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Biomedical subjects

K Asano

Publications and source records attributed to K Asano.

At least 19 recordsLinked to original sources

Gene expressions of canine angiopoietin-1 and -2 in normal tissues and spontaneous tumours.

The angiopoietin (Ang) family of proteins are central to the regulation of angiogenesis. The purposes of this study were to determine cDNA sequences of canine Ang-1 and Ang-2 and investigate their expressions in normal tissues and spontaneous tumours. The cDNA sequences of canine Ang-1 and Ang-2 were 1,494 and 1,488 bp, and the deduced amino acid sequences were 497 and 495 residues, respectively. The cDNA sequences of canine Ang-1 and Ang-2 showed high homology with those of the other mammalian species. Canine Ang-1 and Ang-2 mRNA were detectable in all 22 normal tissues and spontaneous tumours. Higher mRNA expression level of canine Ang-2 was demonstrated in mammary simple carcinomas, haemangiosarcoma and hepatocellular carcinoma in comparison with normal tissues.

Amino Acid Sequence↗

Nitric oxide donor-induced inhibition of pregnant rat uterine spontaneous contractile activity and release of nitric oxide from uterus measured by microdialysis.

Our aim was to study whether nitric oxide (NO) donor-induced inhibition of pregnant rat myometrium contractility correlates with the release of NO. Uterine rings from mid-pregnant and late pregnant Sprague-Dawley rats were used for isometric tension recording. Concentration-response relationships to sodium nitroprusside (SNP), nitroglycerine (NTG) and diethylamine (DEA)/NO were assessed. The time course of NO release after addition to the organ chambers of the 3 NO-donors was assessed by the detection of NO products NOx (NO3+NO2) using the microdialysis probe by a HPLC-NO detector system. DEA/ NO induced greater inhibition of the spontaneous contractile activity of uterine rings from mid-pregnant rats than SNP or NTG. In uterine rings from late pregnant rats, however, the maximal inhibition of the contractility by all 3 NO-donors were significantly less. The NOx levels measured in the uterine ring walls from either mid-pregnant or late pregnant rats significantly increased after DEA/ NO as compared to the basal levels or the levels after NTG or SNP. The decrease of NO-donor-induced inhibition of rat myometrium contractility, with unchanged formation of NOx, at term, suggests that the changes in NO signaling are responsible for gestational age-dependent attenuation of the inhibitory effect.

Animals↗

Effects of sex steroids on expression of adenylyl cyclase messenger RNA in rat uterus.

The aim of our study was to investigate the effects of estradiol (E2) and/or progesterone (P4) bilateral ovariectomized rats on gene expressions of adenylyl cyclases (AC2, AC4, AC6, AC9). Non-pregnant (NP) Wister rats (between post-natal 10 and 12 weeks) and normal pregnant rats (between postnatal 10 and 14 weeks) were used. NP rats were divided into five groups: 1) control NP rats, 2) ovariectomy rats (OVX), 3) OVX injected with E2 (6 microg) for 4 days (E2), 4) OVX injected with P4 (5 mg) for 4 days (P4), 5) ovariectomy rats injected with E2 (6 microg) and P4 (5 mg) for 4 days (E2/P4). The ovariectomized rats were sacrificed by anesthesia of diethyl ether after 24 h from the last injection. The pregnant group was sacrificed in the same way on 14 (P14), 17 (P17) and 21 (P21) of pregnancy, because the days between P14 and P17 were the second trimester of pregnancy and P21 was the third trimester. Total RNA were prepared from the uteri of each subject, and expression of ACs was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Expression of mRNAs for AC2, AC4, AC6 and AC9 were maximally increased on P17 and then decreased on P21. In OVX rats, expression of mRNAs for AC2, AC4, and AC9 were decreased as compared to NP. AC2 mRNA was significantly increased by E2 treatment. AC4 and AC9 mRNA was significantly increased by injection of E2 or P4. AC6 mRNA level was not affected by E2 and/or P4. In conclusion, AC2, AC4 and AC9 gene expression is affected by estrogen and/or P4. On the contrary, AC6 gene expression was not changed by OVX or hormone treatment. Since, all ACs mRNA examined here are increased during pregnancy, AC6 may be affected by other factor(s) other than estrogen and P4.

Adenylyl Cyclases↗

Pulmonary adenosquamous carcinoma in a dog.

