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1.
Auris Nasus Larynx ; 37(2): 137-44, 2010 Apr.
Article in English | MEDLINE | ID: mdl-19505782

ABSTRACT

OBJECTIVES: The aim of this study is to examine the internalization of nontypeable Haemophilus influenzae (NTHi) into human epithelial cells. METHODS: Bactericidal assay was applied to examine the effects of antibiotics against cell-adherent NTHi using HEp-2 cells. A trans-well chamber assay was applied to examine the internalization and penetration of NTHi using Detroit562 cells. RESULTS: The adherence of NTHi to HEp-2 cells was noted after 2h of incubation. Azithromycin had a strong bactericidal effect against both cell-associated and non-adherent NTHi, while ceftriaxone did not show bactericidal effects on NTHi adhered to the HEp-2 cells. Three (60.0%) out of five NTHi isolates from the nasopharynx of children with intractable acute otitis media (AOM) internalized into and subsequently penetrated through the epithelial cells at various degrees. Azithromycin had a strong bactericidal effect against the cell-internalized NTHi, while ceftriaxone was bactericidal only against extracellular NTHi. CONCLUSION: The potential of NTHi as the intracellular pathogen may contribute to the persistent existence of this pathogen that result in the prolonged and intractable clinical course of AOM. Azithromycin may be a therapeutically significant antibiotic for patients with prolonged respiratory tract infections due to NTHi.


Subject(s)
Anti-Bacterial Agents/pharmacology , Azithromycin/pharmacology , Bacterial Adhesion/drug effects , Bacterial Physiological Phenomena/drug effects , Ceftriaxone/pharmacology , Epithelial Cells/microbiology , Gentamicins/pharmacology , Haemophilus Infections/microbiology , Haemophilus influenzae/classification , Haemophilus influenzae/isolation & purification , Otitis Media with Effusion/microbiology , Acute Disease , Cell Line , Child , Dose-Response Relationship, Drug , Epithelial Cells/drug effects , Haemophilus Infections/drug therapy , Haemophilus influenzae/drug effects , Humans , In Vitro Techniques , Microbial Sensitivity Tests , Microbial Viability/drug effects , Otitis Media with Effusion/drug therapy
2.
Auris Nasus Larynx ; 36(5): 578-85, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19268507

ABSTRACT

OBJECTIVE: We aimed to define role of tonsillar lymphocytes (TL) and immune cross-reactivity between bacterial-HSP65 and human-HSP60 in Pustulosis palmaris et plantaris (PPP), an intractable chronic disease characterized with pustules and cornification of palms and soles. METHODS: Two sets of crossover trials were designed by employing SCID mice model. In the first trial, mice were transplanted with tonsillar lymphocytes and skin-grafts from PPP patients (TL group). In the second trial, mice were transplanted with tonsillar lymphocytes from PPP patients and injected with recombinant human HSP60. Control groups were designed for each step. Comparisons were performed for immunologic analyses including infiltration of CD4+ lymphocytes in skin-grafts by immunostaining, and levels of anti-HSP65-IgG and cytokines in mice sera by enzyme-linked immunosorbent assay (ELISA). RESULTS: In TL group, infiltration of CD4+ lymphocytes in skin-grafts were significantly higher than mice transplanted with blood lymphocytes (p<0.05), while anti-HSP65-IgG levels in sera showed non-significant tendency to increase in the TL group. CD4+ cells and anti-HSP65-IgG levels were also well-correlated with each other in TL group (p<0.01). Besides, anti-HSP65-IgG levels were significantly correlated with cytokine levels (IL-6, IFN-gamma) in mice sera (p<0.01). We found strong expression of HSP60 in PPP lesions. Finally, HSP60-stimulation in mice transplanted with TL from PPP patients induced significantly higher anti-HSP65-IgG levels in serum compared to control groups including mice without HSP60-stimulation or peripheral blood lymphocytes-transplanted mice or transplanted with TL from control patients (p<0.05). CONCLUSION: Our results indicate the pathogenic role of TL and immune cross-reaction between human-HSP60 and bacterial-HSP65 in PPP.


