ABSTRACT
BACKGROUND AND OBJECTIVES: A heparin-binding polypeptide called midkine is a family of secreted growth/differentiation cytokines and has a role in tumor growth by enhancing endothelial proliferation, vascular density and angiogenesis. In this respect, midkine may be involved in the pathogenesis of nasal polyposis. The aim of the present study is to evaluate the expression of midkine mRNA in the human nasal mucosa and polyps. MATERIALS AND METHOD: The total RNA was isolated from freshly disected inferior turbinate of patients who underwent rhinoplasty and from nasal polyps of chronic rhinosinusitis patients. The expression and distribution of midkine mRNA was investigated by reverse transcriptse- polymerase chain reaction (RT-PCR) and in situ hybridization. The midkine mRNA expression in nasal mucosa and polyps were semi-quantitatively evaluated by Southern blot hybridization. RESULTS: The expression of midkine mRNA was identified in both normal inferior turbinate and nasal polyp. Histochemistry of in situ hybridization revealed that midkine mRNA in normal inferior turbinate was intensely expressed in the surface epithelium, submucosal glands, vascular endothelium, and inflammatory cells scattered in submucosal tissues. Midkine mRNA was expressed in the nasal polyps, many inflammatory cells and newly formed vascular endothelium, but not in the newly formed glandular epithelium. In semi-quantitative southern blot hybridization, midkine mRNAs did not have different expression levels between inferior turbinate and nasal polyps. CONCLUSION: These results indicate that midkine mRNA is innately expressed in human nasal mucosa, playing a role in nasal physiology. Also, the results show that midkine may be involved in the pathogenesis of nasal polyps via angiogenesis, tissue growth, and inflammatory process.
Subject(s)
Humans , Blotting, Southern , Cytokines , Endothelium, Vascular , Epithelium , In Situ Hybridization , Nasal Mucosa , Nasal Polyps , Physiology , Polymerase Chain Reaction , Polyps , Rhinoplasty , RNA , RNA, Messenger , TurbinatesABSTRACT
BACKGROUND AND OBJECTIVES: To analyze results of treatment and prognostic factors in patients with T3 supraglottic carcinoma and to compare results of treatment in patients with T3 transglottic carcinoma with T3 pure supraglottic carcinoma. MATERIALS AND METHOD: A retrospective study was done on thirty-two patients who underwent a surgery or surgery with postoperative radiation therapy from 1990 to 2000. Neck dissection was performed in 27 patients and 24 patients received postoperative radiation therapy. RESULTS: The 3-year overall survival rate was 81.6%. The 3-year overall survival rate of T3 pure supraglottic carcinoma and T3 transglottic carcinoma were 91.7% and 73.2%, respectively (p<0.05). The univariate analysis revealed a prognostic significance for vocal cord fixation and statistical trend to age, dyspnea, clinical and pathological metastasis of cervical lymph node and postoperative radiation therapy (p<0.2). T3 transglottic carcinoma was significantly correlated with vocal cord fixation. CONCLUSION: Surgery or surgery with postoperative radiation therapy provides acceptable rates of cancer control and survival rate for patients with T3 supraglottic carcinoma. Transglottic involvement and vocal cord fixation shown by the fiberoptic laryngoscopy were significant prognostic factors. T3 transglottic cancer needs more aggressive management.
Subject(s)
Humans , Dyspnea , Laryngoscopy , Lymph Nodes , Neck Dissection , Neoplasm Metastasis , Prognosis , Retrospective Studies , Survival Rate , Vocal CordsABSTRACT
BACKGROUND: Epstein-Barr virus (EBV) is causative agent of infectious mononucleosis and nasopharyngeal carcinoma and associated with Burkitt lymphoma and other tumors. The recombinant protein is needed for the rapid and sensitive serodiagnosis of EBV infection. METHODS: EBV gene encoding the protein reactive with the sera of EBV-infected patient was cloned and characterized with lambda gt11 expression library of cDNA of EBV B95-8 strain. RESULTS: The recombinant proteins from clone 12, 15 and 21 were expressed as 120, 118, 160 kDa-usion protein with beta-galactosidase, respectively, which were reactive with IgG anti-EBV antibody-positive sera, but not with anti-EBV antibody-negative sera. A homology search of the DNA sequence of the cloned gene with EBV B95-8 sequences revealed that those were located at 61716~62087, 61898~62085, and 102128~103158, respectively. These positions correspond to BFRF3, BFRF3, and BZLF1, respectively, which were reported as immunogenic protein which evoked strong and longstanding antibody response and had no sequence homology with the proteins of other herpesviruses. All the patients' sera were reactive with clone 12 protein, but only 5 out of 9 patients' sera were reactive with clone 21 protein. CONCLUSION: Clone 21 protein expressing BFRF3 fragment was immunoreactive in patient sera from natural EBV infection and was regarded as useful candidate for the serodiagnosis of EBV infection.
