Biomedical subjects

T Fukami

Publications and source records attributed to T Fukami.

At least 19 recordsLinked to original sources

PubMed2006

CYP2A7 polymorphic alleles confound the genotyping of CYP2A6*4A allele.

Human cytochrome P450 (CYP) 2A6 metabolizes nicotine to cotinine. Genetic polymorphisms of CYP2A6 contribute to the interindividual variability of nicotine metabolism. We encountered some subjects possessing two copies of the CYP2A6 gene, although they were genotyped as heterozygotes of the CYP2A6*4A allele (entire CYP2A6 gene deleted allele). From the subjects, we found CYP2A7 polymorphic alleles (CYP2A7*1B, CYP2A7*1C, and CYP2A7*1D) in which the sequences in the 3'-flanking region were converted to the corresponding CYP2A6 sequences, being confused with the CYP2A6*4A. These allele frequencies in European-Americans (n=187) were 1.3, 2.1, 0.3%, respectively, but these were very rare in African-Americans (n=176), Japanese (n=184), and Koreans (n=209). By an improved genotyping method, the allele frequency of CYP2A6*4A of 3.7% in European-Americans was corrected to 0%. The comprehensible and reliable genotyping method developed in this study would be useful to evaluate associations between the genotype and phenotype.

Aryl Hydrocarbon Hydroxylases↗

PubMed2001

Isolation of the TSLL1 and TSLL2 genes, members of the tumor suppressor TSLC1 gene family encoding transmembrane proteins.

We have recently identified the TSLC1 gene as a novel tumor suppressor in human non-small cell lung cancers. TSLC1 encodes a membrane glycoprotein with an extracellular domain homologous to those of immunoglobulin superfamily proteins. Truncation of TSLC1 in the cytoplasmic domain in a primary human tumor suggests that this domain is important for tumor suppressor activity. Here, we report the isolation of two TSLC1-like genes, TSLL1 and TSLL2, based on their structural homology with the sequences corresponding to the cytoplasmic domain of TSLC1. Significant similarity was also observed in the extracellular domain as well as in the overall gene structure, indicating that these three genes form a unique subfamily (the TSLC1-gene family) in the immunoglobulin superfamily genes. In contrast to the ubiquitous expression of TSLC1, TSLL1 is expressed exclusively in adult and fetal human brain, while TSLL2 is expressed in several specific tissues including prostate, brain, kidney and some other organs. Expression of TSLL1 and TSLL2 was lost or markedly reduced in many human glioma cell lines or some prostate cancer cell lines, suggesting that loss of expression of these genes might be involved in some human cancers.

Adult↗

PubMed2001

Identification and characterization of a second melanin-concentrating hormone receptor, MCH-2R.

Melanin-concentrating hormone (MCH) is a 19-aa cyclic neuropeptide originally isolated from chum salmon pituitaries. Besides its effects on the aggregation of melanophores in fish several lines of evidence suggest that in mammals MCH functions as a regulator of energy homeostasis. Recently, several groups reported the identification of an orphan G protein-coupled receptor as a receptor for MCH (MCH-1R). We hereby report the identification of a second human MCH receptor termed MCH-2R, which shares about 38% amino acid identity with MCH-1R. MCH-2R displayed high-affinity MCH binding, resulting in inositol phosphate turnover and release of intracellular calcium in mammalian cells. In contrast to MCH-1R, MCH-2R signaling is not sensitive to pertussis toxin and MCH-2R cannot reduce forskolin-stimulated cAMP production, suggesting an exclusive G(alpha)q coupling of the MCH-2R in cell-based systems. Northern blot and in situ hybridization analysis of human and monkey tissue shows that expression of MCH-2R mRNA is restricted to several regions of the brain, including the arcuate nucleus and the ventral medial hypothalamus, areas implicated in regulation of body weight. In addition, the human MCH-2R gene was mapped to the long arm of chromosome 6 at band 6q16.2-16.3, a region reported to be associated with cytogenetic abnormalities of obese patients. The characterization of a second mammalian G protein-coupled receptor for MCH potentially indicates that the control of energy homeostasis in mammals by the MCH neuropeptide system may be more complex than initially anticipated.

Amino Acid Sequence↗

PubMed2001

Thoracoscopic surgery and conventional open thoracotomy in metastatic lung cancer.

