Resveratrol protects SH-SY5Y neuroblastoma cells from apoptosis induced by dopamine

Dopamine (DA) is an oxidant that may contribute to the degeneration of dopaminergic neurons. The present study demonstrates that DA-induced cytotoxicity in human-derived neurotypic cells, SH-SY5Y, is prevented by resveratrol, one of the major antioxidative constituents found in the skin of grapes. SH-SY5Y cells, a neuroblastoma cell line, treated with DA at 300 and 500 micrometer for 24 h underwent apoptotic death as determined by characteristic morphological features, including nuclear condensation, and loss of mitochondrial membrane potential (MMP). Flow cytometric analysis using Annexin V showed that DA can induce significant and severe apoptosis. Exposure to resveratrol (5 micrometer) for 1 h prior to the DA treatment attenuated DA-induced cytotoxicity, and rescued the loss of MMP. To investigate the apoptotic signaling pathways relevant to the restoration of DA-induced apoptosis by resveratrol, we carried out quantitative analysis of Bcl-2, caspase-3, and cleaved poly ADP-ribose polymerase (PARP) by immunoblot analysis. Resveratrol pretreatment led to a decrease in cleavage of PARP, an increase in the Bcl-2 protein, and activation of caspase-3. These results suggest that DA may be a potential oxidant of neuronal cells at biologically relevant concentrations. Resveratrol may protect SH-SY5Y cells against this cytotoxicity, reducing intracellular oxidative stress through canonical signal pathways of apoptosis and may be of biological importance in the prevention of a dopaminergic neurodegenerative disorder such as Parkinson disease.

Nucleotide Sequence and Phylogenetic Analysis of M and L Segment of Soochong Virus

Hantaan virus (HTNV) and Seoul virus (SEOV) have mainly been known as the cause of hemorrhagic fever with renal syndrome (HFRS) in Korea since HTNV has been isolated from Korean field mouse, Apodemus agrarius in 1976 and SEOV has been isolated from Rattus norvegicus in 1980. Soochong virus-1, -2, -3, -4 (SOOVs) were isolated from lung tissues of four Apodemus peninsulae captured on August 1997 at Mt. Gyebang in Hongcheon-gun, Mt. Gachil, Inje-gun, Gangwon Province, and in September 1998 at Mt. Deogyu, Muju-gun, Jeollabuk Province. Apodemus peninsulae is the second-most dominant field rodent species found throughout Korea. To determine phylogenetic analysis of SOOVs, we entirely identified nucleotide sequences of M and L segments. The length of M segment was 3,615 bp and L segment was 6,533 bp. SOOVs were diverged from HTNV by 22.7~23.3% and SEOV by 36.3~37.2%, in M segment. In addition, L segment was diverged from HTNV by 21.8~22.0% and SEOV by 30.3~30.5%. SOOVs sequence compared with Amur virus (AMRV) in M segment showed that SOOVs were different with AMRV about 14.6~16.2% in nucleotide sequences. Neighbor-joining phylogenetic analysis based on entire sequences of the M and L segment indicated that the SOOVs sequences present a separate lineage with HTNV, SEOV and AMRV. SOOVs constituted an individual cluster on the phylogenetic tree and they composed a phylogenic lineage separately. According to these data, SOOVs could be classified as a new hantavirus.

Retraction: Nucleotide Sequence and Phylogenetic Analysis of M and L Segment of Soochong Virus

No abstract available.

In vivo characterization of the integrin beta3 as a receptor for Hantaan virus cellular entry

Binding of viruses to cell surface molecules is an essential step in viral infection. In vitro studies suggested that the alpha v beta3 integrin receptor is the epithelial cell receptor for Hantaan virus (HTNV). Whether beta3 is in vivo the only or central cellular receptor for HTNV infection is not known. To investigate the role of beta3 integrin for cellular entry of HTNV, we established an HTNV infection model in newborn murine pups. Infected pups died at an average age of 14.2 +/- 1.1 days with high levels of viral antigen detected in their brain, lung, and kidney. Pre-injection of blocking monoclonal antibodies (mAb) specific for either beta3 or av prolonged survival significantly to a maximal average survival of 19.7 +/- 1.5 days (P<0.01) and 18.4 +/- 0.9 days (P<0.01), respectively. XT-199, a chemical blocker of the alpha v beta3 receptor also prolonged survival to 19.5 +/- 1.3 days (P<0.01). In contrast to these receptor blockades, anti-HTNV antibody was not only able to prolong survival, but 20% of infected pups achieved long-term survival. An anti-murine beta1 antibody comparatively prolonged survival (19.0 +/- 1.2 days), suggesting that HTNV infection is partly mediated through integrin beta1 receptors as well as through beta3 receptors in vivo. Our data demonstrate that the beta3 receptor is important for HTNV infection in vivo, but also suggest that HTNV may utilize additional receptors beyond beta3 for cellular entry within an organism.

