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Biomedical subjects

S Murakami

Publications and source records attributed to S Murakami.

At least 109 records · Page 6Linked to original sources
PubMed2001

Rat mannose-binding protein a binds CD14.

Lipopolysaccharide (LPS) has been known to induce inflammation by interacting with CD14, which serves as a receptor for LPS. Mannose-binding protein (MBP) belongs to the collectin subgroup of the C-type lectin superfamily, along with surfactant proteins SP-A and SP-D. We have recently demonstrated that SP-A modulates LPS-induced cellular responses by interaction with CD14 (H. Sano, H. Sohma, T. Muta, S. Nomura, D. R. Voelker, and Y. Kuroki, J. Immunol. 163:387-395, 2000) and that SP-D also interacts with CD14 (H. Sano, H. Chiba, D. Iwaki, H. Sohma, D. R. Voelker, and Y. Kuroki, J. Biol. Chem. 275:22442-22451, 2000). In this study, we examined whether MBP, a collectin highly homologous to SP-A and SP-D, could bind CD14. Recombinant rat MBP-A bound recombinant human soluble CD14 in a concentration-dependent manner. Its binding was not inhibited in the presence of excess mannose or EDTA. MBP-A bound deglycosylated CD14 treated with N-glycosidase F, neuraminidase, and O-glycosidase, indicating that MBP-A interacts with the peptide portion of CD14. Since LPS was also a ligand for the collectins, we compared the characteristics of binding of MBP-A to LPS with those of binding to CD14. MBP-A bound to lipid A from Salmonella enterica serovar Minnesota and rough LPS (S. enterica serovar Minnesota Re595 and Escherichia coli J5, Rc), but not to smooth LPS (E. coli O26:B6 and O111:B4). Unlike CD14 binding, EDTA and excess mannose attenuated the binding of MBP-A to rough LPS. From these results, we conclude that CD14 is a novel ligand for MBP-A and that MBP-A utilizes a different mechanism for CD14 recognition from that for LPS.

Animals↗
PubMed2001

Overexpression of SH2-containing inositol phosphatase 2 results in negative regulation of insulin-induced metabolic actions in 3T3-L1 adipocytes via its 5'-phosphatase catalytic activity.

Phosphatidylinositol (PI) 3-kinase plays an important role in various metabolic actions of insulin including glucose uptake and glycogen synthesis. Although PI 3-kinase primarily functions as a lipid kinase which preferentially phosphorylates the D-3 position of phospholipids, the effect of hydrolysis of the key PI 3-kinase product PI 3,4,5-triphosphate [PI(3,4,5)P3] on these biological responses is unknown. We recently cloned rat SH2-containing inositol phosphatase 2 (SHIP2) cDNA which possesses the 5'-phosphatase activity to hydrolyze PI(3,4,5)P3 to PI 3,4-bisphosphate [PI(3,4)P2] and which is mainly expressed in the target tissues of insulin. To study the role of SHIP2 in insulin signaling, wild-type SHIP2 (WT-SHIP2) and 5'-phosphatase-defective SHIP2 (Delta IP-SHIP2) were overexpressed in 3T3-L1 adipocytes by means of adenovirus-mediated gene transfer. Early events of insulin signaling including insulin-induced tyrosine phosphorylation of the insulin receptor beta subunit and IRS-1, IRS-1 association with the p85 subunit, and PI 3-kinase activity were not affected by expression of either WT-SHIP2 or Delta IP-SHIP2. Because WT-SHIP2 possesses the 5'-phosphatase catalytic region, its overexpression marked by decreased insulin-induced PI(3,4,5)P3 production, as expected. In contrast, the amount of PI(3,4,5)P3 was increased by the expression of Delta IP-SHIP2, indicating that Delta IP-SHIP2 functions in a dominant-negative manner in 3T3-L1 adipocytes. Both PI(3,4,5)P3 and PI(3,4)P2 were known to possibly activate downstream targets Akt and protein kinase C lambda in vitro. Importantly, expression of WT-SHIP2 inhibited insulin-induced activation of Akt and protein kinase C lambda, whereas these activations were increased by expression of Delta IP-SHIP2 in vivo. Consistent with the regulation of downstream molecules of PI 3-kinase, insulin-induced 2-deoxyglucose uptake and Glut4 translocation were decreased by expression of WT-SHIP2 and increased by expression of Delta IP-SHIP2. In addition, insulin-induced phosphorylation of GSK-3beta and activation of PP1 followed by activation of glycogen synthase and glycogen synthesis were decreased by expression of WT-SHIP2 and increased by the expression of Delta IP-SHIP2. These results indicate that SHIP2 negatively regulates metabolic signaling of insulin via the 5'-phosphatase activity and that PI(3,4,5)P3 rather than PI(3,4)P2 is important for in vivo regulation of insulin-induced activation of downstream molecules of PI 3-kinase leading to glucose uptake and glycogen synthesis.

