Rapid communication: sequence of a quail calpain cDNA.
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Biomedical subjects
Publications and source records attributed to M Ushikai.
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Induction of phase II enzymes such as NADPH:quinone oxidoreductase (QR) can reduce carcinogen-induced mutagenesis and tumor formation. In our search for novel dietary anticarcinogens, fisetin, a flavonol widely distributed in fruits and vegetables was found to induce QR activity in murine hepatoma 1c1c7 cells. The cells were treated with various concentrations of fisetin, and then were assessed for cell growth, QR activity, QR mRNA expression and transcription activation of the QR gene. The results showed that fisetin induced QR activity in time- and dose-dependent manner in the concentration range of 0.1 to 10 microM, and the activity induction was associated with QR mRNA expression as detected by reverse transcription-PCR. Furthermore, transfection studies using a human QR antioxidant/electrophile-response element (ARE/EpRE) reporter construct demonstrated that fisetin activated the ARE/EpRE. These results show that fisetin increases QR activity by transcriptional activation of the ARE/EpRE, suggesting a novel mechanism by which dietary fisetin may be implicated in cancer chemoprevention.
Penicillin-resistant Streptococcus pneumoniae (PRSP) is a frequently detected pathogen of intractable acute otitis media and is associated with prolonged or recurrent infection. The use of antibiotics has made the incidence of secondary acute mastoiditis following acute otitis media relatively rare, but when it does occur, its severe complications may be life-threatening. We report a case of pediatric recurrent acute mastoiditis caused by PRSP in a 6-year-old boy suffering from PRSP acute mastoiditis on 4 occasions, twice undergoing simple mastoidectomy. Although we initially suspected PRSP to be the chief factor in iterative infection, immunological analysis demonstrated significantly decreased IgG and IgA antibodies in serum and the patient was diagnosed as having common variable immunodeficiency (CVID). As the first middle ear infection occurred at the age of 6 and there was no history of upper respiratory tract infection, CVID may be the main pathological factor of recurrent mastoiditis, although infection occurred, only in the ear and did not involve other organs. This suggests that recurrent mastoiditis in the present case involved the coexistence of PRSP and CVID.
Adult-onset type II citrullinemia (CTLN2) is characterized by a liver-specific deficiency of argininosuccinate synthetase (ASS) protein. We have recently identified the gene responsible for CTLN2, viz., SLC25A13, which encodes a calcium-binding mitochondrial carrier protein, designated citrin, and found five mutations of the SLC25A13 gene in CTLN2 patients. In the present study, we have identified two novel mutations, 1800ins1 and R605X, in SLC25A13 mRNA and the SLC25A13 gene. Diagnostic analysis for the seven mutations in 103 CTLN2 patients diagnosed by biochemical and enzymatic studies has revealed that 102 patients had one or two of the seven mutations and 93 patients were homozygotes or compound heterozygotes. These results indicate that CTLN2 is caused by an abnormality in the SLC25A13 gene, and that our criteria for CTLN2 before DNA diagnosis are correct. Five of 22 patients from consanguineous unions have been shown to be compound heterozygotes, suggesting a high frequency of the mutated genes. The frequency of homozygotes is calculated to be more than 1 in 20,000 from carrier detection (6 in 400 individuals tested) in the Japanese population. We have detected no cross-reactive immune materials in the liver of CTLN2 patients with any of the seven mutations by Western blot analysis with anti-human citrin antibody. From these findings, we hypothesize that CTLN2 is caused by a complete deletion of citrin, although the mechanism of ASS deficiency is still unknown.
OBJECTIVE/METHODS: Long-term administration of clarithromycin has been reported to be effective in the treatment of chronic sinusitis. To investigate the mechanism underlying the anti-inflammatory activity of clarithromycin, the authors evaluated the effect of clarithromycin on the gene expression of proinflammatory cytokine and the DNA-binding activity of nuclear factor (NF)-kappa B in cultured human nasal epithelial cells and fibroblasts. Cells were incubated with endotoxin purified from nontypable Haemophilus influenzae or interleukin (IL)-1 beta in the presence of clarithromycin. RESULTS: Northern blot analysis revealed that clarithromycin suppressed IL-1 beta gene expression in human nasal epithelial cells stimulated by H. influenzae endotoxin (HIE). Intercellular adhesion molecule-1 gene expression in nasal fibroblasts stimulated by IL-1 beta was also suppressed by clarithromycin. Furthermore, electrophoretic mobility shift assay demonstrated that clarithromycin reduced DNA-binding activity of NF-kappa B in both human nasal epithelial cells and fibroblasts stimulated by HIE or IL-1 beta, respectively. CONCLUSION: The present results suggest that clarithromycin may reduce gene expression of proinflammatory cytokines and adhesion molecules from nasal mucosa at the transcriptional factor level and exert an anti-inflammatory effect on nasal mucosa in chronic sinusitis.
