Hantaan virus ; Hantavirus Infections ; Hantavirus* ; Humans ; Hydrogen-Ion Concentration ; Parents ; RNA ; RNA Viruses ; Seoul virus ; Serotyping*

Prospect Hill virus (PHV) is related antigenically but distinct to Hantaan virus. As a member of genus Hantavirus, PHV has three segmented RNA genome. Among these segments, Small segment(S) encodes nucleocapsid protein (NP) as structural protein and also may do functional nonstructural protein(NSs). We performed in vitro transcription to produce vRNA of PHV S genome. For the first step of in vitro transcription of S genome of PHV, the S RNA segrnent which is 1,675 nucleotides long was amplified by RT-PCR using PCR primers built according to cDNA sequence of PHV S genome. Next, a new PCR primer appended above downstream primer to T7 phage promoter sequence was reconstructed to obtain PCR product containing T7 promoter. Then another PCR was performed. Using this PCR product as the template, in vitro run-off transcription without cloning by transcriptional vector was performed to obtain viral- sense RNA transcript. Thereafter, the size of transcript was assessed by running on formaldehyde agarose gels. Since the transcription reactants contain a-S UTP, the transcript is detectable by autoradiography. The transcript was also detectable by northern hybridization with a-P dCTP- labelled PHV amplicon probe (319 bp) and the initiation start point of run-off transcription was also determined by primer extention analysis. Our data indicate that the in vitro transcript could be produced from the PCR product amplified by PCR primer containing T7 phage promoter without cloning into a phage transcription vector.
Autoradiography ; Bacteriophage T7 ; Bacteriophages ; Clone Cells ; Cloning, Organism ; DNA, Complementary ; Formaldehyde ; Gels ; Genome* ; Hantaan virus ; Hantavirus ; Nucleocapsid Proteins ; Nucleotides ; Polymerase Chain Reaction ; RNA ; Running ; Sepharose ; Uridine Triphosphate ; Viruses*

No abstract available.
Anus, Imperforate* ; Perineum* ; Ultrasonography*

Recently, the proportion of cardiovascular disease as a cause of death in diabetics is in ever increasing tendency. Present study is designed to evaluate the change of myocardial performance by the effect of microangiopathy of diabetics. The measurements of systolic time intervals were obtained from simultaneous high speed recordings(100mm/sec) of electrocardiography, phonocardiography, and carotid pulse tracing in diabetics(male 26, female 31). 1. As the diabetic retinopathy got severe, shortening of LVET and prologation of PEP were noted, but QS2 showed no significant change. 2. PEP/LVET ratio fo diabetics without diabetic retinopathy was significantly higher than that of normal control(p<0.01). 3. PEP/LVET ratio of diabetics with diastolic blood pressure under 95mmHg was significantly higher than that of normal control group(p<0.01), and PEP/LVET ratio of diabetics with diastolic blood pressure over 96mmHg was significantly higher than that of normal control and that of diabetics with diastolic blood pressure under 95mmHg(p<0.01). 4. PEP/LVET ratio of diabetics with proliperative retinopathy was significantly higher than that of normal control and that of diabetics with mild diabetic retinopathy(p<0.01), but this result was not conclusive because of the small case number.
Blood Pressure ; Cardiovascular Diseases ; Cause of Death ; Diabetic Retinopathy ; Electrocardiography ; Female ; Humans ; Phonocardiography ; Systole*

No abstract available.
Constriction, Pathologic* ; Diagnosis*

Current treatment strategies for levodopa-induced psychosis in advanced Parkinson's disease have had limited success. Reduction or discontinuation of levodopa and coadministration with dopamine-blocking neuroleptics may attenuate the psychotic symptoms, but these strategies are associated with worsening of parkinsonian symptoms. Administration of 5-HT3 receptor antagonist ; ondansetron, a newer strategy to attenuate psychosis of Parkinson' disease without motor deterioration was introduced. A 41-year-old young-onset male, who was diagnosed as Parkinson's disease 7 years ago, was treated with levodopa therapy, and had levodopa-induced psychosis(delusion, hallucination, paranoid, insomnia). After trial of ondansetron, he showed improvement in the Brief Psychiatric Rating Scale(from 21 points to 9 points) in spite of increasing the dosage of levodopa. With ondansetron, we could increase the dosage of levodopa without psychotic complications(esp, hallucination), and he showed improvement in the motor fluctuation.
Adult ; Antipsychotic Agents ; Hallucinations ; Humans ; Levodopa ; Male ; Ondansetron ; Parkinson Disease ; Psychotic Disorders* ; Receptors, Serotonin, 5-HT3