3T3 Cells↗
PubMed2001

Periodontal regeneration by FGF-2 (bFGF) in primate models.

We recently demonstrated that a topical application of basic fibroblast growth factor (FGF-2; bFGF) to alveolar bone defects in beagle dogs enhanced periodontal regeneration. The purpose of this study was further characterization of the biological effects of FGF-2 in non-human primates. Thirty-two inflamed furcation class II defects were surgically created in 4 male primates. The gelatinous carrier alone or the carrier containing 0.1 or 0.4% human recombinant FGF-2 was topically applied to the defects and compared with no treatment. Eight weeks after application, the periodontal regeneration in those defects was analyzed. In all FGF-2-treated sites, significant periodontal regeneration was dose-dependently observed in greater amounts than in the carrier-treated or non-treated sites. No instances of epithelial down-growth, ankylosis, or root resorption were observed in the FGF-2-treated sites. These results indicate that a topical application of FGF-2 can enhance considerable periodontal regeneration in surgically created furcation class II defects of non-human primates.

Alveolar Bone Loss↗
PubMed2001

Diagnostic significance of surfactant proteins A and D in sera from patients with radiation pneumonitis.

Radiation pneumonitis (RP) is the most common complication of radiotherapy for thoracic tumours. The aim of this study was to evaluate the significance of pulmonary surfactant proteins (SP)-A and SP-D as new serum markers for RP. Twenty-five patients with lung tumour, who had received radiotherapy, were studied. At the completion of radiotherapy, the presence of RP was judged by chest plain radiography and chest high resolution computed tomography (HRCT). RP findings detected on chest plain radiography were seen in only three of 12 patients in whom RP was detected by HRCT. Nevertheless, both SP-A and SP-D concentrations in sera from the patients with RP were significantly higher than those from the 13 patients without RP (p = 0.0065, p = 0.0011, respectively). As with SP-A, ratios of SP-D at the completion, compared to at the initiation (1 week post/pre ratio), were also significantly higher in patients with RP than in patients without RP. When a post/pre ratio > 1.6 was considered positive, the SP-A and SP-D assays showed an 83% and 85% specificity, respectively. In conclusion, serum assays of surfactant proteins A and D may be of diagnostic value for detection of radiation pneumonitis, even when the radiographic change is faint.

Adult↗
PubMed2001

Malignant granular cell tumour of the cervical sympathetic nerve trunk.

Granular cell tumour is a rare neoplasm that can occur in various sites. This report describes a 48-year-old female with a malignant granular cell tumour originating from the cervical sympathetic nerve trunk, who presented with hyperaesthesia of the left shoulder. The tumour had a maximum diameter of 86 mm and involved adjacent tissues directly. Histopathologically, it showed considerable variation in the size and shape of nuclei, with occasional mitosis and an abundant granular cytoplasm. Immunohistochemically, the tumour reacted positively for S-100 protein and neuron-specific enolase, indicating its neural origin. Following total local excision of the lesion, the patient has been well without recurrence.

Autonomic Nervous System Diseases↗
PubMed2001

Production of catechol from benzoate by the wild strain Ralstonia species Ba-0323 and characterization of its catechol 1,2-dioxygenase.