Recurrent respiratory papillomatosis (RRP) is highly prevalent in Thailand, with the human papillomavirus (HPV) type-11 being the most widespread. In this study, we isolated the HPV type-11 (HPV-11) genome from subjects with RRP and subcloned the E6 and L2 open reading frames (ORFs) with the expression vectors pEX1 and pEX3. The recombinant E6/beta-gal and L2/beta-gal fusion proteins were expressed in E. coli. Using the recombinant proteins, we demonstrated the presence of antibodies against HPV-11 E6 and L2 in RRP patients by Western blot analysis. The prevalence of seropositivity for HPV-11 E6 and L2 were 5% (1/20) and 10% (2/20), respectively. Although RRP is caused by infection on the mucosal surface, it appears that an immune response occurs against viral proteins expressed in the epithelial lesions.
The problems with the system for reconstructive surgery for head and neck cancers in our department where plastic surgeons are regularly invited from distant facilities were discussed. Especially, the importance of preoperative discussion between the plastic surgeons and ENT doctors was stressed. The computer system with Internet will play an important role in exchanging information such as CT scans and MRI scans between the plastic surgeons and ENT doctors. Moreover, the establishment of a system to prevent postoperative troubles such as closure of anastomosed vessels was stressed. Twenty-seven re-vascularized free flaps had been transferred with a 96.3% success rate (26 of 27 cases). We concluded that the system for reconstructive surgery under the multiple facility cooperation system worked well.
Twelve instances of preoperative autologous blood donation were assessed in 10 patients with malignant tumors of the head and neck region. All patients received preoperative radiation therapy and chemotherapy and most of the cases were also given recombinant human erythropoietin (EPO) and iron sulfate. Hemoglobin concentrations immediately before starting the 800ml blood donations were higher than 13g/dl in 5 cases and less than 13g/dl in 6 cases. The mean hemoglobin concentration just prior to donation and one week after donation in these two groups changed from 14.4g/dl to 12.5g/dl and 11.7g/dl to 11.7g/dl, respectively. A 1000ml blood donation was carried out in only one patient, and the hemoglobin concentrations just before donation and one week after donation were 11.4g/dl and 10.5g/dl, respectively. Only half of the blood volume scheduled, 400ml, was achieved in 3 of the 12 cases attempted because of fever, diarrhea and a change of the day of surgery, respectively. The mean estimated blood loss in the 10 patients that underwent surgery as planned was 898ml, and allogenic blood transfusion was avoided in all cases. We concluded that 800-1000ml preoperative autologous blood donation can be performed safely in patients with advanced malignant tumors of the head and neck region who have undergone preoperative radiation therapy and chemotherapy by giving EPO and iron sulfate.
A 32-year-old female with bilateral congenital stapes fixation accompanied by bilateral proximal symphalangism and bilateral hypermetropia is reported. This is the 19th case of congenital stapes fixation and symphalangism in Japan. Hypermetropia was speculated to be one of the cardinal symptoms of the disease based on the present case and cases previously reported. In this case hypermetropia was due to pure microphthalmos and this was the first report that revealed the cause of hypermetropia in the syndrome. Bilateral stapedotomy were carried out and her hearing loss was improved to the satisfactory level.
Recurrent respiratory papillomatosis (RRP) is highly prevalent in Thailand. In this study, we examined the presence of human papillomavirus (HPV) DNA in 25 RRP patients in Thailand by means of dot blot analysis and/or polymerase chain reaction. Eighty-four percent (21/25) of cases and 4% (1/25) of cases were positive for HPV-11 DNA and HPV-6 DNA, respectively. Three cases (3/25) were negative for all of the examined HPV types. No cases were positive for HPV-16 or 18. Furthermore, we isolated the recombinant HPV-11 DNA clone from a genomic library constructed with the DNA of RRP tissue. The restriction map of the cloned HPV DNA was identical with the map of known HPV-11 DNA. These results suggest at least that no specific HPV type or subtype is likely to be associated with RRP in Thailand.
