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1.
Journal of Korean Medical Science ; : 1190-1196, 2016.
Article in English | WPRIM | ID: wpr-69712

ABSTRACT

Diagnosis of scrub typhus is challenging due to its more than twenty serotypes and the similar clinical symptoms with other acute febrile illnesses including leptospirosis, murine typhus and hemorrhagic fever with renal syndrome. Accuracy and rapidity of a diagnostic test to Orientia tsutsugamushi is an important step to diagnose this disease. To discriminate scrub typhus from other diseases, the improved ImmuneMed Scrub Typhus Rapid Diagnostic Test (RDT) was evaluated in Korea and Sri Lanka. The sensitivity at the base of each IgM and IgG indirect immunofluorescent assay (IFA) in Korean patients was 98.6% and 97.1%, and the specificity was 98.2% and 97.7% respectively. The sensitivity and specificity for retrospective diagnosis at the base of IFA in Sri Lanka was 92.1% and 96.1%. ImmuneMed RDT was not reactive to any serum from seventeen diseases including hemorrhagic fever with renal syndrome (n = 48), leptospirosis (n = 23), and murine typhus (n = 48). ImmuneMed RDT shows superior sensitivity (98.6% and 97.1%) compared with SD Bioline RDT (84.4% at IgM and 83.3% at IgG) in Korea. The retrospective diagnosis of ImmuneMed RDT exhibits 94.0% identity with enzyme-linked Immunosorbent assay (ELISA) using South India patient serum samples. These results suggest that this RDT can replace other diagnostic tests and is applicable for global diagnosis of scrub typhus. This rapid and accurate diagnosis will be beneficial for diagnosing and managing scrub typhus.


Subject(s)
Humans , Antibodies, Bacterial/blood , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique, Indirect , Immunoglobulin G/blood , Immunoglobulin M/blood , Orientia tsutsugamushi/immunology , Reagent Kits, Diagnostic , Retrospective Studies , Scrub Typhus/diagnosis , Sensitivity and Specificity
2.
Journal of Korean Medical Science ; : 183-189, 2016.
Article in English | WPRIM | ID: wpr-133739

ABSTRACT

Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.


Subject(s)
Female , Humans , Male , Antigens, Bacterial/immunology , Argentina , Bulgaria , Enzyme-Linked Immunosorbent Assay , Leptospira/isolation & purification , Leptospira interrogans/isolation & purification , Leptospirosis/diagnosis , Polysaccharides/immunology , Reagent Kits, Diagnostic/standards , Republic of Korea , Sensitivity and Specificity
3.
Journal of Korean Medical Science ; : 183-189, 2016.
Article in English | WPRIM | ID: wpr-133738

ABSTRACT

Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.


Subject(s)
Female , Humans , Male , Antigens, Bacterial/immunology , Argentina , Bulgaria , Enzyme-Linked Immunosorbent Assay , Leptospira/isolation & purification , Leptospira interrogans/isolation & purification , Leptospirosis/diagnosis , Polysaccharides/immunology , Reagent Kits, Diagnostic/standards , Republic of Korea , Sensitivity and Specificity
4.
Journal of Bacteriology and Virology ; : 23-36, 2014.
Article in English | WPRIM | ID: wpr-70413

ABSTRACT

Human immunodeficiency virus type 1 (HIV-1)-infected cells respond to the infection with different outcomes depending on their cell type. The interplay of cellular and viral proteins is a key player of differences in virus replication and disease progression. Myeloid cells, including monocytes, macrophages, and myeloid dendritic cells (mDCs) play a crucial role in the transmission and pathogenesis of HIV. The viral protein Tat, which is the viral transcriptional activator, modulates the expression of both HIV and cellular genes in these myeloid cells. This review will focus on recent advances on the interplay between HIV and myeloid cells and will discuss how this interaction may contribute to HIV pathogenesis. A better understanding of the pathogenesis of HIV disease will provide us with the scientific rationale for novel approaches to prevention.


