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*

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*

Takayasu's arteritis is an arteritis of undetermined etiology, which affects the aorta, the proximal portions of its major branches, and the pulmonary arteries, and causes coarctation, occlusion, or aneurysmal dilatation of the affected vessels. Authors has reported 9 cases in 1973, and another 24 cases in 1977. Thereafter, during next 4 years, authors experienced another 42 cases and obtained some additional results. The results are as follows: 1. Among 75 cases, 10 are male and 65 female patients with sex ratio of 1:6.5, and about 2/3 of total patients are under 30 years of age. 2. Headache, dizziness, absent or weak radial pulse and hypertension are the most common symptoms and signs. 3. Conventional chest roentgenography may be helpful but not specific in diagnosis of Takayasu's arteritis. 4. The aortographic findings are characteristic and pathognomonic in diagnosis of Takayasu's arteritis. In our series, the most common findings are stenosis or occlusion of subclavian artery, diffuse narrowing and aneurysmal dilatation of abdominal aorta and its branches, narrowing with irregular contour of descending thoracic aorta, and renal artery involvement. 5. Involvement of the aorta was classified as extensive type in 38 cases, descending thoracic and abdominal type in 22 cases and arch type in 15 cases. 6. As total aortography in cluding abdominal aorta uncovers evidence of unsuspected involvement of aorta and its branches, it is of paramount importance in the diagnosis of Takayasu's arteritis.
Aneurysm ; Aorta ; Aorta, Abdominal ; Aorta, Thoracic ; Aortography ; Arteritis ; Constriction, Pathologic ; Diagnosis ; Dilatation ; Dizziness ; Female ; Headache ; Humans ; Hypertension ; Male ; Pulmonary Artery ; Radiography ; Renal Artery ; Sex Ratio ; Subclavian Artery ; Takayasu Arteritis* ; Thorax

Percutaneous transhepatic biliary drainage was successfully made 20 times on 17 patients of obstructivejaundice for recent 1 year since June 1981 at department of radiology in Seoul National University Hospital. Thecauses of obstructive jaundice was CBD Ca in 13 cases, metastasis in 2 cases, pancreatic cancer in 1 case and CBDstone in 1 case. Percutaneous transhepatic biliary drainage is a relatively easy, safe and effective method whichcan be done after PTC by radiologist. It is expected that percutaneous transhepatic biliary drainage should bedone as an essential procedure for transient or permanent palliation of obstructive jaundice.
Drainage ; Humans ; Jaundice, Obstructive ; Methods ; Neoplasm Metastasis ; Pancreatic Neoplasms ; Seoul

Transglutaminase(TGase) is a calcium dependent enzyme that catalyse and acyl transfer reaction forming epsilon-(gamma-glutamyl)-lysine cross linkage. the major known effect of TGase is its important role in the programmed cell death manifested in the granular layer of the skin and acidophilic bodies in the viral hepatitis and neoplastic processes. The enzyme activity, immunohistochemical reaction using polyclonal antibodies against cytosolic TGase C, light and electron microscopic studies and TdT staining of the transplanted fibrosarcoma cells in C3H mouse with radiation therapy were done. The presence of TGase was detected immunohistochemically by avidin-biotin peroxidase complex (ABC) method Apoptosis were significantly induced after irradiation dependent with time factors and irradiation doses, resulted in marked and confluent tumor cell loss. Highest activity of the cytosolic form of TGase was noted at 24 hours and decrease after then while membrane bounded form of the TGase showed no significant changes. Immunohistochemical staining revealed strong positive reaction in the sarcoma cells in diffuse fasion and around the necrotic foci in the cytoplasm. Terminal dideoxynucleotidyl transferase(TdT) staining revealed increasing numbers of apotptic cells from two hours after irradiation. In the mechanism of decreasing tumor size and cell death in radiation therapy, apoptosis plays an important role and during that process transglutaminse might do some irreversible cross-linking effects of cytoplasmic proteins causing cell death in part.
Mice ; Animals

No abstract available.
Dental Enamel* ; Lasers, Solid-State*

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

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

Two cases of exercise-induced atrial fibrillation were diagnosed in 68 years old female and 47 years old male. They complained palpitation during exercise, and emotional upset for the last 5 years. There was no evidence of organic heart disease except mild hypertension in female patient. The atrial fibrillation was induced repeatedly by treadmill exercise test and intravenous infusion of isoproterenol. The beta blocker(atenolol 50mg po dialy) prevented the induction of atrial fibrillation during treadmill exercise. We report two cases of exercise-induced atrial fibrillation which is very rare and might be related to catecholamine.
Aged ; Atrial Fibrillation* ; Exercise Test ; Female ; Heart Diseases ; Humans ; Hypertension ; Infusions, Intravenous ; Isoproterenol ; Male ; Middle Aged

To assess the likelihood of procedural success in patients with multivessel coronary artery disease, 46 consecutive patients (male 34, female 12, mean age 60+/-9 years) umderwent single or multiple site angioplasty. The clinical diagnosis of unstable angina was in 20(44%), stable angina in 10 and acute or old myocardial infarction in 16. Coronary angiographic findings of 2 vessel disease was in 38(83%), triple vessel disease in 8. Left ventricular function was generally well preserved (mean ejection fraction 65+/-12%, range 30-82%) and mean 2.0 stenosis per patient angic, lasty had attempted. Single vessel angioplasty (SVA) was performed in 13 and multivessel angioplasty (MVA) in 33. Procedural success was achieved in 79(86%) out of total 92 stenoses. Sixty-six(88%) out of 75 stenoses in MVA and 13(76%) out of 17 stenoses in SVA had procedural success respectively. According to angiographic morphology of lesions, procedural success of type A stenoses was 17/17(100%), type B stenoses 57/66(86%) and type C stenoses was 5/9(33%). In 13 failures included inability to pass the guide wire cross the lesion in 7, inability to guide the griding catheter in 2 and inability to dilate lesions in 4. Before and after angioplasty, treadmill test (modified Bruce protocol)could be performed in 29 patients. Total duration of exercise and maximal double product improved significantly from 8.5+/-2.3 minute 5188+/-2403 to 12.2+1.3 min., 23,062+/-4111 respectively (p<0.001). 17 out of 24 patients who had positive treadmill test before angioplasty showed negative conversion after procedure. Complications included dissection in 29, prolongd chest pain in 5, acute closure in 3, cardiac tamponade in 1 and ventricular fibrillation due to side branch oclusion in 1. Thus, coronary angioplasty in selected paients with multivessel coronary artery disease might be useful and have relatively good immediate results, but the long-term efficacies with other forms of treatment must be evaluated prospectively.
Angina, Stable ; Angina, Unstable ; Angioplasty* ; Cardiac Tamponade ; Catheters ; Chest Pain ; Constriction, Pathologic ; Coronary Artery Disease* ; Coronary Vessels* ; Diagnosis ; Exercise Test ; Female ; Humans ; Myocardial Infarction ; Ventricular Fibrillation ; Ventricular Function, Left