BACKGROUND: We performed a retrospective comparison of the oncological outcome of thoracoscopic surgery for pulmonary metastasis with that of conventional open thoracotomy. METHODS: The patient population for our retrospective comparison was comprised of 45 patients undergoing pulmonary resections via video-assisted thoracoscopy (thoracoscopy group) and 55 undergoing similar resections by open thoracotomy (open group) for pulmonary metastases between 1994 and 1999. RESULTS: Solitary metastasis was resected more frequently with thoracoscopy than open thoracotomy. There were no significant intergroup differences in rates of local recurrence from the initial pulmonary resection site. The actuarial 1-year, 2-year, and 3-year survival rates were, respectively, 82.8%, 70.0%, and 62.3% in the thoracoscopy group and 93.6%, 64.6%, and 52.7% in the open group. The rates of pulmonary recurrence and survival also did not differ significantly between the two groups with solitary metastases. CONCLUSION: Thoracoscopic surgery for metastatic lung disease appears to be feasible as long as the preoperative metastatic tumor evaluation using chest computed tomography (CT) is accurate.

Actuarial Analysis↗

PubMed2001

Elevated expression of interleukin-18 in the granulomatous lesions of muscular sarcoidosis.

In an attempt to understand the role of interleukin-18 (IL-18) in the pathogenesis of sarcoidosis, we examined the expression of IL-18 in normal muscle and in muscle biopsies from six patients with muscular sarcoidosis. Western blot analysis demonstrated that IL-18 was identified only in homogenates of granulomatous muscle tissues, but not in normal muscle tissue homogenates. By immunohistochemistry, strongly IL-18-positive cells were distributed predominantly at the boundary zone of the granulomas. They were recognized as activated macrophages by double staining with anti-CD68. Epithelioid cells showed only faint reactivity. Serum IL-18 levels of patients with sarcoidosis were significantly increased compared to those of healthy volunteers. Unlike protein expression, IL-18 mRNA expression was detected even in normal muscles. Our results coupled with those of previous investigations demonstrating activity of IL-18 in inducing interferon-gamma production suggest a significant role of IL-18 in the pathogenesis of sarcoidosis.

Adult↗

PubMed2001

Effects of lung metastasectomy on respiratory function.

OBJECTIVE: The effects of lung metastasectomy on respiration were evaluated. SUBJECTS: From 1961 to 1999, 203 patients underwent lung metastasectomy. Of these, 102 patients who had undergone only partial lung resection, had undergone sufficient respiratory function testing, and had not suffered from other lung disease were included. RESULTS: Unilateral thoracoscopic surgery caused less decrease in percent vital capacity than surgery through a posterolateral thoracotomy, in both the early (rate of decrease: 16.2 +/- 9.1 vs. 33.0 +/- 12.0%, p < 0.01), and the late postoperative period (2.0 +/- 4.5 vs. 17.8 +/- 6.0%, p < 0.0001). Two thoracoscopic operations caused less decrease in percent vital capacity than 2 operations through a posterolateral thoracotomy (21.3 +/- 13.8 vs. 61.1 +/- 8.1%, p = 0.02). Bilateral metastasectomy through a median sternotomy caused less decrease in percent vital capacity than that through posterolateral thoracotomies (45.5 +/- 13.7 vs. 60.8 +/- 8.8%, p < 0.05). Four (36%) of 11 patients who had undergone 3 or more metastasectomies exhibited dyspnea of degree 3 or higher on the Hugh-Jones classification. CONCLUSIONS: Thoracoscopic metastasectomy and metastasectomy through a median sternotomy caused less restrictive respiratory dysfunction than metastasectomy through a posterolateral thoracotomy. Since metastatic lung disease often necessitates repeated metastasectomy, and repeated metastasectomy often causes severe restrictive respiratory dysfunction, metastasectomy should be performed with a less invasive procedure.

Adult↗

PubMed2001

Tonsillar metastasis from large cell carcinoma of the lung.

A 52-year-old female was referred to our department for treatment of a left lung tumor, 80 mm in diameter, arising in the left S1 + 2. The patient's chief complaint was persistent dry cough and spiking fever. Left upper lobectomy with hilar and mediastinal lymph node dissection (ND2a) was performed, and the pathological diagnosis was primary large cell carcinoma of the lung, p-T3N0M0. At one week after being discharged, the patient visited our outpatient clinic complaining of a sore throat. A tumor in the right tonsil was discovered, and excisional biopsy revealed it to be metastasis from the large cell carcinoma of the lung. Right cervical lymph node metastasis was also detected, and the patient was treated by combined chemo-radiotherapy, resulting in a complete remission.

Carcinoma, Large Cell↗

PubMed2001

Malignant fibrous histiocytoma in the anterior mediastinum.