Papillomaviral E2 genes encode proteins that regulate viral transcription. While the full-length bovine papillomavirus type 1 (BPV-1) E2 peptide is a strong trans activator, the homologous full-length E2 product of human papillomavirus type 16 (HPV-16) appeared to vary in function in previous studies. Here we show that when expressed from comparable constructs, the full-length E2 products of HPV-16 and BPV-1 trans activate a simple E2- and Sp1-dependent promoter up to approximately 100-fold in human keratinocytes and other epithelial cells as well as human and animal fibroblasts. Vaccinia virus-expressed, purified full-length HPV-16 and BPV-1 E2 proteins bound a consensus E2 site with high specific affinities (Kd = approximately 10(-9) M) and stimulated in vitro transcription up to six- to eightfold. In vivo and in vitro trans activation by either E2 protein required cooperation with another activator, such as Sp1, or other factors that interact with papillomavirus promoters, such as AP-1, Oct-1, nuclear factor 1/CTF, transcriptional enhancer factor 1, or USF. The glutamine-rich domain B of Sp1 or the mutually unrelated activation domains of other transcription factors were necessary and sufficient for cooperation with either E2 factor. We conclude that like BPV-1 E2, the HPV-16 E2 protein has the potential to function as a strong activator of viral gene expression in cooperation with cellular transcription factors.
Interleukin-1 beta (IL-1 beta) is a predominant cytokine in retained paranasal sinus fluid of chronic sinusitis where infiltration by polymorphonuclear neutrophils (PMNs) of nasal and paranasal mucosa is characteristic. The authors investigated the localization of IL-1 beta messenger RNA (mRNA) in the maxillary sinus mucosa of patients with chronic sinusitis, using digoxigenin-labeled oligonucleotide probes. IL-1 beta mRNA was detected in some extravascular PMNs and small numbers of mononuclear leukocytes but was not detected in other tissue cells or intravascular leukocytes. The expression and distribution of the cell adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and endothelial-leukocyte adhesion molecule-1 (ELAM-1), were also studied in cultured human mucosal microvascular endothelial cells and in the maxillary sinus mucosa in chronic sinusitis by immunohistochemistry using monoclonal antibodies against these cell adhesion molecules. Only ICAM-1 was expressed on cultured human mucosal microvascular endothelial cells without IL-1 beta stimulation. With IL-1 beta activation of these cells, ELAM-1 was expressed strongly and the expression of ICAM-1 was enhanced. In the maxillary sinus mucosa, ICAM-1 was strongly and universally expressed on endothelial cells of all small vessels, whereas ELAM-1 was expressed only in the subepithelial region. These findings suggest that IL-1 beta, one of mediators in chronic sinusitis, is produced by PMNs, induces the expression of ICAM-1 and ELAM-1 on endothelial cells, and, thereby, stimulates PMN infiltration in chronic sinusitis.
When an HRP or WGA-HRP solution was injected into the rostral midbrain including the oculomotor visceral nuclei, densely distributed HRP/WGA-HRP-positive granules were observed around the ganglion neurons in the accessory ciliary ganglion (ACG) and ectopic neurons in the communicating branch from the long ciliary nerve to the ACG. The same injections labeled fibers within the communicating branch as well as the fibers between the ACG and the main ciliary ganglion (CG). These findings indicate that some oculomotor parasympathetic preganglionic fibers reach the ACG bypassing the CG by way of the trigeminal nerve.
Twenty-eight cases of non-Hodgkin's lymphoma in nasosinus region were studied for tumor-surface marker phenotype, anti-HTLV-I antibody titer and histopathologic correlation with clinical features. The incidence of these cases was 25% of all lymphomas in head and neck region from 1978 to 1987. In 16 cases, 12 (75%) had T-cell markers and 4 (25%) had B-cell markers. Of 11 cases, 6 (54.5%) were anti-HTLV-I antibody carriers. Five year survival ratio of all patients was 4.5%. The survival curve for T-cell lymphoma patients was characterized by a rapid initial decline. The survivals of the group of anti-HTLV-I antibody carriers was estimated inferior to those of non-carriers. Correlation of HTLV-I to prognosis of these patients was discussed.
The relationship between adenoidal-nasopharyngeal ratios (AN ratios) and sinusitis were evaluated in 404 children. The AN ratios of children with snoring, mouth breathing, or nasal obstruction (major symptoms of adenoidal enlargement) were significantly higher than those of children without these symptoms. However, the AN ratios of children with sinusitis were almost equal to those of normal children.