Subject(s)
Humans , Dendritic Cells , Disease Progression , HIV , HIV-1 , Macrophages , Monocytes , Myeloid Cells , Viral Proteins , Virus Replication
5.
Journal of Bacteriology and Virology ; : 59-66, 2014.
Article in Korean | WPRIM | ID: wpr-112745

ABSTRACT

In this study, we selected only serologically identified 15 Leptosira interrogans isolates in the past and analyzed and identified them by using molecular method. The partial 16S rDNA and LipL32 genes were amplified from the bacteria by polymerase chain reaction (PCR) and sequenced. Sizes of the PCR products were 529 bp and 819 bp respectively and analysis of the nucleotide sequence of 16S rDNA and LipL32 genes showed that 14 out the 15 Leptospira showed 99.4~100% and 99.2~99.9% similarity respectively to those of L. interrogans lai and one isolate named HS-7 showed 100% and 100% similarity to L. interrogans canicola. The phylogenetic tree based on the 16S rDNA and LipL32 genes obtained the study revealed that 14 of the Leptospira composed a cluster distinct to that of L. interrogans lai and HS-7 composed to L. interrogans canicola.


Subject(s)
Bacteria , Base Sequence , DNA, Ribosomal , Leptospira interrogans , Leptospira , Polymerase Chain Reaction
6.
Journal of Korean Medical Science ; : 672-679, 2013.
Article in English | WPRIM | ID: wpr-65460

ABSTRACT

Diagnosis of scrub typhus is difficult because its symptoms are very similar to other acute febrile illnesses, such as leptospirosis, murine typhus, and other viral hemorrhagic fevers. To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We have developed a chimeric recombinant antigen cr56 and two other recombinant antigens, r21 and kr56, from various serotypes of Orientia tsutsugamushi. They were tested for the detection of antibodies against O. tsutsugamushi in the patient's serum samples using enzyme-linked immunosorbent assay (ELISA) and dot-blot analyses. As of conventional immunofluorescence assay (IFA), when the mixture of these three recombinant antigens was used, both sensitivity and specificity of the recombinant antigens were increased up to 98% in IgM and IgG at ELISA and dot blotting. Additionally, both sensitivity and specificity by detection of IgM and IgG antibodies at rapid diagnostic test (RDT), using the mixture of three antigens and gold conjugated antibodies, were 99%. Our results suggest the use of mixture of these recombinant antigen proteins in ELISA or RDT is suitable as a diagnostic test for scrub typhus.


Subject(s)
Humans , Antibodies, Bacterial/blood , Antigens, Bacterial , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique, Indirect , Gold/chemistry , Immunoassay , Immunoglobulin G/blood , Immunoglobulin M/blood , Orientia tsutsugamushi/immunology , Recombinant Proteins/biosynthesis , Scrub Typhus/diagnosis , Sensitivity and Specificity , Serotyping
7.
Journal of Bacteriology and Virology ; : 209-212, 2011.
Article in Korean | WPRIM | ID: wpr-181166

ABSTRACT

Viral disease is traditionally thought to result from an insufficient response of the host to infection, leading to increased replication of viruses and consequently disease. However, the disease is not the simple result of uncontrolled replication of virus. Indeed, the inflammatory response triggered by certain infection is frequently the cause of tissue damage and death. This inflammatory situation can be called as immunopathologic phenomena. By understanding the mechanisms which drive disease, novel therapies may be devised for treatment of patients. I will describe the situations of some viral diseases in which unwanted (excessive, misplaced or altered) inflammation causes immunopathologic phenomena and is responsible for disease induction. In these situations, I will also describe some candidates of antiviral drug which inhibit or modulate the inflammatory response to viral infection.


Subject(s)
Humans , Inflammation , Virus Diseases , Viruses
8.
Infection and Chemotherapy ; : 59-67, 2004.
Article in English | WPRIM | ID: wpr-721919

