Akabane, Aino and Chuzan virus are arthropod-borne (arbo)viruses mainly associated with reproductive failures in cattle. We investigated apoptosis in Vero cells (C-1586) infected with Akabane, Aino and Chuzan virus. The fragmentation of chromosomal DNA was simultaneously detected with the progress of cytopathic effect from 48 hr to 72 hr post infection, depending on viruses. Although the treatment of cycloheximide blocked apoptosis in Vero cells infected with three viruses, actinomycin D did not prevent DNA oligomerization, thus indicating that de novo viral protein synthesis is critical for viral apoptosis. In addition, the activation of caspase-3 was also detected in Vero cells by indirect fluorescent assay. From the present results, it is of future interest whether apoptotic characteristics of these viruses are related to pathogenecity in vivo.
Animals ; Apoptosis/*physiology ; Bunyaviridae/*physiology ; Caspase 3 ; Caspases/metabolism ; Cercopithecus aethiops ; Cytopathogenic Effect, Viral/*physiology ; DNA Fragmentation/physiology ; Dactinomycin ; Enzyme Activation ; Orbivirus/*physiology ; Vero Cells

BACKGROUND/AIMS: Portal hypertension in cirrhosis is associated with a hyperdynamic circulation, which is characterized by hypervolemia, high cardiac output, arterial hypotension and low peripheral vascular resistance. These circulatory abnormalities are thought to be secondary to a splanchnic arteriolar vasodilation related to the increase in portal pressure. Studies assessing regional hemodynamics in patients of cirrhosis with ascites have shown vasoconstriction in the renal circulation and in peripheral vascular territory. This study was designed to assess the cerebral vascular resistance in cirrhotic patients with ascites. METHODS: The resistive index in the middle cerebral artery and in a renal interlobar artery were measured by Doppler ultrasonography in 12 cirrhotic subjects without ascites, 23 cirrhotic subjects with ascites, and 8 healthy subjects. The arterial blood pressure and plasma renin and norepinephrine concentration, which reflect the activity of the renin-ngiotensin and sympathetic nervous systems respectively, were also measured. RESULTS: The resistive index in the middle cerebral artery were significantly higher in cirrhotic patients with ascites (0.58 +/- 0.04, mean +/- standard deviation) than in cirrhotic patients without ascites (0.53 +/- 0.02, p<0.01) and in control subjects (0.50 +/- 0.05, p<0.01). The resistive index in the middle cerebral artery showed direct correlation with renal resistive index (r = 0.52, p<0.01), plasma renin activity (r = 0.44, p<0.01) and norepinephrine (r = 0.33, p<0.05). The resistive index in the middle cerebral artery showed an inverse correlation with mean arterial pressure (r = -.59, p<0.01). CONCLUSION: The results suggest that in patients of cirrhosis with ascites, independent of the amount of ascites, there is a cerebral vasoconstriction which is related with the arterial hypotension and the overactivity of vasoconstrictor systems.
Arterial Pressure ; Arteries ; Ascites* ; Cardiac Output, High ; Fibrosis ; Hemodynamics ; Humans ; Hypertension, Portal ; Hypotension ; Liver Cirrhosis* ; Liver* ; Middle Cerebral Artery ; Norepinephrine ; Plasma ; Portal Pressure ; Renal Circulation ; Renin ; Sympathetic Nervous System ; Ultrasonography, Doppler ; Vascular Resistance ; Vasoconstriction ; Vasodilation

No abstract available.
Baculoviridae* ; Glycoproteins* ; Insects*

BACKGROUND AND OBJECTIVES: Many people have been concerned about voice change after laser assisted uvulopalatoplasty (LAUP). A number of studies reported acoustic changes after uvulopalatopharyngoplasty (UPPP) and LAUP. However, there have not been any reports on the association between anatomic change and acoustic results after LAUP. The purpose of this study is to analyze changes in the voice and changes in the vocal tract after LAUP and to evaluate whether the anatomical changes of vocal tract have an effect on the voice change or not. SUBJECTS AND METHOD: By using CSL, we analyzed fourteen LAUP cases on the formant frequencies of six vowels (/a/, /i/, /u/, /=, /o/, /e/) and four nasal consonants (/hana/, /eomma/, /eoungga/, /chiken/). By using MR image, we analyzed changes in the vocal tract eight weeks after LAUP with preoperative findings in three cases. RESULTS: In acoustic analysis, the second formant frequencies of /u/ and /= phonation were significantly reduced postoperatively compared to those of preoperative status. In imaging study of /u/ and /=, the uvula and soft palate were contracted, so coupling was occurred between nasal cavity and oropharynx in /u/ and /= phonation and the tongue was shifted toward posterior pharyngeal wall to compensate coupling. CONCLUSION: LAUP reduced the second formant of /u/ and /=, which did not result in serious voice changes.
Acoustics* ; Nasal Cavity ; Oropharynx ; Palate, Soft ; Phonation ; Tongue ; Uvula ; Voice*