Serological Characterization of Soochong and Muju Virus as New Serotype of Hantavirus

Apodemus agrarius, which accounts for three-fourths of the wild rodents, mainly inhabits in cultivated fields of Korea. Apodemus peninsulae and Eothenomys regulus are the second and third dominant species, respectively. Soochong virus (SOOV) from A. peninsulae and Puumala-related Muju virus (MUJV) from E. regulus were isolated in 1997 and 1998 in Korea, respectively. But serological characterizations of SOOV and MUJV were not identified clearly. Thus, in order to determine the serotypic classification, simultaneous cross-indirect immunofluorescent antibody (IFA) assay and cross-plaque reduction neutralization (PRN) test against four different hantaviruses were conducted with sera from 17 A. agrarius, 19 A. peninsulae, and 8 E. regulus strains. IFA titers of sera from A. agrarius and A. peninsulae were the highest to Hantaan virus (HTNV) and SOOV, respectively. However, most sera showed similar IFA titers to Seoul virus (SEOV). Therefore it was difficult to do serotyping using the sera from A. agrarius and A. peninsulae by IFA. In case of sera of E. regulus, IFA titers to Puumala virus (PUUV) were higher than HTNV, SOOV and SEOV. Cross-PRN result of A. agrarius to HTNV, SOOV, SEOV and PUUV was 6,890, 5,120, 110 and 30, respectively. In case of A. peninsulae, the mean PRN titer was the highest to SOOV (1:6,820) and those to HTNV, SEOV and PUUV were 1,580, 100 and 30, respectively. The mean PRN titers of E. regulus to HTNV, SOOV, SEOV and PUUV were 70, 10, 80 and 640. SOOV and MUJV could be distinguished from HTNV and SEOV by cross-PRNT. These results demonstrate that SOOV and MUJV could be classified as new serotype of hantavirus.

Characterization of E. coli-induced Genes in Bombyx mori Fatbody using Expressed Sequence Tags Analysis

To accelerate the molecular analysis of specifically induced antibacterial peptide against pathogen (E. coli), cDNA library prepared from the larvae fatbody of Bombyx mori was examined by the expressed sequence tag (EST) analysis. In a total of 722 clones, 653 clones were unique genes. Of 653 unique genes, 43.2% (282/653 ESTs) was identified as characterized genes, 38.1% (249/653 ESTs) as uncharacterized genes, and 18.7% (122/653 ESTs) as novel ESTs. According to the functional categorization of the characterized genes, 36.2% (102/282 ESTs) was antibacterial proteins. The highest expressed peptides, 78.4% of all the expressed antibacterial proteins (80/102 ESTs), belonged to the cecropin family. The antibacterial effect of selected clones representing novel ESTs based on a phylogenetic analysis was examined against various bacterial strains. None of the clones showed significant inhibitory effect to the bacteria tested. These results suggested that most of the novel molecules induced by E. coli may not act as immune-induced antibacterial peptides in the fatbody.

Epidemilogical Study on Orientia tsutsugamushi Infection of Wild Rodents in Korea

This study was carried out to determine the seropositivity for Orientia tsutsugamushi infection among wild rodents captured in farms and mountains of Korea. A total 965 wild rodents of 7 species and 48 Crocidura laciura were captured in farms and mountains of 6 province during 1997~2000 in Korea. Serological evidence for O. tsutsugamushi infection was obtained using O. tsutsugamushi antigens by indirect immunofluorescent antibody technique (IFA). Four hundred eighty four Apodemus agrariusus, 246 Apodemus peninsulae, 179 Eothenomys regulus, 44 Microtus fortis, 6 Micromys minutus, 5 Mus musculus, and 1 Cricetula triton were captured in Gyeonggi, Gangwon, Chungnam, Gyeongbuk, Gyeongnam, and Jeonbuk province. Among them, 59 (12.2%) A. agrariusus, 22 (8.9%) A. peninsulae, 12 (6.7%) E. regulus, 5 (11.4%) M. fortis and 1 (16.7%) M. minutus were IF antibody positive against O. tsutsugamushi Gilliam or Karp strain.