ABSTRACT

BACKGROUND: Many strains of Leptospira interrogans have been isolated in Korea since 1984. Most isolates were identified as serovar lai by serological methods. The pulsed field gel electrophoresis (PFGE) patterns of Korean isolates have not been investigated currently. METHODS: 29 reference strains and 29 Korean isolates of Leptospira interrogans were characterized by PFGE. Chromosomes were digested by the Not I restriction enzyme and subsequently PFGE was performed in CHEF-DRII (Bio Rad Lab) with 3 pulse times (30 seconds 13 hours, 60 seconds 13 hours, 120 seconds 14 hours) at 150 V (4.5 V/cm). RESULTS: 12 serogroup reference strains and most 17 serovars reference strains in the serogroup Icterohaemoffhagie showed the unique Not I restriction patterns. Most isolates identified serologically as serovar lai showed the same PFGE patterns as the serovar lai reference strain. The strain HM3 and 18R identified serologically as new serovars yeonchon and hongchon respectively showed the same PFGE patterns as serovar lai. The strain AP31, CH88-19 and NR13 that were different from serovar lai by serological methods showed the PFGE patterns indistinguishable from serovar lai reference strain. The strain HY2 that was identified as serovar lai, and the strain 30R that was different from serovar lai serologically showed the PFGE patterns slightly different from serovar lai reference strain. CONCLUSION: The PFGE profile of most Korean isolates Leptospira interrogans serologically identified as serovar lai is identical to the reference strain serovar lai. PFGE analysis thus may be applied to identify serovar of isolates and to investigate the genetic diversity of related serovar.


Subject(s)
DNA , Electrophoresis, Gel, Pulsed-Field , Genetic Variation , Korea , Leptospira interrogans , Leptospira
9.
Infection and Chemotherapy ; : 59-67, 2004.
Article in English | WPRIM | ID: wpr-721414

ABSTRACT

BACKGROUND: Many strains of Leptospira interrogans have been isolated in Korea since 1984. Most isolates were identified as serovar lai by serological methods. The pulsed field gel electrophoresis (PFGE) patterns of Korean isolates have not been investigated currently. METHODS: 29 reference strains and 29 Korean isolates of Leptospira interrogans were characterized by PFGE. Chromosomes were digested by the Not I restriction enzyme and subsequently PFGE was performed in CHEF-DRII (Bio Rad Lab) with 3 pulse times (30 seconds 13 hours, 60 seconds 13 hours, 120 seconds 14 hours) at 150 V (4.5 V/cm). RESULTS: 12 serogroup reference strains and most 17 serovars reference strains in the serogroup Icterohaemoffhagie showed the unique Not I restriction patterns. Most isolates identified serologically as serovar lai showed the same PFGE patterns as the serovar lai reference strain. The strain HM3 and 18R identified serologically as new serovars yeonchon and hongchon respectively showed the same PFGE patterns as serovar lai. The strain AP31, CH88-19 and NR13 that were different from serovar lai by serological methods showed the PFGE patterns indistinguishable from serovar lai reference strain. The strain HY2 that was identified as serovar lai, and the strain 30R that was different from serovar lai serologically showed the PFGE patterns slightly different from serovar lai reference strain. CONCLUSION: The PFGE profile of most Korean isolates Leptospira interrogans serologically identified as serovar lai is identical to the reference strain serovar lai. PFGE analysis thus may be applied to identify serovar of isolates and to investigate the genetic diversity of related serovar.


Subject(s)
DNA , Electrophoresis, Gel, Pulsed-Field , Genetic Variation , Korea , Leptospira interrogans , Leptospira
10.
Asian Journal of Andrology ; (6): 29-34, 2004.
Article in English | WPRIM | ID: wpr-300869

ABSTRACT

<p><b>AIM</b>To evaluate the effects of 60 Hz extremely low frequency (ELF) elelctromagnetic field (EMF) exposure on germ cell apoptosis in the testis of mice.</p><p><b>METHODS</b>Adult male BALB/c mice (7 weeks of age) were exposed to a 60 Hz EMF of 0.1 mT or 0.5 mT for 24 h/day. A sham-exposed group served as the control. After 8 weeks of exposure, the mice were sacrificed. Germ cell apoptosis in the testis was assessed by histopathological examination, the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) and flow cytometric examination of isolated spermatogenic cells stained with 7 aminoactinomycin D (7-AAD).</p><p><b>RESULTS</b>EMF exposure did not significantly affect the body and testis weights, but significantly increased the incidence of germ cell death. The distinguishing morphological feature of EMF exposure was a decrement in the number of well organized seminiferous tubules. Quantitative analysis of TUNEL-positive germ cells showed a significantly higher apoptotic rate in the 0.5 mT exposed mice than that in the sham controls (P<0.05), while the difference between the two exposed groups was insignificant. The TUNEL-positive cells were mainly spermatogonia. In flow cytometry analysis, the percentage of live cells [forward scatter count (FSC)(high)7-AAD(-)] was lower in the exposed groups than that in the controls (Figure 5A), but the decrease in viability was not statistically significant.</p><p><b>CONCLUSION</b>Continuous exposure to ELF EMF may induce testicular germ cell apoptosis in mice.</p>