Doxorubicin has characteristic chemomyectomy effect of the eyelid without disturbing other eyelid structures, but the major side effect of doxorubicin is the potential for eyelid skin injury as a result of the drug's toxicity in both animal and clinical studies. Verapamil may be used to reduce the dose of doxorubicin and the number of injections that would amplify the toxic effects of doxorubicin. This study was performed to determine whether there is an increase in the toxic effect of the doxorubicin as a result of verapamil pretreatment of the muscle. After 0.5mg, 1.0mg, and 2.0mg doxorubicin was injected in lower eyelids of each group, and equal dose of doxorubicin was injected fo11owing 1.0mg of verapamil injection in lower eyelid of each group, muscle cell loss were measured by light microscopy and side effect was observed. In verapamil and doxorubicin injection group, there was significant differences in the amount of preseptal muscle and even in the pretarsal muscle than the doxorubicin injection group in all doxorubicin doses. Verapamil, injected with a range of doses of doxorubicin, caused suhstantia11y increased muscle loss in the eyelid, compared with injection of doxorubicin alone. Skin ulceration, entropion or ectropion were not visible. Clinically, verapamil cotreatment might be useful to decrease the dose of doxorubicin injected and/or the total number of injections.
Animals ; Doxorubicin* ; Ectropion ; Entropion ; Eyelids ; Microscopy ; Muscle Cells ; Skin ; Skin Ulcer ; Verapamil*

BACKGROUND AND OBJECTIVES: High-resolution computed tomography provides an excellent method for examination of the middle ear, inner ear anatomy and pathologic changes in the temporal bones. The purpose of this study was to get various measurements of the external auditory canal and temporal bones, and to compare the changes with age and sex. MATERIALS AND METHOD: The various measurements by CT of the external ear canal of normal 50 ears were done. RESULTS: The obtained results were as follows: 1) In both male and female group, the distance from the medial end of EAC to bony-cartilage junction, the distance from medial end of EAC to the lateral end of cartilage, the distance from the superior wall to the inferior wall (coronal EAC isthmus) and the distance from anterior wall to the posterior wall (axial EAC isthmus) increased significantly with age (p<0.05). 2) Compared with the female group, the male group had longer distance from the posterior tympanic plate to the sinus tympani with age (p<0.05). 3) Compared with male group, the female group had longer distance from the posterior EAC wall to the Sigmoid sinus and from the EAC superior wall to the tegmen tympani with age (p<0.05). 4) In both male and female groups, the anterior and inferior angles increased and the posterior and superior angles decreased with age (p<0.05). CONCLUSION: Computed tomographic evaluation can give us to information to operate external ear, middle ear, and inner ear surgery.
Cartilage ; Colon, Sigmoid ; Ear ; Ear Canal* ; Ear, External ; Ear, Inner ; Ear, Middle ; Female ; Humans ; Male ; Temporal Bone*

The Japanese encephalitis virus (JEV) is one of causative agents of reproductive failure in pregnant sows. An indirect enzyme-linked immunosorbent assay (I-ELISA) was examined for its potential use in the rapid monitoring of the JEV, and the results were compared with those from the hemagglutination inhibition (HI) and serum neutralization (SN) tests. The comparative analysis showed that the results of I-ELISA showed a significant correlation with the conventional HI (r = 0.867) and SN tests (r = 0.804), respectively. When the I-ELISA results were compared with the traditional diagnostic assays, the sensitivity of the I-ELISA was 94.3% with the HI test and 93.7% with the SN test, respectively. The specificity was found to be 81.4% and 80.0% with the HI and SN tests, respectively. To determine the applicability of I-ELISA in the field, the serum samples from 720 pigs were collected from 4 regions in Korea between July and August 2004. The results indicated that 21.7% of screened pigs were seropositive for the JEV. The seropositive rates of JEV in the 4 provinces were 12.6% in Gyeonggi, 45.0% in Gyeongnam, 16.7% in Jeonbuk, and 12.2% in Jeju. The I-ELISA methodology developed in this study was shown to have considerable sensitivity and specificity through a comparison with HI and the SN tests. Therefore, it might be one of convenient methods for screening a large number of samples in various fields.
Animals ; Antibodies, Viral/blood ; Antigens, Viral/immunology ; Encephalitis Virus, Japanese/immunology/*isolation&purification ; Encephalitis, Japanese/blood/immunology/*veterinary/virology ; Enzyme-Linked Immunosorbent Assay/methods/*veterinary ; Female ; Hemagglutination Inhibition Tests/veterinary ; Korea ; Neutralization Tests/veterinary ; Swine ; Swine Diseases/blood/immunology/*virology