Genomic organization and expression of parkin in Drosophila melanogaster

We report here the isolation, characterization on genomic structure and expression of the D. melanogaster homolog of human parkin. The 2,122 bp parkin gene sequence contains six exons that form a 1,449 bp transcript encoding a protein of 482 amino acids. 151 bp of 5' and 112 bp of 3' untranslated regions were identified by a combination of 5'-RACE/primer extension and 3'-RACE, respectively. The 5' UTR contains three transcription initiation sites. Neither a classical TATA nor a CAAT box was found in the putative promoter sequence. However, binding sites for AhR-Arnt, AP4, NF1 and GATA transcription factors were identified. Transient transfection analysis of the 5' UTR confirmed its promoter activity in HEK 293 cells and SH-SY5Y neuronal cells using a dual luciferase reporting system. The amino acid sequence of D. melanogaster Parkin exhibits 42%, 43% and 43% identity to that of human, mouse and rat, respectively, representing a 54 kDa protein band via western blot analysis. It shows a high degree of conservation in the Ubiquitin-like domain at the N-terminus (34%), the In-Between RING finger domains (IBR, 65-69%), and the RING finger domains at the C-terminus (56-57%). The expression pattern of D. melanogaster parkin varies during the developmental stages, with the highest expression in the adult stage as measured by competitive RT-PCR. From immunostainings of the embryo, D. melanogaster parkin was expressed slightly higher in the central nervous system (brain and nerve cord) during the late embryonic stage.

Seroepidemiologic Analysis of Acute Febrile Illness in Korea during 1997~1998

Hemorrhagic fever with renal syndrome (HFRS), scrub typhus, murine typhus and leptospirosis have been the principal acute febrile diseases in Korea for many years. To evaluate the seroepidemiologic patterns of the acute febrile illness, sera collected from 4,503 patients in 1997~1998 were examined for antibodies against Hantaan virus, Orientia tsutsugamushi and Rickettsia typhi by indirect immunofluorescent antibody technique (IFA) and macroscopic agglutination test for Leptospira interogans. Seropositive cases for Orientia tsutsugamushi, Rickettsia typhi, Leptospira interogans and Hantaan virus were 261 (12.4%), 242 (11.5%), 11 (0.5%), and 250 (11.9%) in 1997, and 415 (17.3%), 273 (11.4%), 16 (0.7%), and 357 (14.9%) in 1998, respectively. Male was affected more frequently by HFRS and leptospirosis while scrub typhus was more prevalent in female. Old age group was more susceptible to the acute febrile diseases. Most positive cases were occurred during October and November for scrub typhus, and during November and December for HFRS. These results showed similar patterns with previous epidemiological data obtained during recent several years, except the single scrub typhus epidemic in 1998, and implied that no significant changes occurred in ecologic system for acute febrile diseases in Korea.

Seroprevalence and Phylogenetic Analysis of Orientia tsutsugamushi from Apodemus peninsulae in Korea

To investigate the seroprevalence of the Orientia tsutsugamushi infection of Apodemus peninsulae and genomic variations in O. tsutsugamushi isolates, 246 A. peninsulae were trapped in 14 mountainous areas approximately 500 meter above sea level in Korea during the period of 1997 and 2000. Seropositive rate of O. tsutsugamushi among A. peninsulae was 31.8% in Kyunggi, 8.2% in Chunbuk and 7.1% in Kangwon provinces by microimmunofluorescent test. The 56 kDa protein gene was amplified by PCR in the spleens of seropositive A. peninsulae. Two amplicons from seropositive A. peninsulae were sequenced and their phylogeny was analysed on the basis of sequence homology. The 56 kDa genes of A. peninsulae 98-12 strain and A. peninsulae 98-16 strain showed 98.7% nucleotide homology and 96.6% amino acid similarity. A. peninsulae 98-12 and A. peninsulae 98-16 strain were related to Kuroki, Boryong and Karp strains showing 93.3~92.2%, and 87.1~84.6% homologies in nucleotide and amino acids levels, respectively. In the phylogenetic analysis, A. peninsulae 98-12 and A. peninsulae 98-16 strain formed a distinct group with Boryong, Kuroki and Nishino strains and were clearly distinguished from other genetic groups. The results suggest that A. peninsulae might be an important reservoir of O. tsutsugamushi in Korea.