Ralstonia sp. Ba-0323, a wild strain isolated from soil, produced catechol from benzoate and accumulated it outside the cells. The bacterium produced a maximal amount of catechol (1.6 mg/ml) from 3 mg/ml of sodium benzoate in a 20-h growing culture. The conversion rate of benzoate to catechol was 70% on a molar basis. The catechol production by the resting cells increased in the presence of glycerol, and the maximal amount of catechol produced from 3 mg/ml of sodium benzoate reached 1.9 mg/ml at the conversion rate of 83% after 8 h of incubation. Catechol 1,2-dioxygenase, which catalyzed the ring cleavage of catechol, was purified to homogeneity from a cell extract of Ralstonia sp. Ba-0323 growing on benzoate and characterized. The specific activity of the purified enzyme was much lower than those of the dioxygenases from other microorganisms reported. The Km for catechol of the purified enzyme was much higher than those of other dioxygenases. In addition, the NH2-terminal amino acid sequence of the enzyme was less similar to the other catechol 1,2-dioxygenases than they are to each other.

Amino Acid Sequence↗
PubMed2001

Regulation by two CatR proteins that differ in binding affinity to catB promoters expressing two cat gene clusters.

We isolated the two LysR-type regulatory proteins CatR1 and CatR2, which regulate the expression of cat1 and cat2 gene clusters, respectively, required for catechol degradation in the bacterium Frateuria sp. ANA-18. In a gel mobility shift assay using CatR1 and the DNA fragment containing the catB1 promoter region, the formation of two complexes, complex 1-1 (C1-1) and complex 1-2 (C1-2), was observed in the presence of cis,cis-muconate. On the other hand, CatR2 and the DNA fragment containing the catB2 promoter region formed only complex 2-2 (C2-2) at a lower concentration of cis,cis-muconate than that at which C1-1 and C1-2 were formed. As the concentration of cis,cis-muconate decreased, the production of the muconate cycloisomerase isozyme MC II encoded by catB2 decreased as well as that of MC I encoded by catB1. However, the amount of MC II synthesized was larger than that of MC I at low concentrations. On the basis of these results, we concluded that the catB2 promoter was activated at low concentrations of cis,cis-muconate.

Bacterial Proteins↗
PubMed2001

Purkinje cells in the adult cat cerebellar cortex possess a perineuronal net of proteoglycans.

The Purkinje cells in the adult cat cerebellar cortex were found to possess perineuronal proteoglycans which could be stained with our fine cationic iron colloid and Fujita's highly concentrated aldehyde fuchsin, and digested by chondroitinase ABC/keratanase/ heparitinase and hyaluronidase. The Purkinje cells are surrounded by some collagenous elements which are stained with Gömöri's ammoniacal silver and digested by collagenase. The Purkinje cells also express nerve cell surface glycoproteins which are labeled with lectin Vicia villosa agglutinin and digested by a double treatment with collagenase and endo-alpha-N-acetylgalactosaminidase. Sole digestion by endo-alpha-N-acetylgalactosaminidase never erased the lectin labeling of the nerve cell surface glycoproteins. These findings suggest that the collagenous elements mediate the linkage of the perineuronal proteoglycans to the nerve cell surface glycoproteins. It is presumed that in mice and rats, the perineuronal nets of proteoglycans and nerve cell surface glycoproteins of the Purkinje cells are so thin or coarse that they can not be sufficiently visualized under the light microscope.

Animals↗
PubMed2001

A sensitive method for detecting Porphyromonas gingivalis by polymerase chain reaction and its possible clinical application.

BACKGROUND: It is useful for the clinical diagnosis of periodontitis to monitor the colonization of periodontopathic bacteria in periodontal pockets. In this study, we attempted to establish and possibly identify the clinical application of a sensitive method to detect Porphyromonas gingivalis (P.g.), one of the putative periodontopathic bacteria related to chronic periodontitis. METHODS: Genomic DNA extracted from cultured P.g. 381 and clinically isolated subgingival plaque samples were used as a template of polymerase chain reaction (PCR). We designed primers to amplify the genomic DNA coding 40 kDa outer membrane protein (OMP), one of the unique proteins to all strains of P.g. The efficiency and specificity of amplification were evaluated by agarose gel electrophoresis and subsequent Southern hybridization with a digoxygenin-labeled oligonucleotide probe. RESULTS: Fewer than 100 P.g. bacterial cells in the specimen were reproducibly detected by PCR-hybridization assay. This PCR-hybridization assay was at least 100 times more sensitive than the conventional indirect immunofluorescence assay (IIF). Furthermore, the imaging analysis showed that there is a linear correlation between the strength of the signal and the cell number of P.g. from which the template DNA was extracted semiquantitatively. It is noteworthy that the PCR assay could also be applied to detect P.g. from clinical plaque samples and that it was approximately 100 times more sensitive than a conventional IIF assay. CONCLUSION: The PCR assay established in this study can be a powerful tool to detect P.g. in periodontal pockets and monitor the colonization and/or recolonization of P.g. at the very early phase.