Subject(s)
Animals , Male , Mice , Apoptosis , Body Weight , Electromagnetic Fields , Flow Cytometry , In Situ Nick-End Labeling , Mice, Inbred BALB C , Organ Size , Spermatozoa , Cell Biology , Testis , Cell Biology
11.
Journal of Bacteriology and Virology ; : 177-181, 2003.
Article in Korean | WPRIM | ID: wpr-40000

ABSTRACT

Leptospirosis has been one of important epidemic diseases in Korea since 1984. Wild rodents, mostly Apodemus agrarius, served the important source of infection especially in harvest season in rural areas of Korea. Prevalence of Leptospira spp. infection in field rodents were investigated by detecting leptospiral DNA from rodent kidney. Among 108 rodents collected from various areas in 1998, leptospiral DNA were detected from 7 rodents (6.48%). Among 104 rodents, Apodemus agrarius, captured from Yeonchon and Paju area in 2001 and 2002, leptospiral DNA were detected from 6 rodents (5.76%). No leptospiral DNA was detected from 23 rodonts (Apodemus peninsulae 16, Apodemus agrarius 2 and Eothenomys regulus 5) captured in Odae mountain area in 1998.


Subject(s)
Animals , DNA , Kidney , Korea , Leptospira , Leptospirosis , Murinae , Polymerase Chain Reaction , Prevalence , Rodentia , Seasons
12.
Journal of Bacteriology and Virology ; : 247-254, 2002.
Article in Korean | WPRIM | ID: wpr-44293

ABSTRACT

Murine typhus is an acute febrile illness caused by Rickettsia typhi. It is one of the four major acute febrile illnesses in Korea during autumn. To study a species-specific antigen of R. typhi, two clinical isolates (87-91 and 87-100) and two reference strains (VR-144 and VR-738) were analyzed by mouse antisera and monoclonal antibodies (MAbs). On SDS- polyacrylamide gel electrophoresis (PAGE), R. typhi showed major antigen bands of 135, 80, 75, 64, 47, 22, and 19 kDa and these bands differed with those of other species. On Western blot analysis, the MAbs reacting only with R. typhi could only detect 135 kDa protein. The 135 kDa protein appeared to be the species-specific antigen. Other MAbs showing cross-reactivity with R. prowazekii reacted with 135 kDa protein in fresh culture supernatant of R. typhi infected host cell. However, the cross-reacting antibody did also react with smaller protein bands, most of which seem to be degradation products of the 135 kDa protein since they increase in old protein stocks purified from R. typhi harvested from infected host cell. These suggest that 135 kDa protein is unstable and the R. typhi specific epitopes are located at the regions of 135 kDa protein that are removed when the protein is degraded. The 135 kDa protein or its specific and stable recombinant protein would serve an important target for the development of vaccine and specific diagnostic antigen.


Subject(s)
Animals , Mice , Antibodies, Monoclonal , Blotting, Western , Electrophoresis, Polyacrylamide Gel , Epitopes , Immune Sera , Korea , Rickettsia typhi , Rickettsia , Typhus, Endemic Flea-Borne
13.
Journal of Korean Orthopaedic Research Society ; : 18-26, 1999.
Article in Korean | WPRIM | ID: wpr-40634