The origin of a malignant fibrous histiocytoma in the anterior mediastinum is rare. A 71-year-old man was referred to our department for treatment of a tumor 15 cm in diameter originating in the anterior mediastinum. Malignant fibrous histiocytoma was suspected after a percutaneous needle biopsy. Systemic screening showed no other organ involvement, and the tumor was considered to be a primary mediastinal tumor. The tumor, resected through a median sternotomy, was well encapsulated and resection was complete. The pathological diagnosis was malignant fibrous histiocytoma originating in the anterior mediastinum.

Aged↗

PubMed2001

Different binding sites for the neuropeptide Y Y1 antagonists 1229U91 and J-104870 on human Y1 receptors.

The peptidic Y1 antagonist 1229U91 and the non-peptidic antagonist J-104870 have high binding affinities for the human Y1 receptor. These Y1 antagonists show anorexigenic effects on NPY-induced feeding in rats, although they have completely different structures and molecular sizes. To identify the binding sites of these ligands, we substituted amino acid residues of the human Y1 receptor with alanine and examined the abilities of the mutant receptors to bind the radio-labeled ligands. Alanine substitutions, F98A, D104A, T125A, D200A, D205A, L215A, Q219A, L279A, F282A, F286A, W288A and H298A, in the human Y1 receptor lost their affinity for the peptide agonist PYY, but not for 1229U91 and J-104870, while L303A and F173A lost affinity for 1229U91 and J-104870, respectively. N283A retained its affinity for 1229U91, but not for PYY and J-104870. Y47A and N299A retained their affinity for J-104870, but not for PYY and 1229U91. W163A and D287A showed no affinity for any of the three ligands. Taken together, these data indicate that the binding sites of 1229U91 are widely located in the shallow region of the transmembrane (TM) domain of the receptor, especially TM1, TM6 and TM7. In contrast, J-104870 recognized the pocket formed by TM4, TM5 and TM6, based on the molecular modeling of the Y1 receptor and J-104870 complex. In conclusion, 1229U91 and J-104870 have high affinities for Y1 receptors using basically different binding sites. D287 of the common binding site in the TM6 domain could be crucial for the binding of Y1 antagonists.

Alanine↗

PubMed2001

A typical Y1 receptor regulates feeding behaviors: effects of a potent and selective Y1 antagonist, J-115814.

Neuropeptide Y (NPY) is a potent feeding stimulant. The orexigenic effect of NPY might be caused in part by the action of Y1 receptors. However, the existence of multiple NPY receptors including a possible novel feeding receptor has made it difficult to determine the relative importance of the Y1 receptor in feeding regulation. Herein we certified that the Y1 receptor is a major feeding receptor of NPY by using the potent and selective Y1 antagonist (-)-2-[1-(3-chloro-5-isopropyloxycarbonylaminophenyl)ethylamino]-6-[2-(5-ethyl-4-methyl-1,3-thiazol-2-yl)ethyl]-4-morpholinopyridine (J-115814) and Y1 receptor-deficient (Y1-/-) mice. J-115814 displaced (125)I-peptide YY binding to cell membranes expressing cloned human, rat, and murine Y(1) receptors with K(i) values of 1.4, 1.8, and 1.9 nM, respectively, and inhibited NPY (10 nM)-induced increases in intracellular calcium levels via human Y1 receptors (IC(50) = 6.8 nM). In contrast, J-115814 showed low affinities for human Y2 (K(i) > 10 microM), Y4 (K(i) = 640 nM) and Y5 receptors (K(i) = 6000 nM). Intracerebroventricular (ICV) (10-100 microg) and intravenous (IV) (0.3-30 mg/kg) administration of J-115814 significantly and dose-dependently suppressed feeding induced by ICV NPY (5 microg) in satiated Sprague-Dawley rats. Intraperitoneal (IP) administration of J-115814 (3-30 mg/kg) significantly attenuated spontaneous feeding in db/db and C57BL6 mice. Feeding induced by ICV NPY (5 microg) was unaffected by IP-injected J-115814 (30 mg/kg) in Y1-/- mice and was suppressed in wild-type and Y5-/- mice. These findings clearly suggest that J-115814 inhibits feeding behaviors through the inhibition of the typical Y1 receptor. We conclude that the Y1 receptor plays a key role in regulating food intake.

Animals↗

PubMed2000

L-152,804: orally active and selective neuropeptide Y Y5 receptor antagonist.