Bacterial Outer Membrane Proteins↗
PubMed2001

Clinical characteristics of B-cell lymphoma-associated hemophagocytic syndrome (B-LAHS): comparison of CD5+ with CD5- B-LAHS.

OBJECTIVE: B-cell lymphoma-associated hemophagocytic syndrome (B-LAHS) has been increasingly reported in Asia and is regarded as a variant of intravascular lymphomatosis (IVL). Recently CD5 was reported to be expressed in some cases of diffuse large cell lymphoma and IVL. We therefore examined the expression of CD5 on lymphoma cells in B-LAHS and compared the clinical and laboratory data between CD5+ and CD5- B-LAHS. METHODS: The expression of CD5 on lymphoma cells was examined using flow cytometry and immunohistochemical analysis of paraffin sections. The clinical records were reviewed to characterize clinical features. PATIENTS: Twelve patients with B-LAHS; ten men and two women, age ranging from 41 to 82 years (median, 63.5 years) were included in this study. RESULTS: B-LAHS is characterized by fever and hepatosplenomegaly without lymphadenopathy at the initial presentation. Histological examination showed hemophagocytosis and infiltration of lymphoma cells in the bone marrow, and in some cases intravascular proliferation of lymphoid cells characteristic of IVL. All patients showed increased levels of lactate dehydrogenase, C-reactive protein, ferritin and soluble interleukin-2 receptor. In eight of the twelve patients, lymphoma cells were positive for CD5. But no differences were observed in the clinical or laboratory findings between CD5+ B-LAHS and CD5- B-LAHS. CONCLUSION: No clinical differences were observed between CD5+ B-LAHS and CD5- B-LAHS. Further studies are required to elucidate the differences in pathogenesis between these two subgroups of B-LAHS.

Adult↗
PubMed2001

Cupular response and functional anatomy to pressure application in the pigeon.

In this study, horizontal semicircular canal of the pigeon was inserted with a microfill, 164 micrometers in outer diameter, to gauge the pressure change in the canal and to clarify two things. One is whether the endolymph passes through the crista ampullaris or not. The other is as to the space where the endolymph passes through in the crista ampullaris if the endolymph goes through the crista. The pressure applications were performed by using Evans blue solution and as a result three pressure groups, i.e., 1) high, 2) moderate and 3) low were discriminated. Our results suggest that the endolymphatic fluid passes through the subcupular space or the top of the crista ampullaris when the pressure is enough small like the head rotates in daily life. Accordingly we present a new hypothesis as to the functional anatomy of the cupula in contrast to current hypotheses e.g. drum membrane or revolving door.

Animals↗
PubMed2001

The impact of a 4 cm. cutoff point for stratification of T1N0M0 renal cell carcinoma after radical nephrectomy.

PURPOSE: The 1997 TNM classification defines T1 tumors as those smaller than 7 cm. Recently, a cutoff point of 4 cm. has been proposed to create a subclass of T1 tumors. We evaluated the validity of this cutoff point by assessing the pathological findings and prognoses of patients with T1N0M0 renal cell carcinoma following radical nephrectomy. MATERIALS AND METHODS: We reviewed the hospital charts of 333 patients with T1N0M0 tumors, followed as long as 282 months (median 63) after radical nephrectomy. The validity of tumor size cutoff point for predicting survival outcome was tested in relation to other prognostic factors, including patient age, tumor position, nuclear grade, tumor histopathology and degree of microscopic venous invasion. RESULTS: During followup 32 patients (9.6%) had tumor recurrence and 21 (6.3%) died of renal cell carcinoma. A 5 cm. cutoff point maximized the differences in cancer specific survival rates and a 4 cm. cutoff point maximized the differences in disease-free survival rates. Tumor size was directly related to microscopic venous invasion and nuclear grade, which are significant prognostic factors, and a 4 cm. cutoff point enhanced these relationships. CONCLUSIONS: Tumor size is an important prognostic factor for patients with T1N0M0 renal cell carcinoma. A cutoff point of 4 cm. is practical for dividing the T1N0M0 classification into T1a and T1b subclasses.

