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.
Lung Neoplasms* ; Lung* ; Pneumothorax, Artificial*

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

Some structures should be noted in the modified Stoppa approach for the treatment of acetabular fractures. Among them, the external iliac vessels should be protected with special care during surgery. If there is any history of previous abdominal surgery or a delay in surgery, it is predicted that the intra-abdominal adhesion will be severe. In this case, thorough preparation and caution are required before surgery because a dangerous situation may arise due to blood vessel damage during surgery. On the other hand, there are no specific reports on these cases. The authors report a case of severe adhesion of the external iliac vessels with a history of previous abdominal surgery.

The Rieger's syndrome is characterized by the ocular anomalies include posterior embryotoxon(a prominent Schwalbe's line), tissue strands across the anterior chamber angle and variable degrees of iridic distortion and systemic defects most often involve the teeth and facial bones. These disorders are typically bilateral, are usually diagnosed at birth or in early childhood, and most have genetic basis of autosomal dominant inheritance. A large number of patients have glaucoma due to developmental defects of the anterior chamber angle structures. The authors experienced two cases of Rieger's syndrome which were occurred in a 13-year-old female in one family and in a 21 year-old male in the other family who had characteristic ocular and other systemic abnormalities associated with developmental glaucoma.
Adolescent ; Anterior Chamber ; Facial Bones ; Female ; Glaucoma ; Humans ; Male ; Parturition ; Tooth ; Wills ; Young Adult

BACKGROUND: The present study was carried out in association with neutrophilic respiratory burst in the lung in order to clarify the pathogenesis of acute respiratory distress syndrome(ARDS) following acute severe hemorrhage. Because oxidative stress has been suggested as one of the principal factors causing tissue injury, the role of free radicals from neutrophils was assessed in acute hemorrhage-induced lung injury. METHOD: In Sprague-Dawley rats, hemorrhagic shock was induced by withdrawing blood(20 ml/kg of B.W) for 5 min and the hypotensive state was sustained for 60 min. To determine the mechanism and role of oxidative stress associated with phospholipase A2(PLA2) by neutrophils, the level of lung leakage, pulmonary myeloperoxidase(MPO), and the pulmonary PLA2 were measured. In addition, the production of free radicals was assessed in isolated neutrophils by cytochemical electron microscopy in the lung. RESULTS: In hypotensive shock-induced acute lung injury, the pulmonary MPO, the level of lung leakage and the production of free radicals were higher. The inhibition of PLA2 with mepacrine decreased the pulmonary MPO, level of lung leakage and the production of free radicals from neutrophils. CONCLUSION: A. neutrophilic respiratory burst is responsible for the oxidative stress causing acute lung injury followed by acute, severe hemorrhage. PLA2 activation is the principal cause of this oxidative stress.
Acute Lung Injury* ; Free Radicals ; Hemorrhage ; Lung ; Lung Injury ; Microscopy, Electron ; Neutrophils* ; Oxidative Stress ; Phospholipases A2* ; Phospholipases* ; Quinacrine ; Rats, Sprague-Dawley ; Respiratory Burst* ; Shock, Hemorrhagic

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

No abstract available.
Idiopathic Pulmonary Fibrosis*