A mass that developed in the lung of a 10-year-old mixed-breed dog was pathologically examined. Histopathological examination showed papillary and tubular growth of glandular epithelium-like cells in some areas and growth of squamous cells arranged in nests in other areas, showing coexistence of adenocarcinoma and squamous cell carcinoma in a lung tumour. Immunohistochemical staining with anti-keratin-cytokeratin antibody was strongly positive for cytoplasms in both components. Electron microscopically, the neoplastic cells of the adenocarcinoma component had features of glandular cells, with microvilli, numerous free ribosomes, large round secretory granules and intercellular desmosomes. Non-keratinized squamous cells had tonofilaments and intercellular desmosomes. These findings led to the diagnosis of primary adenosquamous carcinoma, which demonstrates phenotypic profiles characteristic of both epidermal keratinocytes and glandular epithelium.

Animals↗

[Surgical resection for T4 lung cancer invading thoracic aorta].

The prognosis of the lung cancer patients with aortic invasion is thought to be very poor in general. Thoracic aorta resection and reconstruction was performed in 6 patients, aortic arch in 2, descending aorta in 4. An intraoperative and a postoperative major complication occurred in each 1 patient. Five patients survived more than 1 year after operation, and 1 of them has been living without relapse for more than 5 years. Pulmonary resection with the involved aorta can be done safely using cardiopulmonary bypass, with encouraging long-term survivals in patients without N2 or N3 nodal metastasis.

Aged↗

Plasma eotaxin level and severity of asthma treated with corticosteroid.

Our understanding of asthma severity was advanced by the identification of biomarkers which account for differences in lung function impairment. We tried to examine the effects of corticosteroid treatment on known correlates of asthma severity including peripheral eosinophil counts, total IgE, IL-5, and eotaxin Levels in plasma. We compared these biomarkers among groups of stable asthmatics categorized by the dose of corticosteroid (N: steroid-free, n = 25; L: low-dose inhaled, n = 27; MH: medium or high-dose inhaled, n= 19; O: inhaled plus oral, n= 8). Next we compared these markers and peak expiratory flow rate (PEFR) in unstable asthmatics before and after treatment with steroids (n = 22). Eotaxin levels in the O group were higher than those in the N and MH groups (P < 0.05). Logistic regression analysis demonstrated that plasma eotaxin level was correlated with the severity of asthma defined by treatment intensity (P = 0.01) and % FEV1 (P = 0.04) while the other markers were not. Eotaxin levels did not change after steroid treatment in unstable patients, whereas eosinophil counts decreased in parallel with PEFR. Among biomarkers of asthma severity studied, plasma eotaxin level was not significantly affected by corticosteroid treatment, and was associated with the severity even in the presence of steroid therapy.

Administration, Inhalation↗

Relationship between serum testosterone and activities of testicular enzymes after continuous and intermittent training in male rats.

To gain more information on the effects of training types on testosterone secretion, the present study investigated the relationship between serum testosterone (ST) and the activities of oxydoreductive enzymes in Leydig cells to continuous and intermittent training regimes. Male rats swam with a load of 3.5 % body weight for 90 min in the continuous training group, and 15 min separated by a 7-min rest interval x 6 times in the intermittent training group, 6 days per week for 5 weeks. ST were measured immediately and 24 h after exercise, and the activities of SDH, LDH and G6PDH in Leydig cells were measured 24 h after exercise, following 5 weeks of training. It was found that ST declined following continuous (0.54 +/- 0.32 nmol/l) and intermittent (1.64 +/- 1.80 nmol/l) exercise compared to sedentary group (9.55 +/- 5.17 nmol/l). This diminishing effect on ST was still significant 24 h after continuous exercise (5.96 +/- 2.79 nmol/l), not after intermittent exercise (7.41 +/- 4.77 nmol/l). The activities of SDH and LDH increased, whereas G6PDH decreased in Leydig cells, after both continuous and intermittent training. SDH and G6PDH showed the high activities in the intermittent training group relative to continuous training group. These differences in the activities of SDH and G6PDH might be considered as the possible causes for ST responses to training types.

Animals↗

Expression of inducible nitric oxide synthase messenger RNA, but not guanylate cyclase messenger RNA, depends on gestational age in rat myometrium.

We studied the expression of inducible nitric oxide (NO) synthase (iNOS) and soluble guanylate cyclase (sGC) mRNAs in pregnant rat myometrium. Expression of iNOS and sGC alpha1, beta1 and beta2 mRNA was analyzed in non-pregnant and pregnant (days 10, 14, 17 and 21) Wistar rats by reverse transcription-polymerase chain reaction. Expression of iNOS mRNA increased during pregnancy but decreased on day 21 of gestation. Expression of GC alpha1 mRNA was greater than GC beta1 mRNA at all time points. Expression of uterine GC alpha1 and GC beta1 mRNA did not change significantly during pregnancy and did not differ significantly from non-pregnant levels. The values of sGC beta2 mRNA were below the limit of detection. In conclusion, the expression of iNOS mRNA increased during pregnancy in the myometrium and decreased at term, while the expression of sGC mRNA was not affected by pregnancy. Thus, it is the changes in NO production, rather than changes in its target, that are responsible for uterine quiescence during pregnancy and initiation of labor.