Adult↗
PubMed2001

[Analysis on the relationship between use of health check ups and medical care by elderly patients. A study on urban cities with high health check-up rates].

The aim of this study was to evaluate the influence of health examination provided by municipalities in Japan on the use of medical care by elderly patients aged 70 and above. The study was carried out to compare health indicators of cities with high and low usage rates for health check ups over 14 years. The indicators of health service utilization included mean bed days, mean inpatient fees, and mean outpatient fees in 1983, 1988, 1993 and 1996. The data for these indicators were collected from all 12 Japanese cities, where more than 50% of the target population had annual health check-ups in 1983, 1986, 1989 and 1992, as the subjects of high rate cities. All other 134 cities belonging to the same prefectures with the 12 cities were included as the control low rate cities. The correlation coefficients for all the 146 cities showed that usage rates of health check ups had an inverse relationship with mean bed days and mean inpatient fees for the elderly population in all the fiscal years 1983, 1988, 1993 and 1996. In nine out of the ten prefectures, mean bed days and mean inpatient fees were lower in the high rate cities as compared to the low rate cities for all the fiscal years compared. The bed days in the 12 high rate cities were 72% of those in 134 low rate cities in 1983, and the percentages were 66%, 72%, and 78% in 1988, 1993 and 1996 respectively. No remarkable differences in mean outpatient fees were observed between the high and low rate cities. The differences in use of medical care by inpatients suggest that providing preventive health services would decrease the demand by the elderly and result in a more efficient use of health resources.

Aged↗
PubMed2001

[A clinicopathological evaluation of prognostic factors of urothelial tumors of the renal pelvis and ureter].

We investigated the clinicopathological features of 109 patients with urothelial tumors of the renal pelvis and ureter who underwent surgery at four institute from April, 1975 to September, 1997. The patients consisted of 71 males and 38 females, and the mean age was 66.8 years, ranging from 41 to 92 years. Mean followup period was 43 months. The prognostic significance of the pathological factors; pT, Grade, INF, tumor size, pL, pV and pN were evaluated. All these factors affected the survival rates significantly in univariate analysis. Multivariate analysis showed the most influential prognostic factors to be pT and pN.

Adult↗
PubMed2001

[Issues in cancer pain management].

In this article, three aspects of recent issues in cancer pain management such as pain assessment, drowsiness with morphine, and problem in home care setting. First, the assessment of 'impact of pain' is to assess whether the treatment is sufficient for the patient. On the other hand, assessment of 'pain intensity' is the effectiveness of the treatment. Therefore, to reduce the gap in pain evaluations between cancer patients and medical stuffs by assessing the 'impact of pain' in addition to 'intensity of pain'. Second, increasing dosage of morphine often reduces pain, at the same time patients become drowsy. This is often observed soon after starting morphine administration, especially, patients who have renal dysfunction. An active morphine metabolite morphine-6-Glucuronide is excreted into urine, so that, renal dysfunction causes drowsiness. Third, at home care setting, patients tend to be into poor pain control condition. It is necessary that paying attention on patient's knowledge of medications and having support program for pain management for home care setting.

Analgesics, Opioid↗
PubMed2001

The extracellular matrix in the mature brain: its reactions to endo-alpha-N-acetylgalactosaminidase and collagenase.

The hippocampal subiculum in the adult rat brain contains many neurons with nerve cell surface glycoproteins which are linked by collagenous ligands to perineuronal proteoglycans. The nerve cell surface glycoproteins or their terminal N-acetylgalactosamines are digested by endo-alpha-N-acetylgalactosaminidase. The terminal N-acetylgalactosamines linked by the collagenous ligands are not digested by endo-alpha-N-acetylgalactosasminidase. The collagenous ligands associated with the terminal N-acetylgalactosamines were digested by collagenase. The newly exposed terminal N-acetylgalactosamines by this collagenase incubation were digested by endo-alpha-N-acetyl-galactosaminidase. These findings on the rat agree with those obtained in our previous studies of the adult mouse brain samples. Furthermore they emphasize our hypothetical model that the perineuronal proteoglycans are related--via collagen ligands--with the nerve cell surface glycoproteins.

Animals↗