Isolation of Apodemus peninsulae-borne Hantavirus and Cimparison of Molecular Biological Characteristics

Two distinct hantaviruses have been isolated from Apodemus agrarius in 1976 and Rattus norvegicus in 1980 in Korea. Since our serosurveys conducted in 1994, a genetically distinct hantavirus from Apodemus peninsulae has been investigated. To isolate hantavirus from A. peninsulae captured in Korea, the lung homogenate of seropositive A. peninsulae inoculted Vero E6 cells. Viral antigen was detected in a progressively higher percentage of cells with subsequent passage after 80 days postinoculation. The new isolate from seropositive Apodemus peninsulae was designated Suchong virus after Suchong valley located in northeastern region of South Korea. Comparing with hantaan virus 76-118 strain, Suchong virus-1, 2, 3 and 4 showed the similarity of 71.0~91.8% at nucleotide and 90.9~94.8% at amino acid sequences in 231 nucleotides region of M segment, and the similarity of 75.1~81.0% at nucleotide and 97.5~100% at amino acid sequences in 237 nucleotides of S segment.

Sequence Analysis of Rubella Viruses Isolated in Korean and Expression of Rubella Specific Gene

During the recent epidemic period (1995~1996), seven strains of rubella virus were isolated in Korea. To analyze phylogenetic relationship between seven Korean strains and rubella virus strains from other different geographical areas, structural genes (E1, E2 and C) of Korean strains were enzymatically amplified and automatically sequenced. The sequence similarities of the E1, E2 and C genes of the cosmopolitan types were 95.8~98.1%, 92.6~99.2% and 96.4~99.3% based on 1,441, 122 and 139 nucleotides and 96.9~98.5%, 90~100% and 97.8~100% based on 480, 40 and 46 amino acids compared to the sequences of strain RA27/3, respectively. In contrast, the sequence similarities of the E1, E2 and C genes of the Asian types were 91.5~92.1%, 83.6~88.5% and 91.4% based on nucleotides ad 96.9~97.7%, 85.5% and 97.8% based on amino acids compared to the sequences of strain RA27/3. respectively. However, immunodominent epitopes of the E1 gene of the cosmopolitan and Asian types were well conserved, and the growth patterns in cell culture and immunofluorescent antibody titers in cross-reaction test showed no differences between two different types. In phylogenetic analysis based on nucleotide sequences of each gene regions, the comopolitan and Asian types formed tow distinct phylogenetic lineages. These data showed two distinct genotypes of rubella viruses cocirculated in Korea, but no significant differences in the antigenicity of two different rubella virus strains were found.

Seroepidemiological Study on Hantavirus Infection of Wild Fodents Captured in the Moutainous Areas of Korea

Hantaan virus is widely distributed among rodent populations in Korea. Two antigenically distinct hantaviruses were isolated from Apodemus agrarius in 1976 and Rattus norvegicus in 1980 in Korea. This study was designed to investigate the serological evidence of hantavirus infection among indegenous wild rodents, which were captured in 11 mountains located in Kyunggi, Kangwon, Chungnan, Chunbug and Kyungnam province of South Korea. A total 252 wild rodents of 3 species were trapped from Myungsung Mt., Chumbong Mt., Kali Mt., Hansuk Mt., Chachil peak, Kyebang Mt., Odae Mt., Kyerong Mt., Kaya Mt., Togju Mt. and Chiri Mt. in 1997. Serologic test for hantavirus infection was performed using hantavirus antigens by indirect immunofluorescent antibody technique. Among 122 Apodemus agrarius, 88 Apodemus peninsulae and 42 Eothenomys regulus; 18 A. agrarius (14.8%), 12 A. peninsulae (13.6%) and 4 E. regulus (9.5%) were immunofluorescent antibody (IFA) positive against hantaan virus. IFA titers 3 Eothenomys regulus sera were higher against puumalavirus than hantaan virus. These data imply that above three species of rodent might be natural reservoirs of hantaviruses in Korea.

Retraction: Asp 280 residue is important in the activity of the Escherichia coli leader peptidase

No abstract available.

Asp 280 residue is important in the activity of the Escherichia coli leader peptidase

Leader peptidase is a novel serine protease in Escherichia coli, which catalyzes the cleavage of amino-terminal signal sequences from exported proteins. It is an integral membrane protein containing two transmembrane segments with its carboxy-terminal catalytic domain residing in the periplasmic space. Recently, the x-ray crystal structure of signal peptidase-inhibitor complex showed that Asp 280, a highly conserved consensus sequence of E. coli leader peptidase is the closest charged residue in the vicinity of two catalytic dyad, Ser 90 and Lys 145, and it is likely held in place by a salt bridge to Arg 282. Possible roles of Asp 280 and Arg 282 in the structure-catalytic function relationship were investigated by the site-directed mutagenesis of Asp 280 substituted with alanine, glutamic acid, glycine, or asparagine and of Arg 282 with methionine. All of mutants purified with nickel affinity chromatography were inactive using in vitro assay. It is surprising to find complete lose of activity by an extension of one carbon units in the mutant where Asp 280 is substituted with glutamic acid. These results suggest that Asp 280 and Arg 282 are in a sequence which constitutes catalytic crevice of leader peptidase and are essential for maintaining the conformation of catalytic pocket.