Subject(s)
Bacterial Proteins/immunology , Chaperonin 60/immunology , Cross Reactions , Heat-Shock Proteins/immunology , Lymphocytes/immunology , Palatine Tonsil/immunology , Psoriasis/immunology , Adult , Animals , Antibodies/blood , Biopsy , CD4 Antigens/metabolism , CD4 Lymphocyte Count , Chaperonin 60/metabolism , Chimera , Cross-Over Studies , Cytokines/blood , Epithelium/metabolism , Epitopes , Female , Humans , Immunoglobulin G/blood , Male , Mice , Mice, SCID/blood , Middle Aged , Palatine Tonsil/pathology , Psoriasis/metabolism , Psoriasis/pathology , Skin/metabolism , Skin/pathology , Skin Transplantation , Tonsillectomy , Transplantation, Heterologous
3.
Otolaryngol Head Neck Surg ; 137(1): 119-25, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17599578

ABSTRACT

OBJECTIVES: We sought to assess loss of heterozygosity (LOH) profiles of 3p, 6q, 8q, 10q, 12q, 13q, and 17p and to identify the tumor suppressor genes involved in salivary gland neoplasms. STUDY DESIGN: LOH analysis was performed using 26 microsatellite markers by polymerase chain reaction-polyacrylamide gel electrophoresis method in 20 benign and 6 malignant salivary gland tumors. RESULTS: Overall, LOH was detected in at least one informative locus in 18 of 20 (90%) of benign tumors and in all of 6 cases of malignant tumors. High LOH frequencies were revealed at the loci D3S1307 (22%, 3p26), D3S966 (41%, 3p21), D6S255 (27%, 6q25), D8S166 (25%, 8q12), D8S199 (21%, 8q24), and D10S1765 (28%, 10q23) in benign tumors, defining the hotspot regions for putative tumor suppressor genes. CONCLUSIONS AND SIGNIFICANCE: The hotspot regions defined by the present study suggest that new tumor suppressor genes related to the development of salivary gland tumors may reside at several chromosomal loci, including loci at 3p, 6q, 8q and 10q.


Subject(s)
Chromosomes, Human, Pair 3/genetics , Loss of Heterozygosity/genetics , Parotid Neoplasms/genetics , Submandibular Gland Neoplasms/genetics , Adenolymphoma/genetics , Adenoma, Pleomorphic/genetics , Adult , Aged , Carcinoma/genetics , Carcinoma, Squamous Cell/genetics , Chromosome Mapping , Chromosomes, Human, Pair 10/genetics , Chromosomes, Human, Pair 12/genetics , Chromosomes, Human, Pair 13/genetics , Chromosomes, Human, Pair 17/genetics , Chromosomes, Human, Pair 6/genetics , Chromosomes, Human, Pair 8/genetics , Female , Genes, Tumor Suppressor , Humans , Male , Microsatellite Repeats/genetics , Middle Aged
4.
J Cancer Res Clin Oncol ; 132(1): 19-27, 2006 Jan.
Article in English | MEDLINE | ID: mdl-16170569

ABSTRACT

PURPOSE: Aims of the study are to narrow-down the hotspot region on 10q21 defined by previous genome-wide loss of heterozygosity (LOH) analysis in head and neck squamous cell carcinomas (HNSCC) and to define candidate tumor suppressor genes (TSG) concerned with 10q21. MATERIALS AND METHODS: LOH analysis was carried out with ten polymorphic microsatellite markers. Expression analysis was performed by semi-quantitative RT-PCR, and mutation analysis by PCR and direct sequencing. RESULTS: LOH analysis on 10q21 in 52 HNSCC indicated distinctive and frequent allelic loss at D10S589 (42%). Among flanking genes, we found the RHOBTB1 gene as a candidate TSG, since an intragenic marker demonstrated the highest LOH (44%). Expression analysis revealed down-regulation of RHOBTB1 mRNA in 37% of tumors. Interestingly, all the five tumors that showed decreased expression of RHOBTB1 were accompanied with LOH, supporting the haploinsufficiency and class 2 TSG characteristics of RHOBTB1. No pathogenic mutation of RHOBTB1 was found. Furthermore, another gene within the region, EGR2, was also taken under scope. LOH frequencies around the EGR2 gene were relatively low (23 and 33%). Albeit semi-quantitative expression analysis of EGR2 demonstrated downregulation in 45% of tumor samples, no relation was found between the expression levels and LOH status. CONCLUSION: Frequent allelic loss and decreased expression of RHOBTB1 suggested that this gene has a role in tumorigenesis of a subset of HNSCC.


Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 10 , Early Growth Response Protein 2/genetics , Genes, Tumor Suppressor , Head and Neck Neoplasms/genetics , Loss of Heterozygosity , Base Sequence , DNA Mutational Analysis , Down-Regulation , Exons , Gene Expression Regulation, Neoplastic , Genetic Markers , Humans , Microsatellite Repeats , Molecular Sequence Data , Polymorphism, Genetic , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction
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