Animals↗

Suppressive activity of fexofenadine hydrochloride on metalloproteinase production from nasal fibroblasts in vitro.

BACKGROUND: Allergic rhinitis (AR) is an inflammatory disease characterized by nasal wall remodelling with intense infiltration of eosinophils and mast cells/basophils. Matrix metalloproteinases (MMPs), MMP-2 and MMP-9, are the major proteolytic enzymes that induce airway remodelling. These enzymes are also important in the migration of inflammatory cells through basement membrane components. OBJECTIVE: We evaluated whether fexofenadine hydrochloride (FEX), the carboxylic acid metabolite of terfenadine with selective H(1)-receptor antagonist activity, could inhibit MMP production from nasal fibroblasts (NFs) in response to TNF-alpha stimulation in vitro. METHODS: NFs were established from nasal polyp-derived fibroblasts (PFs) taken from patients with AR. Nasal mucosal fibroblasts (MFs) were also induced from nasal mucosal tissues from septal deformity patients without allergy. PF and MF (2 x 10(5) cells/mL, each) were stimulated with TNF-alpha in the presence of various concentrations of FEX. After 24 h, culture supernatants were obtained and assayed for MMP-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 levels by ELISA. The influence of FEX on mRNA expression of MMPs and TIMPs in 4 h-cultured cells was also evaluated by real-time RT-PCR. Furthermore, nuclear factor-kappa B (NF-kappa B) activation in fibroblasts treated with FEX for 4 h was examined by ELISA. RESULTS: FEX at more than 350 ng/mL inhibited the production of MMP-2 and MMP-9 from both PF and MF in response to TNF-alpha stimulation, whereas TIMP-1 and TIMP-2 production was scarcely affected by FEX. FEX also inhibited MMP mRNA expression and NF-kappa B activation in PF and MF after TNF-alpha stimulation. CONCLUSION: The present data suggest that the attenuating effect of FEX on MMP-2 and -9 production from NFs induced by inflammatory stimulation may underlie the therapeutic mode of action of the agent on allergic diseases, including AR.

Adult↗

Optimal degradation rate for collagen chambers used for regeneration of peripheral nerves over long gaps.

The experimental study of peripheral nerve regeneration has depended heavily on the use of a nerve chamber in which the stumps of the transected nerve are inserted. A large variety of chamber fillings and chamber types have been used in an effort to induce a higher quality of regeneration across the gap initially separating the two stumps. In this study we studied the morphology of nerves regenerated across a 15 mm gap following implantation of a series of five chambers. The chambers were fabricated from type I collagen and possessed identical pore volume fractions as well as average pore diameters, but differed in cross-link density continuously along the series. The residual mass of the implanted chambers at 9 weeks was observed to increase continuously with increasing cross-link density along the series, indicating a continuous decrease in degradation rate. The quality of regenerated nerves, determined by the number of large diameter fibers (A-fibers) per nerve, the average diameter of all axons and the ratio of area occupied by axons (N-Ratio), was superior at an intermediate level of chamber degradation rate. The maximal quality of peripheral nerve regeneration corresponded to an optimal degradation rate with an estimated chamber half-life of approximately 2-3 weeks following implantation. A speculative mechanistic explanation of the observed optimum focuses on the hypothetical role of cell and cytokine traffic that may take place through holes in the chamber generated by the degradation process. The data show the presence of a hitherto unreported optimal chamber degradation rate that leads to regenerated nerves of maximum quality.

Absorbable Implants↗

Suppression of matrix metalloproteinase production from nasal fibroblasts by macrolide antibiotics in vitro.

It is well known that low-dose and long-term administration of macrolide antibiotics favourably modify the clinical status of chronic airway inflammatory diseases. However, the therapeutic mode of action of macrolide antibiotics is not well understood. The present study aimed to examine the influence of macrolide antibiotics, roxithromycin (RXM) and josamycin (JM) on matrix metalloproteinase (MMP) production from nasal polyp fibroblasts (NPF) in vitro. NPF, at a concentration of 2.5 x 10(5) cells x mL(-1), were stimulated with tumour necrosis factor (TNF)-alpha in the presence of various concentrations of RXM or JM for 24 h. MMP-2 and -9 levels in culture supernatants were analysed by ELISA, and MMP mRNA expression was examined by RT-PCR. The influence of RXM on nuclear factor (NF)-kappaB and activator protein (AP)-1 activation was also examined. Addition of RXM (but not JM) at 5.0 and 7.5 microg x mL(-1) significantly suppressed the production of MMP-2 and -9 from NPF induced by TNF-alpha stimulation. RXM also suppressed MMP mRNA expression through the inhibition of NF-kappaB and AP-1 activation. The present results suggest that the suppressive activity of roxithromycin on MMP-2 and -9 production is, in part, responsible for the therapeutic action of macrolides on chronic airway inflammatory diseases.