No abstract available.
Child* ; Humans ; Thyroid Gland*

Dentocutaneous fistula of the face neck is an infrequent manifestation of chronic dental infection. A periapical dental abscess may be initiated by disease, trauma, or thermal or chemical injury and develops into an extensive necrosis of surrounding tissue. Diagnostic errors can result in multiple excision, biopsies, and ineffective long-term antibiotic therapy. Awarness that periapical dental abscess is the most common etilolgic factor of cutaneous sinus tracts involves the face & neck will facilitate their early diagnosis and prevent needless treatment or anxiety for the patient. From 1994 to 1998, we have performed 6 cases of dentocutaneous fistula. Among of them, three were men and the others were women, age ranged from 18 to 66-year-old, and morbidity period was from 3 weeks to 3 years, the follow-up period ranged from 5 to 18 months (mean period 11 months). In conclusion, an understanding of the pathogenesis of cutaneous fistulae arising from dental infections will lead to proper early diagnosis and treatment without unnecessary surgery.
Abscess ; Aged ; Anxiety ; Biopsy ; Cutaneous Fistula ; Diagnostic Errors ; Early Diagnosis ; Female ; Fistula* ; Follow-Up Studies ; Humans ; Male ; Neck ; Necrosis ; Unnecessary Procedures

Adenosine deaminase(ADA)has been used for the diagnosis of localized tuberculosis.The titer of ADA was measured from the ocular fluid of the patients who had undergone vitreoretinal surgery due to non-tuberculous vitreoretinal disease. Forty five patients were included and twenty patients were male. Mean age was 51.9 years. Four patients had diabetes mellitus and two had essential hypertension. The most frequent cause of vitreoretinal surgery was proliferative vitreoretinopathy(12 eyes). For the analysis of ADA titer, vitreous fluid(group I)was aspirated in 36 patients undergoing pars plana vitrectomy and subretinal fluid(group II)in 9 patients undergoing scleral encircling. Mean titer of ADA was 9.01+/-15.69 IU/L, and there was no significant difference on the statistics between group I(6.06+/-13.75 IU/L)and group II(20.83+/-18.17 IU/L). All 45 eyes showed negative reaction to polymerase chain againt tuberculosis, and 25 of 29 patients had positive reaction to tuberculin skin test. There was no statistically significant relationship between ADA titer and tuberculin skin test. This result suggests that ADA may be used for the diagnosis of posterior ocular tuberculosis, since the normal concentration in the posterior ocular fluid was compatible with the one in other body fluids where it has been used for the diagnosis of localized tuberculosis.
Adenosine Deaminase* ; Adenosine* ; Body Fluids ; Diabetes Mellitus ; Diagnosis ; Humans ; Hypertension ; Male ; Skin Tests ; Tuberculin ; Tuberculosis ; Tuberculosis, Ocular ; Vitrectomy ; Vitreoretinal Surgery

Hantaan virus is the causative agent of rather severe form of hemorrhagic fever with renal syndrome which occurs widely in north-eastern Asia including Korea, China and far eastcrn part of Russia. Although several types of vaccine for this disease have been developed, the therapeutic agent has not been developed yet. Therefore, we launched the construction of ribozyme to be used as the therapeutic purpose of the disease. Ribozyme which cleaves RNA as an enzyme is a RNA oligonucleotide specific to target RNA. We constructed a ribozyme oligonucleotide aimed at S genomic RNA segment of Hantaan virus (strain 76-118) containing T7 promoter region cornplementary to promoter primer oligonucleotide. Then two oligonucleotides were annealed to prepare double stranded transcription template, and transcription was performed in vitro. Thus, we could prepare the clone of whole S segment of the virus by RT-PCR, and then BamHI/HinCII fragment of the S genome segment was subcloned to pT7T319U vector containing T7 promoter in genome sense. The substrate transcript was made by run-off transcription. These substrate and ribozyme transcripts were used to detect cleavage activity of the ribozyme to the target RNA substrate prior to its application to cultured cell. The cleavage reaction showed that the ribozyme cleaves the target RNA which is S segment of Hantaan virus. To know whether the ribozyme works in cell infected with Hantaan virus as well, the ribozyme was transfected to Vero-E6 cell by lipofectin after inoculation of the virus. The transfected ribozyme was detectable in the cell by RT-PCR utilizing ribozyme specific primers. On 7 days after inoculation, the culture media were harvested and used to determinate viral titers by immunoenzyme plaque assay. In contrast to the mock transfected negative control, the viral titers of the cultures transfected at 1, 2 and 3 days after the virus inoculation were lowered to 1/100 level. This result suggests that the ribozyme inhibits the multiplication of Hantaan virus in cultured cell successfully in early stage of infection, and ribozyme is a possible new anti-viral drug against the virus infection.
Asia ; Cells, Cultured ; China ; Clone Cells ; Culture Media ; Genome ; Hantaan virus* ; Hemorrhagic Fever with Renal Syndrome ; Korea ; Oligonucleotides ; Promoter Regions, Genetic ; RNA ; Russia