Neuropeptide Y (NPY) elicits food intake through the action of hypothalamic G-protein-coupled receptors. Previous publications indicate that the Y5 receptor may represent one of these postulated hypothalamic "feeding" receptors. Using a potent and orally available Y5 antagonist L-152,804, we evaluated the involvement of the Y5 receptor in feeding regulation. L-152,804 displaced [125I]peptide YY (PYY) binding to human and rat Y5 receptors with Ki values of 26 and 31 nM, respectively, and inhibited NPY (100 nM)-induced increase in intracellular calcium levels via human Y5 receptors (IC50 = 210 nM). L-152,804 did not show significant affinity for human Y1, Y2, and Y4 receptors at a dose of 10 microM. Intracerebroventricular (i.c.v.) (30 microg) or oral (10 mg/kg) administration of L-152,804 significantly inhibited food intake evoked by i.c.v.-injected bovine pancreatic peptide (bPP, 5 microg; a moderately selective Y4, Y5 agonist) in satiated SD rats. However L-152,804 did not significantly inhibit i.c.v. NPY (5 microg; a Y1, Y2, Y5 agonist)-induced food intake. These findings suggest that L-152,804 is a selective and potent non-peptide Y5 antagonist with oral bioavailability and brain penetrability. In addition, the anorexigenic effects of L-152,804 on bPP-induced feeding revealed participation of the Y5 receptor in feeding regulation, while i.c.v. administration of NPY does not appear to significantly contribute to Y5 stimulated food intake. We conclude that the potent and orally active Y5 antagonist, L-152,804, represents a useful tool to address the physiological role of the Y5 receptor.

Administration, Oral↗

PubMed2000

Fluorometric assay of DNA binding proteins using class IIS endonuclease.

Fluorometric DNA-protein binding assay was developed with class IIS restriction enzyme and FITC-labeled DNA fixed on magnetic beads. We used this assay with AP1 and FokI as a case study to quantify DNA-protein binding. Fluorescent images of the treated beads showed that the transcriptional factor bound to its DNA site and inhibited the restricted digestion effectively. The affinity of AP1 to its site was estimated as a Kd value, which was approximately 10 nM.

DNA-Binding Proteins↗

PubMed2000

A novel method for DNA molecular counting.

We have developed a new method for counting DNA molecules using 'capillary-plates' consisting of a large number of small glass-capillary 'channels' fused together in parallel. PCR mixtures containing serially diluted DNA templates with the DNA indicator dye Hoechst 33258 were poured into the plates and sealed with silicone rubber-plates. Following 40 PCR cycles, fluorescence microscopy revealed that the fluorescence in some channels had increased about three-times more than in others at template concentrations of 1 fM or lower. No bright fluorescence was observed in the absence of template. The relationship between the proportion of fluorescent channels in the capillary-plates and the template concentrations was linear according to Poisson probabilities in the range of 0.1-1,000 aM. These results demonstrate the amplification of single templates in the channels, and that a small number of templates could be quantified by counting the proportion of positive channels on the capillary-plates.

Bisbenzimidazole↗

PubMed2000

Role of the Y1 receptor in the regulation of neuropeptide Y-mediated feeding: comparison of wild-type, Y1 receptor-deficient, and Y5 receptor-deficient mice.

Neuropeptide Y (NPY) increases food intake through the action of hypothalamic NPY receptors. At least six subtypes of NPY, peptide YY (PYY), and pancreatic polypeptide (PP) receptors have been identified in mice. Although the involvement of Y1 and Y5 receptors in feeding regulation has been suggested, the relative importance of each of these NPY receptors and the participation of a novel feeding receptor are still unclear. To address this issue, we generated a Y1 receptor-deficient (Y1-/-) and a Y5 receptor-deficient (Y5-/-) mouse line in which we directly compared the orexigenic effects of NPY and its analogs after intracerebroventricular (icv) administration. The icv NPY-induced food intake was remarkably reduced in Y1-/- mice, but was not significantly altered by inactivation of the Y5 receptor. The Y1 receptor therefore plays a dominant role in NPY-induced feeding. Stimulation of feeding by moderately selective Y5 agonists [PYY-(3-36), human PP, and bovine PP] was reduced in Y5-/- mice, although food intake did not decrease to vehicle control levels. These results indicate that the Y5 receptor functions as one of the feeding receptors. In addition, the finding that Y5-preferring agonists still induce food intake in Y5-/- mice suggests a role for another NPY receptor(s), including the possibility of novel NPY receptors. Surprisingly, despite the limited efficacy of PYY-(3-36) and PPs at the Y1 receptor, food consumption induced by these agonists was significantly diminished in Y1-/- mice compared with that in wild-type controls. These observations suggest that the feeding stimulation induced by NPY and its analogs may be directly or indirectly modulated by the action of the Y1 receptor. We conclude that multiple NPY receptors, possibly including the novel feeding receptor, are involved in the feeding response evoked by NPY and its analogs. Among them, the Y1 receptor plays a key role in NPY-induced feeding in mice.