Adult↗

[Extrapleural pneumonectomy after hyperthermo-chemotherapy for the lung cancer patients with malignant pleural effusion].

The prognosis of the lung cancer patients with malignant pleural effusion is extremely poor. Intrapleural perfusion hyperthermo-chemotherapy for the treatment of malignant pleural effusion was performed in 120 lung cancer patients and showed better prognosis than conventional treatment. After successful local control by hyperthermo-chemotherapy, extrapleural pneumonectomy was performed in 8 patients with clinical N 0 or N 1 and M 0 lung cancer. All patients survived for more than 1 year. The only 1 patient survived for more than 3 years and 3 months without recurrence. Almost all patients who died within 2 years had pathological n 2, carcinomatous pericarditis, cancer invasion beyond extrapleural space and so on. Extrapleural pneumonectomy after hyperthermo-chemotherapy can be a choice of surgical treatment if the n 2 disease, pericarditis, and extrapleural invasion of disseminated cancer cells should be excluded precisely.

Adenocarcinoma↗

The incidence of thanatophoric dysplasia mutations in FGFR3 gene is higher in low-grade or superficial bladder carcinomas.

BACKGROUND: The point mutations of fibroblast growth factor receptor 3 (FGFR3) are associated with autosomal dominant human skeletal disorders such as thanatophoric dysplasia (TD). However, point mutations were reported in a small series of bladder carcinomas, suggesting their oncogenic role. In view of these findings, the authors investigated the incidence of TD mutations in the FGFR3 gene in a large series of bladder carcinomas to clarify their role in the progression of bladder carcinoma. METHODS: Specimens of transitional cell carcinoma of the urinary bladder from 81 patients were screened for the FGFR3 mutations (codons 248, 249, 372, 373, 375, 652, 809) that have been reported in TD, using polymerase chain reaction-restriction fragment length polymorphism, single-strand conformation polymorphism, and DNA sequencing. RESULTS: Point mutations were detected in 25 of 81 carcinomas (2 at codon 248, 11 at codon 249, 1 at codon 372, 9 at codon 375, 2 at codon 652). Although no significant relation was found between the occurrence of TD mutations and patient age and clinical status, the incidence of TD mutations was significantly higher in low-grade or superficial tumors than high-grade or muscle invasive tumors. CONCLUSIONS: These findings indicate that TD mutations in the FGFR3 gene do not cause disease progression of bladder carcinoma.

Adult↗

Single nucleotide polymorphism of human platelet-activating factor receptor impairs G-protein activation.

Various proinflammatory and vasoactive actions of platelet-activating factor (PAF) are mediated through a specific G-protein-coupled PAF receptor (PAFR). We identified a novel DNA variant in the human PAFR gene, which substitutes an aspartic acid for an alanine residue at position 224 (A224D) in the putative third cytoplasmic loop. This mutation was observed in a Japanese population at an allele frequency of 7.8%. To delineate the functional consequences of this structural alteration, Chinese hamster ovary cells were stably transfected with constructs encoding either wild-type or A224D mutated PAFR. No significant difference was observed in the expression level of the receptor or the affinity to PAF or to an antagonist, WEB2086, between the cells transfected with wild-type and mutant PAFR. Chinese hamster ovary cells expressing A224D mutant PAFR displayed partial but significant reduction of PAF-induced intracellular signals such as calcium mobilization, inositol phosphate production, inhibition of adenylyl cyclase, and chemotaxis. These findings suggest that this variant receptor produced by a naturally occurring mutation exhibits impaired coupling to G-proteins and may be a basis for interindividual variation in PAF-related physiological responses, disease predisposition or phenotypes, and drug responsiveness.

Adenylate Cyclase Toxin↗

Enhancement of endogenous corticosterone levels by a macrolide antibiotic, roxithromycin in mice.

The influence of roxithromycin (RXM), a macrolide antibiotic, on endogenous corticosterone (CS) levels was examined in BALB/c mice. Mice were sensitized intraperitoneally with two doses of Keyhole Limpet Hemocyanin at 1 week intervals. Mice were given orally 2.5 mg/kg RXM once a day for 14 days starting 7 days after the first sensitization. RXM administration caused markedly increase in endogenous plasma CS levels which was peaked at 60 min after the administration. However, josamycin did not influence on endogenous CS levels in plasma. Injection of dexamethasone inhibits the plasma CS hyperproduction induced by RXM treatment.

Allergens↗