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*

No abstract available.
Congenital Hypothyroidism*

Generally, the hemolytic disease of newborn caused by anti-c is characterized by the abnormal titer of anti-c in mother's serum and neonatal progressive jaundice caused by IgG induced hemolytic anemia. A female neonate delivered by cesarean section was admitted to Hangang Sacred Hospital due to severe jaundice and apnea on Jul. 7 1998. On admission, laboratory work up was revealed immune hemolytic anemia, but no incompatibility of ABO and Rh(D) system between mother and baby. Minor blood groups of mother and this second baby was ce and Cce. The serologic examination showed positive direct and indirect Coombs' test and the cause of jaundice was proved as the result of hemolytic disease due to anti-c via antibody screening and identification. After 2 times of exchange transfusion, the jaundice was subsided. So we reported a case of hemolytic disease due to anti-c with review of previous literatures.
Anemia, Hemolytic ; Apnea ; Blood Group Antigens ; Cesarean Section ; Coombs Test ; Erythroblastosis, Fetal* ; Female ; Humans ; Immunoglobulin G ; Infant, Newborn ; Jaundice ; Mass Screening ; Mothers ; Pregnancy

No abstract available.
Carcinoma, Squamous Cell* ; Teratoma*

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

PURPOSE: To analyze the characteristies of traumatic brainstem injury by CT and MR MATERIALS AND METHODS: CT and MR studies of 10 patients with traumatic brainstem lesion in MR were retrospectively reviewed, particularly attended to location, signal intensity and associated lesions. RESULTS: CT failed to depict 8 of 10 brainstem lesions. All lesions were detected in MR images with T2-weighted images showing higher detection rate (n=10) (100%) than Tl-weighted images (n=3) (30%) or CT (n=2) (20%). The brainstem lesions located in the dorsolateral aspects of the rostral brainstem(mid brain and upper pons)in 7 (70%) cases, in ventral aspects of rostral brain in 2 (20%) cases and in median portion of pons in 1 (10%) case. Corpus callosal (n=5), Iobar white matter(n=5) diffuse axonal injury, and 2 hemorrhagic lesions in basal ganglia were the associated findings. CONCLUSION: MR imaging is more helpful than CT in the detection of brainstem injury, especially T2 weighted images. Primary brainstem lesions were typically located in the dorsolateral aspect of rostral brainstem(midbrain and upper pons). Corpus callosum and white matter lesions were frequently associated.
Basal Ganglia ; Brain ; Brain Stem* ; Corpus Callosum ; Diffuse Axonal Injury ; Humans ; Magnetic Resonance Imaging ; Pons ; Retrospective Studies

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

OBJECTIVES: The purpose of this study is to evaluate effects of patients' age, amenorrhea duration, motivation, purpose, regimen, side effects on compliance to hormone replacement therapy(HRT) Methods: Compliance to HRT was assessed in 258 postmenopausal women who began HRT in the Menopause Clinic of Hangang Sacred Heart Hospital, Hallym university. One hundred sixty nine women who had intact uterus received continuous combined therapy with 0.625 mg of conjugated equine estrogen (CEE) and 100 mg of micronized progesterone. Eighty nine women who were hysterectomized received only 0.625 mg of CEE. RESULTS: A total of 103 women (39.9%) reported that they continued HRT at least for one year. Continuation of HRT was significantly more common among women who started HRT becuase of recommendation by gynecologist after hysterectomy in our department (p< 0.05). A greater percentage of women who received only CEE without uterus continued HRT than women who received continuous combined therapy (p< 0.05). Duration of amenorrhea, reasons for taking HRT and side effects such as irregular bleeding or breast tenderness were not related to the continuation of HRT. CONCLUSION: Education of menopausal women about the benefit/risk and side effects of HRT may be critical in improving compliance.
Amenorrhea ; Breast ; Compliance* ; Education ; Estrogens ; Female ; Heart ; Hemorrhage ; Hormone Replacement Therapy ; Humans ; Hysterectomy ; Menopause ; Motivation ; Progesterone ; Uterus

Since Thomas et al reported the first instance of successful removal of a broken intravascular guidewire without surgical intervention by using a bronchoscopic forceps in 1964, there have been a number of nonsurgical removal of intravascular or intracardiac foreign bodies using the bronchoscopic forceps, loop snare or basket stone catchers. We experienced a successful retrieval of accidentally broken subclavian puncture catheter from right atrium in 76 year old male patient with cerebral infarction admitted to this hospital on May, 1989. By percutaneous catheterization via right basilic vein, using a loop snare which is modified manually by guidewire for PTCA(USIC(R) Teflon coated PTCA Guide wire : ".014"), we removed successfully foreign body from right atrium. Therefore, we report a case of nonsurgical retrieval of foreign body from right atrium with the review of the literature.
Aged ; Catheterization ; Catheters ; Cerebral Infarction ; Foreign Bodies* ; Heart Atria ; Humans ; Male ; Ocimum basilicum ; Polytetrafluoroethylene ; Punctures ; SNARE Proteins ; Surgical Instruments ; Veins