ABSTRACT

We investigated the feasibility of repairing cartilage defect with cultured mesenchymal stem cell from bone marrow. Mesenchyaml stem cells were aspirated from bone marrow of mature rabbits, and cultured in monolayer. Osteochondral defect was artificially made on the patellar groove of distal femur of the same rabbit, and fascia overlying quadriceps was harvested and sutured around the defect, then cultured mesenchymal cells were injected to the defects. Sixteen mature New Zealand white rabbits were used as experimental group and 13 rabbits were used as control. The rabbits were killed at after 14 weeks. Healing of defect was investigated histologically with H-E and toluidine blue staining, biochemically with immunohistochemical staining for Type II collagen, and also at molecular level through RT-PCR for mRNA of type II collagen. Semiquantitative histological scores were significantly higher in experimental group than thoese in control(P<0.05). Immunohistochemical staining showed high signal thor type II collagen on newly formed cartilage in the defect. RT-PCR detected mRNA for type II collagen from mature chondrocytes. Our data suggest that repair of cartilage defect can be enhanced with transplantation of cultured mesenchymal stem cell.


Subject(s)
Rabbits , Bone Marrow , Cartilage , Chondrocytes , Collagen Type II , Fascia , Femur , Mesenchymal Stem Cells , RNA, Messenger , Stem Cells , Tolonium Chloride
14.
Journal of the Korean Society for Microbiology ; : 409-421, 1999.
Article in Korean | WPRIM | ID: wpr-154331

ABSTRACT

Serovars of 22 leptospiral field isolates from rats trapped in Korea were identified by cross-agglutinin absorption test (CAAT). Genomic characteristics of 7 selected isolates and 6 antigenically closely related reference serovars of lai, yeonchon, birkini, gem, mwogolo, and canicola were differentiated by arbitrarily primed PCR (AP-PCR) and southern blot hybridization using 16S rRNA gene probe from Borrelia burgdorferi. Among the 22 isolates, 21 strains were identified as serovar lai by CAAT, while the serological reactivity of NR13 did not accord with that of serovar lai. Results of AP-PCR using primers RSP, KF and PB-1 were in general agreement with those obtained by serological identification, and all 7 isolates including NR13 showed the same profile with serovar lai or yeonchon. In the southern blot hybridization with 16S rRNA gene probe, the isolates were divided into two ribotype groups when HindIII and BamHI digests were employed: isolates NR4, NR13, and serovar lai showed the same profile, and isolates JR34, JR57, KR48, JR77, and JR82 were classified as the another ribotype group. Isolate NR13 and serovar yeonchon, which were isolated in Korea and showed serological differences with serovar lai, were indistinguishable from serovar lai in this DNA study using AP-PCR and ribotyping. These results demonstrate that Korean leptospiral isolates were closely related in DNA level, and ribotyping would be useful for subgrouping of field isolates.


Subject(s)
Animals , Rats , Absorption , Blotting, Southern , Borrelia burgdorferi , DNA , Genes, rRNA , Korea , Leptospira , Polymerase Chain Reaction , Ribotyping
15.
Journal of the Korean Society for Microbiology ; : 591-594, 1999.
Article in Korean | WPRIM | ID: wpr-220176

ABSTRACT

Leptospirosis has been known as endemic disease in Korea since 1984. Wild rodent, mostly Apodemus agrarius, has been known as an important source of leptospiral infection especially in rainy circumstances in harvest reason of rural area. The infection rates of Leptospira interrogans in field rodents, Apodemus agrarius, was investigated by culture and PCR detection of leptospiral DNA, and compared with previous data. Furthermore, the serogroup and serovar were investigated. Two hundred twenty two Apodemus agrarius were captured during October to December 1996. Spirohaetes were isolated from 22 (9.9%) and leptospiral DNA was detected in an additional six rodents (12.6%). Subsequent cross-agglutinin absorption test, monoclonal antibody reactivity classified 21 cultures among 22 isolates as Leptospira interrogans serogroup Icterohemorrhagiae serovar lai. The above data did not differ from previous survey in 1984 to 1987. There was no significant change of Leptospira interrogans infection in field rodents in Korea.