Animals↗

PubMed2000

A case of sarcoidosis presenting as multiple pulmonary nodules, nasopharyngeal and cerebellopontine tumors.

A 64-year-old woman presented with multiple pulmonary nodules, and after spontaneous regression of the pulmonary lesions in six months, nasopharyngeal and right cerebellopontine tumors developed. Noncaseous epithelioid cell granulomas were demonstrated histologically in both resected pulmonary and nasopharyngeal tumors. She complained of hearing loss and tinnitus probably due to the cerebellopontine tumors. Corticosteroid therapy resulted in the improvement of these symptoms and the gradual decrease of tumor size. Although histological probe of the cerebellopontine tumors was not diagnostic, this patient was finally diagnosed as having sarcoidosis, based on the clinicopathological features, including systemic granulomatous lesions, MRI findings, and good response to corticosteroid therapy. The diagnosis of sarcoidosis is sometimes difficult when its clinical manifestations are uncommon.

Cerebellar Diseases↗

PubMed2000

[Pulmonary eosinophilic granuloma associated with recurrent pneumothorax: report of a case].

A 25-years-old male, current smoker, was referred to our department for treatment of the right pneumothorax due to pulmonary eosinophilic granuloma. He underwent thoracoscopic partial resection of the right lung. Immunostaining of the specimen revealed a lot of Langerhans cells positive for S 100 protein, suggesting active lesion. After cessation of smoking, no recurrence of pneumothorax or progression of disease have been observed for 13 months.

Adult↗

PubMed1999

The novel neuropeptide Y Y(1) receptor antagonist J-104870: a potent feeding suppressant with oral bioavailability.

Neuropeptide Y (NPY) is known to induce robust feeding through the action of NPY receptors in the hypothalamus. Among the subtypes of NPY receptors, Y(1) receptors may play a key role in feeding regulation. In the present study, we demonstrated that a novel Y(1) antagonist, J-104870, shows high selectivity and potency for the Y(1) receptor with an anorexigenic effect on NPY-mediated feeding. J-104870 displaced [(125)I]peptide YY (PYY) binding to cloned human and rat Y(1) receptors with K(i) values of 0.29 and 0.54 nM, respectively, and inhibited the NPY (10 nM)-induced increase in intracellular calcium levels (IC(50) = 3.2 nM) in cells expressing human Y(1) receptors. In contrast, J-104870 showed low affinities for human Y(2) (K(i) > 10 microM), Y(4) (K(i) > 10 microM), and Y(5) receptors (K(i) = 6 microM). In rat hypothalamic membranes, J-104870 also completely displaced the binding of [(125)I]1229U91, which is known to bind to the typical Y(1) receptor, with a high affinity (K(i) = 2.0 nM). Intracerebroventricular (ICV) injection of J-104870 (200 microg) significantly suppressed NPY (5 microg)-induced feeding in satiated Sprague-Dawley rats by 74%. Furthermore, ICV and oral administration of J-104870 (200 microg and 100 mg/kg, respectively) significantly suppressed spontaneous food intake in Zucker fatty rats. These findings suggested that J-104870 is a selective and potent nonpeptide Y(1) antagonist with oral bioavailability and brain penetrability. In addition, the anorexigenic effect of J-104870 clearly revealed the participation of the Y(1) receptor in NPY-mediated feeding regulation. The potent and orally active Y(1) antagonist J-104970 is a useful tool for elucidating the physiological roles of NPY in obesity.

Administration, Oral↗

PubMed1999

Interleukin-4-producing cells in idiopathic pulmonary fibrosis: an immunohistochemical study.

OBJECTIVE: Interleukin-4 stimulates collagen synthesis in fibroblasts in vitro. The purpose of this study was to clarify the role of interleukin-4 in the pathogenesis of idiopathic pulmonary fibrosis. METHODOLOGY: We compared interleukin-4 expressing cells in tissues from 11 patients with pathologically confirmed idiopathic pulmonary fibrosis with early stage inflammatory lesions versus advanced lesions. We also performed double staining in order to elucidate which inflammatory cells produce interleukin-4 in idiopathic pulmonary fibrosis. RESULTS: Greater numbers of interleukin-4 positive cells were observed in the advanced lesions than in the early lesions. We also found that early stage lesions showed a predominance of CD3/interleukin-4 double positive cells whereas, the advanced stage lesions had higher percentages of the EG2/interleukin-4 double positive cells and the tryptase/interleukin-4 double positive cells. CONCLUSIONS: Our results suggest that interleukin-4 may be involved in progression of interstitial fibrosis in idiopathic pulmonary fibrosis, in which we confirmed a significant contribution of eosinophils and mast cells to interleukin-4 production.

Adult↗