Subject(s)
Humans , Antibody Formation , Base Sequence , beta-Galactosidase , Burkitt Lymphoma , Clone Cells , Cloning, Organism , DNA, Complementary , Epstein-Barr Virus Infections , Herpesviridae , Herpesvirus 4, Human , Immunoglobulin G , Infectious Mononucleosis , Recombinant Proteins , Sequence Homology , Serologic TestsABSTRACT
Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CDl4 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with 10 microM, 50 microM or 100 microM of TPA. The cell morphology change was observed and the expression of the CD11b and CDl4 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 h and 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.
Subject(s)
Humans , Binding Sites , Cytomegalovirus , Fireflies , Luciferases , Macrophages , Permissiveness , Transfection , U937 CellsABSTRACT
Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CDl4 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with 10 microM, 50 microM or 100 microM of TPA. The cell morphology change was observed and the expression of the CD11b and CDl4 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 h and 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.
Subject(s)
Humans , Binding Sites , Cytomegalovirus , Fireflies , Luciferases , Macrophages , Permissiveness , Transfection , U937 CellsABSTRACT
Human cytomegalovirus (HCMV) glycoprotein H (gH) is one of target molecules in the human immune response to HCMV infection. This study was performed to rneasure the immune responses to HCMV gH in human sera by the expression of HCMV gH in eukaryotic cells. Amplified DNA from the gene encoding gH of HCMV by polymerase chain reaction was cloned into pcDNA3 to construct eukaryotic expression vector. Immunofluorescent staining revealed that the expressed gH in mammalian cells was reactive with the specific monoclonal antibody. Antibody titer in patient's sera with HCMV infection was measured with HCMV-infected fibroblasts and HCMV gH expressed in mammalian cells. Anti-HCMV gH antibody titer was higher in patient group than in healthy control group. There was no correlation between the antibody titer to the whole HCMV and neutralizing antibody titer, and between the antibody titer to whole HCMV and whole gH. Conclusively it is highly recommendable to use the defined antigen such as HCMV gH for the detection of antibody in HCMV-infected persons in the aspect of immunological properties.
Subject(s)
Humans , Antibodies, Neutralizing , Clone Cells , Cytomegalovirus , DNA , Eukaryotic Cells , Fibroblasts , Glycoproteins , Polymerase Chain ReactionABSTRACT
Human cytomegalovirus (HCMV) has the ability to activate the espremission of many viral and cellular genes. Among various viral proteins, the immediate early proteins (IE1-72kDa, IE2-86kDa) have been known to be potent transactivators. The product of c-jun photo-oncogene is important in cell activation and differentiation. Here, we tried to find out if the IE could activate the c-jun promoter and also tried to identify the responsible sequence elements in the c-jun activation by IE1-72kDa. We found HCMV IE expression transactivated the c-jun promoter in human embryonal lung fibroblasts (HEL). The activation fold by IE1-72kDa, IE2-86kDa and IE2-55kBa was 23, 35, and 5, respectively. When the expression of each IE was combined, it showed synergism. Expression of (IE1-72kDa + IE2-86kBa) and (IE1-72kDa + IE2-86kDa + IE2-55kDa) resulted in 131 and 162 fold increase, respectively. The c-jun promoter region between -117 and -59 contains binding sites for the transcription factors Spl, CAAT, AP-1 like (ATF/CREB), and MEF2. Transient expression assays were performed using various reporter plasmids containing the c-jun promoter-regulatory region linked to the luciferase gene and a plasmic expressing HCMV IE1 gene. Deletional and point mutational analysis showed that the sequence between -225 to -160 and the CTF binding site were involved in the up-regulation of c-jun promoter.
Subject(s)
Humans , Binding Sites , Cytomegalovirus , Fibroblasts , Immediate-Early Proteins , Luciferases , Lung , Plasmids , Promoter Regions, Genetic , Trans-Activators , Transcription Factor AP-1 , Transcription Factors , Up-Regulation , Viral ProteinsABSTRACT
PURPOSE: Recently, it is suggested that the inhibitian of apoptosis is associated with tumorigenesis of colon. Bcl-2 gene is an important inhibitory regulator of apoptosis, and bcl-2 acts antagonistly with the wild type p53 gene, one of the tumor suppressor genes, in apoptosis. To detnmine the role of bcl-2 and p53 gene in colonic tumorigenesis, we performed the study. MATERIALS AND METHODS: We tested the tissue obtained by polypectomy and surgical resection by immunohistochemical staining for Bcl-2 and p53. RESULTS: We found that in normal colonic tissue, the Bcl-2 was sparcely expressed, and the p53 was expressed sporadically. The rate of positivity of staining was below 5%. However, in colonic adenoma and colon cancer tissue, Bcl-2 and p53 were expressed more than in nonnal colonic tissue(p<0.05). (Scoring in Colonic adenoma: Bcl-2 6.2+/-1.1, p53 5.7+/-1.0; Scoring in Colonic carcinoma: Bcl-2 4.7+/-1.0, p53 8.3+/-0.9) CONCLUSION: Our results suggested that the bcl-2 and p53 play an important role in colonic tumorigenesis.