One step TaqMan reverse transcription polymerase chain reaction (RT-PCR) using TaqMan probe was developed for detection of Japanese encephalitis virus (JEV). Real-time RT-PCR was optimized to quantify JEV using the detection system (Rotor Gene 2000 detector) and dual-labeled fluorogenic probes. The gene specific labeled fluorogenic probe for the 3' non-translated region (3' NTR) was used to detect JEV. When the specificity of the assay using specific JEV primers was evaluated by testing three different JEV strains, other swine viruses and bovine viral diarrhea virus, no cross-reactions were detected with non-JE reference viruses. A single tube TaqMan assay was shown to be 10-fold more sensitive than the conventional two-step RT-PCR method. Detection limits of two step and real-time RT-PCR for JEV were 112 TCID50 /ml and 11.2 TCID50 /ml, respectively. Quantification of JEV was accomplished by a standard curve plotting cycle threshold values (Ct ) versus infectivity titer. Real-time RT-PCR assay using single tube method could be used as a sensitive diagnostic test, and supplied the results in real time for detection and quantification of JEV. We could detect JEV RNA genome in plasma samples of pigs inoculated with KV1899 strain at 2 days post inoculation, but couldn't in 41 fetus samples. This assay was sensitive, specific, rapid and quantitative for the detection of JEV from laboratory and field samples.
Animals ; DNA Primers/chemistry ; DNA Probes/chemistry ; Encephalitis Virus, Japanese/genetics/*isolation&purification ; Encephalitis, Japanese/diagnosis/*veterinary/virology ; RNA, Viral/analysis ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction/methods/*veterinary ; Sensitivity and Specificity ; Swine ; Swine Diseases/*diagnosis/virology ; *Taq Polymerase

Genes encoding for the premembrane and envelope (prME), envelope (E) and nonstructural protein (NS1) of Japanese encephalitis virus (JEV) were cloned. Each protein was expressed in baculovirus expression system. Of the three proteins expressed in baculovirus system, only prME had hemagglutination activity. The prME (72 and 54 kDa), E (54 kDa) and NS1 (46 kDa) proteins could be detected by Western blotting in the recombinant virus infected cells. Immunogenicity of the recombinant proteins obtained from infected Spodoptera frugiperda (Sf-9) cells was examined in mice. The 3 week-old ICR mice immunized intraperitoneally with three recombinant proteins three times were challenged with a lethal JEV. A survival rate was increased from about 7.7% in unimmunized mice to 92.3% in E + prME and only E groups. The complete protection was shown in prME and live vaccine inoculated groups, respectively. We also measured neutralizing antibody and three immunoglobulin subtypes of IgG1, IgG2a and IgG2b in the sera of mice before and after challenge. Titers of IgG1 antibodies were approximately two to three times higher than that of IgG2b antibodies in all the immunized groups as compared to the control group. However, IgG2a antibody level somewhat increased after challenge, indicating T-helper type 1 (Th1) cell response. The results of this study can provide useful information for developing efficacious subunit vaccine against JEV.
Animals ; Antibodies, Viral/blood ; Baculoviridae/genetics ; Blotting, Western ; Cloning, Molecular ; Encephalitis Virus, Japanese/genetics/*immunology ; Encephalitis, Japanese/*immunology/prevention&control ; Female ; Immunization ; Immunoglobulin Isotypes/blood ; Japanese Encephalitis Vaccines/*immunology/standards ; Mice ; Mice, Inbred ICR ; Microscopy, Fluorescence ; Plasmids ; Recombinant Proteins/genetics/immunology ; Viral Envelope Proteins/genetics/*immunology ; Viral Matrix Proteins/genetics/*immunology ; Viral Nonstructural Proteins/genetics/*immunology

We have determined the complete nucleotide and deduced amino acid sequences of the Japanese encephalitis virus (JEV) strain KV1899, isolated from a fattening pig in Korea. In comparison with 22 fully sequenced JEV genomes currently available, we found that the 10,963-nucleotide RNA genome of KV1899 has a 13-nucelotide deletion in the 3' non-translated variable region and 53 unique nucleotide sequences including 3' non-translated region (NTR). Its single open reading frame has a total of 28 amino acid substitutions. Comparison of the KV1899 genomic sequence with those of the 21 fully sequenced JEV strains in published databases showed nucleotide homology ranging from 97.4% (Ishikawa strain) to 87.0% (CH2195 strain). Amino acid homology with KV1899 strain ranged from 96.4% (K94P05) to 91.0% (GP78). The KV1899 showed the highest nucleotide homology with Ishikawa strain and the highest amino acid homology with K94P05. We performed an extensive E gene based phylogenetic analysis on a selection of 41 JEV isolates available from the GenBank. Compared with Anyang strain, isolated from a pig in 1969, that is current live vaccine strain for swine in Korea, the homology of nucleotide sequence in envelope gene was only 87.1%. The prM gene of the isolate was closely related with those of Ishikawa and K94P05 strains, which were grouped into genotype I of JEV.
3' Untranslated Regions/chemistry/genetics ; Amino Acid Sequence ; Animals ; Base Sequence ; Culicidae/virology ; Encephalitis Virus, Japanese/*genetics ; Encephalitis, Japanese/*veterinary/virology ; *Genome, Viral ; Humans ; Korea ; Membrane Glycoproteins/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Viral/chemistry/genetics ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Alignment ; Swine ; Swine Diseases/*virology ; Viral Envelope Proteins/chemistry/genetics