Subject(s)
Animals , Absorption , DNA , Endemic Diseases , Korea , Leptospira interrogans , Leptospira , Leptospirosis , Murinae , Polymerase Chain Reaction , Prevalence , Rodentia
16.
Korean Journal of Nuclear Medicine ; : 250-258, 1998.
Article in Korean | WPRIM | ID: wpr-66419

ABSTRACT

PURPOSE: ATP (adenosine triphosphate) is a potent coronary vasodilator with a rapid onset of action and a very short half-life. Myocardial perfusion scintigraphy with intravenous ATP has not yet been sufficiently proven in the diagnosis, follow-up, and risk stratification of coronary artery disease. The purpose of this study was to evaluate the safety, feasibility and diagnostic accuracy of pharmacologic stress thallium-201 myocardial SPECT using an intravenous ATP infusion in patients with suspected coronary artery disease. MATERIALS AND METHODS: Thalliurn-201 myocardial SPECT in 319 patients with suspected coronary artery disease were performed after the infusion of ATP (0.08 mg/kg/min for 6 rnin). The adverse effects were carefully monitored. Coronary angiography was also performed within 3 weeks. RESULTS: Although 76.5% of the patients had sorne adverse effects, they were transient, mild, and well tolerated. In all patients, the ATP infusion protocol was completed and only 2 patients required aminophylline. The adverse effects were dyspnea in 63%, headache in 31%, flushing in 21%, chest pain in 14% and abdominal discomfort in 5% of the patients. The sensitivity and specificity were 80% and 90% respectively. CONCLUSION: Thallium-201 myocardial SPECT after 6 min-infusion of ATP at a rate of 0.08 mg/kg/min is safe and has a diagnostic value in detecting coronary artery disease.


Subject(s)
Humans , Adenosine Triphosphate , Aminophylline , Chest Pain , Coronary Angiography , Coronary Artery Disease , Coronary Vessels , Diagnosis , Dyspnea , Flushing , Follow-Up Studies , Half-Life , Headache , Infusions, Intravenous , Perfusion Imaging , Sensitivity and Specificity , Tomography, Emission-Computed, Single-Photon
17.
Journal of the Korean Society of Virology ; : 53-62, 1998.
Article in English | WPRIM | ID: wpr-17126

ABSTRACT

Human papillomavirus (HPV) 16, E7 proteins derived from the prototype (Bac73) and natural variant (Bac101) E7 open reading frame were produced in Sf9 insect cells. The variant E7 gene occurred naturally by substitution mutation at the position of 88 nucleotide, resulting serine instead of asparagine. Using E7 specific monoclonal antibody (VD6), both E7 proteins were identified in recombinant baculovirus infected SF9 cells. Radiolabelling and immunoprecipitation analysis revealed that both E7 proteins were phosphoproteins. Immunostaining result showed that E7 proteins were mainly localized in the cytoplasm. Nuclear form of E7 proteins was also detected after a sequential fractionation procedure for removing chromatin structure. Considering that the VD6 recognition site in E7 protein is located within 10 amino acid at the N-terminus, this region appears to be blocked by the nuclear component. Western blot analysis revealed that nuclear form was more abundant than cytoplasmic E7 proteins. Time course immunostaining showed that the primary location of E7 protein was the nucleus and exported to the cytoplasm as proteins were accumulated. These events occurred similarly in both Bac73 and Bac101 infected Sf9 cells, suggesting that these two proteins may have similar biological functions.


Subject(s)
Humans , Asparagine , Baculoviridae , Blotting, Western , Chromatin , Cytoplasm , Immunoprecipitation , Insecta , Open Reading Frames , Phosphoproteins , Serine , Sf9 Cells
18.
Korean Journal of Otolaryngology - Head and Neck Surgery ; : 1267-1273, 1997.
Article in Korean | WPRIM | ID: wpr-645609

ABSTRACT

BACKGROUND: Heat shock proteins(HSPs) are released from the cells by temperature elevation or other stresses, including cytokines, hypoxia, inflammation or reactive oxygen species. These proteins likely play a role in cellular repair and survival mechanisms. OBJECTIVES: In order to elucidate the relationship between the HSP 70 and viral induced maxillary sinusitis. MATERIALS AND METHODS: The degree of HSP expression was evaluated in New Zealand white rabbits with maxillary sinusitis induced by inoculation of Influenza A virus(KOREA /1/ 80/H3N2) into the mucosa. The animals were serially sacrificed 1, 5, 7, 14, 21, 28th day after inoculation. The localization of the induced form of HSP 70 in the normal & infected maxillary mucosa were studied with immunohistochemical staining. RESULTS: The normal sinus mucosa did not show any staining for the HSP 70. As contrasted with the normal group, the mucosa of the first day after viral inoculation showed very light staining in the epithelial layer. The degree of immunoreactive staining was gradually increased up to the seventh day. Epithelial layer of the mucosa, cilia and submucosa were heavily stained at seventh day after inoculation, and then the degree was reduced at 14th day. The staining of the mucosa was completely disappeared at 28th day after viral inoculation. CONCLUSION: These results suggest that the HSP 70 was produced from acute stage of the infected mucosa by the Influenza A virus. Presumably, it is closely related to the inflammatory reaction in the mucosa of the maxillary sinus.