Subject(s)
Adenoma , Apoptosis , Carcinogenesis , Colon , Colonic Neoplasms , Genes, bcl-2 , Genes, p53 , Genes, Tumor SuppressorABSTRACT
Adenoviruses are icosahedral virions containing double-stranded linen DNA. They are 70 nm to 90 nm in diameter and capsid is composed of 252 capsomeres. Several members of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be oncogenic in young rat. Previous report involving effect of caffein on transformation induced by Adenovirus type 12 [9] has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the of(tract of MNNG (N-methyl-N'-nitro-N-nitroguanidine) on transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigate the effect of MNNG on the adenovirus type 12-induced transformation in L cell. Results were as following 1. Adeno virus type 12 induced transformation was enhanced in the presence of MNNG. 2. Yields of adeno type 12 virus in L cell were slightly inhibited by treatment of MNNG.
Subject(s)
Animals , Cricetinae , Rats , Adenoviridae , Bedding and Linens , Capsid , DNA , Methylnitronitrosoguanidine , VirionABSTRACT
The human cytomegalovirus(HCMV) gene encoding the protein reactive with the sera of HCMV-infected patient was cloned and characterized. A reactive phage clone was screened from a lambda gt11 expression library of cDNA of HCMV AD169 strain using HCMV-infected patient sera. The recombinant protein was expressed as 138 kDa-fusion protein with beta-galactosidase, which was reactive with IgM or IgG HCMV antibody-positive sera, but not with anti-HCMV antibody-negative sera. A homology search of the DNA sequence of the cloned gene with HCMV AD169 sequences revealed that it was composed of 709 base pairs spanning between 0.174 and 0.177 map units of the UL32 region of the HCMV AD169 strain genome. This position corresponded to a part of the gene encoding 150 kDa phosphoprotein-(pp150), a major tegument protein, which was reported as an immunogenic protein which evoked strong and longstanding antibody response and had no sequence homology with the proteins of other herpesviruses. These results suggested that pp150 was an immunogenic protein in natural HCMV infection and therefore this clone was regarded as a useful candidate for developing an antigen for the serodiagnosis of HCMV.
Subject(s)
Humans , Antibodies, Viral/blood , Antigens, Viral/genetics , Cloning, Molecular , Cytomegalovirus/genetics , Cytomegalovirus Infections/blood , DNA, Complementary/genetics , DNA, Viral/genetics , Gene Library , Genes, Viral , Recombinant Fusion Proteins/biosynthesis , Sequence Homology, Nucleic Acid , Viral Matrix Proteins/geneticsABSTRACT
Twenty-four monoclonal antibodies were produced by immunizing BALB/c mice with Rickettsia tsutsugamushi Boryong strain and used for the analysis of antigenic characteristics of R.tsutsugamushi Boryong strain and antigenic heterogeneity of R.tsutsugamushi by indirect immunofluorescent(IF) test. R. tsutsugamushi Kato, Karp, Gilliam, TA686, TA716, TA763, TC586, TH1817, and Boryong were used for the analysis of antigenic heterogeneity of R.tsutsugamushi. Five monoclonal antibodies were reactive with 27-kDa protein, four monoclonal antibodies were reactive with 47-kDa protein, and eight monoclonal antibodies were reactive with 56-kDa protein of R.tsutsugamushi Boryong strain. The reactive protein of seven monoclonal antibodies could not be identified by immunoblotting method. All monoclonal antibodies to 27-kDa protein and three monoclonal antibodies to 47-kDa protein, and five monoclonal antibodies to 56-kDa protein were reactive with three to eight strains among nine strains of R. tsutsugamushi tested. One monoclonal antibody reactive to 47-kDa protein(KI18) and two monoclonal antibodies reactive to 56-kDa protein(KI36, and KI37) reacted with all the strains of R. tsutsugamushi tested. Strain-specific monoclonal antibody(KI58) could be found among antibodies which were reactive with 56-kDa protein. There was no strain which showed same reactivity pattern to these 24 monoclonal antibodies among nine strains. From this results, it could be concluded that Boryong strain is antigenically different from other strains of R.tsutsugamushi and antigenic heterogeneity of R.tsutsugamushi is due to the antigenic diversity of several proteins of R. tsutsugamushi including 56-kDa protein.
Subject(s)
Animals , Mice , Antibodies, Monoclonal/immunology , Antigens, Bacterial/analysis , Bacterial Proteins/analysis , Mice, Inbred BALB C , Orientia tsutsugamushi/immunology , Species SpecificityABSTRACT
No abstract available.