Sequence and Phylogenetic Analyses of Rubella Viruses Isolated from 1995 through 1996 in Korea / 감염

BACKGROUND: The age group of Rubella virus (RV) infection in most industrialized nations located in temperate climates, has in large part shifted from children to young adults since introducing of the RV vaccine in late 1960's. Interestingly, there were rubella outbreaks from 1995 through 1996 in Korea, and middle and high school students were mostly affected during that time. Although continued cycling of the rubella epidemics, there were no reports about the isolation or genetic information of rubella viruses circulated in Korea. METHOD: To isolate RV circulated in Korea, and determine the phylogenetic relationship between RV strains in Korea and RV isolates from other geographic regions including vaccine strains, we inoculated nasopharyngeal secretion samples from clinically diagnosed rubella patients to Vero E6 cells, and sequenced corresponding region of the 5' E1 encoding genomic regions of RV isolates. RESULT: Seven RV strains isolated from Korea showed 93.6 to 97.8% and 98.3 to 100% sequence homologies in nucleotide and amino acid levels, respectively, compared to RA27/3 vaccine strain. Phylogenetic tree based on 359bp of RV indicated that at least two different groups of RV circulated in Korea during 1995-1996 epidemics. CONCLUSION: Our data suggested that mutant RV strains were possibly not the cause of recent rubella epidemics in Korea.

Study on Hantavirus Infection of Wild Rodents in Korea / 감염

BACKGROUND: Hantavirus are widely distributed in rodents populations even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Multiple species of Murid and Arvicolid rodents serve as the natural reservoirs of hantaviruses. Serologic diagnosis using hantaviral antigens indicates that hantaviruses are widely distributed in wild rodents. This study was designed to find the distribution of hantaviruses among wild rodents and small mammals in Korea, 1995-1996. METHODS: Rodents were trapped alive in selected areas. A total of 551 wild rodents from 7 species and 97 small mammals from 4 species were captured in Korea. Serologic evidence for hantavirus infection were tested using five hantavirus antigens by indirect immunofluorescent antibody technique (IFA). Among 162 Apodemus agrarius, 23 Apodemus peninsulae, 8 Clethrionomys regulus, 6 Microtus fortis, 1 Mus musculus, 283 Tamias sibiricus, 68 Sciurus vulgaris, 14 Crocidura laciura, 80 Lepus sinensis, 2 Capereolus capereolus and 1 Nyctereutes procyonoides. RESULTS: 29 A. agrarius, 2 A. peninsulae, 1 C. laciura, 2 C. regulus, 27 T. sibiricus and 7 S. vulgaris were seropositive against Hantaan virus and 7 L. sinensis were IF antibody positive against Seoul virus. Some of Tamias sibiricus were only seropositive against Puumala virus or prospect hill virus. CONCLUSION: This data suggests that new serotypes of hantavirus might distribute among rodents in Korea.

Endoscopic Removal of Esophageal Foreign Body Complicated with Esophageal Ulcer: Case report / 대한소화기내시경학회지

Any foreign body in the esohagus means an acute danger because of the impending perforation followed by mediastinitis, the impending erosion of bigarteries, and impending aspiration in the care of esorhageal occlusion. Nowadays, foreign body can reliably be removed hy endoscopy, and do not perforate the wall, not changed their form. Complication at the endoscopic extraction of foreign body, that requre surgical intervention are very rare. We presented a case of esophageal foreign body complicated with esophageal ulcer in a 83 years old male.

Multiple Primary Malignant Neoplasms: A case of double primary malignant cancer / 대한소화기내시경학회지

This is a case report of double primary malignant cancer occurred aynchronously in the stomach and lymphoid tissue, We report this case with review of literatures about the criteria, age distribution, predisposing factor, inidence, immunity and susceptibility of the primary malignant neoplasms. This case was a 59-year-old man who had Hodgkins disease and tubular adenocarcinoma, of the stomach. The diagnoais was verified histologically, Although multiple primary neoplaas are rare, the possibility of that must be conaidered seriously, And its hereditary predisposition and other predisposing factor muat be researched with enthuaiasm.