Subject(s)
Animals , Rabbits , Hypoxia , Cilia , Cytokines , Heat-Shock Proteins , Hot Temperature , HSP70 Heat-Shock Proteins , Inflammation , Influenza A virus , Influenza, Human , Maxillary Sinus , Maxillary Sinusitis , Mucous Membrane , Reactive Oxygen Species , Shock
19.
Journal of the Korean Society for Microbiology ; : 335-342, 1997.
Article in Korean | WPRIM | ID: wpr-100824

ABSTRACT

The gene encoding E7 oncoprotein of human papillomavirus type 16 was cloned and expressed in Escherichia coli, and the monoclonal antibodies (Mabs) against this expressed protein were generated. For the efficient immunization, two kind of recombinant E7 protein in fusion form were produced. One was maltose binding protein (MBP) fusion type (MBP-E7) and the other was T7 phage gene 10 product fusioa type (gene 10-E7). Immunization with these two fusion protein to mice, finally two Mabs (VD6 and IB10) were obtained. VD6 and IB10 showed reactivities with E7 protein in CaSki cell but not in HeLa by Western blot analysis. In addition, the Mab, VD6, reacted with COS-7 cell transfected with E7 gene majorly in cytoplasm by immunofluorescence test. Also VD6 could detect E7 protein in cytoplasm and nucleus of CaSki ceU by immunogold electron microscopy. Based on these results, the Mab VD6 was could be used for various E7 detection system such as Western blot analysis and immunohistochemical methods.


Subject(s)
Animals , Humans , Mice , Antibodies, Monoclonal , Bacteriophage T7 , Blotting, Western , Clone Cells , COS Cells , Cytoplasm , Escherichia coli , Fluorescent Antibody Technique , Immunization , Maltose-Binding Proteins , Microscopy, Electron
20.
Journal of the Korean Radiological Society ; : 927-931, 1995.
Article in Korean | WPRIM | ID: wpr-41190

ABSTRACT

PURPOSE: To evaluate the chest radiographic findings of rickettsial disease including murine typhus and tsutsugamushi disease in Chunchon. MATERIALS & METHODS: Chest radiographic films of 81 cases diagnosed as rickettsial disease(55 cases of tsutsugamushi disease, 26 cases of murine typhus) by immunofluorescence test were retrospectively analyzed. RESULTS: Main serotypes of Rickettsia tsutsugamushi were Gilllain and Karp. Incidence rate of tsutsugamushi disease was 2.1 times greater than that of murine typhus. Chest radiographs were abnormal in 63.6% of tsutsugamushi disease, and in 30.8% of murine typhus. Radiographic findings were Kerly's B line, reticu-Ionodular densities, hilar enlargement, pleural effusion, and splenomegaly in both entities, but pulmonary consolidation was only found in tsutsugamushi disease. The patients with the abnormal radiographic findings were statistically well correlated with cardiomegaly(p<0.01) and azygos engorgement(p<0.05), as compared to the patients with normal radiographic findings. CONCLUSION: Radiographic findings of both murine typhus and tsutsugamushi disease were interstitial pattern. But the chest radiographs in patients with tsutsugamushi disease showed more severe pattern with higher rate of abnormality.


Subject(s)
Humans , Fluorescent Antibody Technique , Incidence , Orientia tsutsugamushi , Pleural Effusion , Radiography, Thoracic , Retrospective Studies , Scrub Typhus , Splenomegaly , Thorax , Typhus, Endemic Flea-Borne
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