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N Kamiya

Publications and source records attributed to N Kamiya.

At least 37 records · Page 2Linked to original sources
PubMed2000

Structural basis for the higher Ca(2+)-activation of the regulated actin-activated myosin ATPase observed with Dictyostelium/Tetrahymena actin chimeras.

Replacement of residues 228-230 or 228-232 of subdomain 4 in Dictyostelium actin with the corresponding Tetrahymena sequence (QTA to KAY replacement: half chimera-1; QTAAS to KAYKE replacement: full chimera) leads to a higher Ca(2+)-activation of the regulated acto-myosin subfragment-1 ATPase activity. The ratio of ATPase activation in the presence of tropomyosin-troponin and Ca(2+) to that without tropomyosin-troponin becomes about four times as large as the ratio for the wild-type actin. To understand the structural basis of this higher Ca(2+)-activation, we have determined the crystal structures of the 1:1 complex of Dictyostelium mutant actins (half chimera-1 and full chimera) with gelsolin segment-1 to 2.0 A and 2.4 A resolution, respectively, together with the structure of wild-type actin as a control. Although there were local changes on the surface of the subdomain 4 and the phenolic side-chain of Tyr230 displaced the side-chain of Leu236 from a non-polar pocket to a more solvent-accessible position, the structures of the actin chimeras showed that the mutations in the 228-232 region did not introduce large changes in the overall actin structure. This suggests that residues near position 230 formed part of the tropomyosin binding site on actin in actively contracting muscle. The higher Ca(2+)-activation observed with A230Y-containing mutants can be understood in terms of a three-state model for thin filament regulation in which, in the presence of both Ca(2+) and myosin heads, the local changes of actin generated by the mutation (especially its phenolic side-chain) facilitate the transition of thin filaments from a "closed" state to an "open" state. Between 394 and 469 water molecules were identified in the different structures and it was found that actin recognizes hydrated forms of the adenine base and the Ca ion in the nucleotide binding site.

Actins↗
PubMed2000

Appraisal of surgical resection of gallbladder cancer with special reference to lymph node dissection.

BACKGROUND: Radical lymph node dissection in surgery for advanced gallbladder cancer is controversial. The purpose of this study is to evaluate the role of lymph node dissection based on the clinico-pathologic results. PATIENTS: Seventy-three patients who underwent radical surgery including systematic dissection of the N1+N2 region lymph node plus some of the para-aortic nodes were reviewed. RESULTS: pT1 patients had no lymph node metastasis, but pT2 and pT3/pT4 patients had lymph node metastasis at a rate of 50.0% (13/26) and 83.3% (25/30), respectively. As infiltration of the hepatoduodenal ligament (Binf) became severe, the rate and extent of lymph node metastasis increased. There were four 5-year survivors with lymph node involvement. The 5-year survival rates are 77.0% in pN0 cases and 27.3% in pN1 cases (P<0.01). There was no difference in survival between pN1 and pN2 patients. However, significant differences in survival were observed between pN0/1 and pN2/3 patients when these patients were limited to Binf0/1. Examination of the recurrence pattern showed that most patients with pN0/1/2 had no regional lymph node recurrence, but there was para-aortic lymph node recurrence in patients with pN3 outside the dissected region. Significant prognostic factors influencing survival after surgery by multivariate analysis were pN2/3, pT, and residual tumor. CONCLUSION: Systematic lymph node dissection of N1, N2, and part of the para-aortic region improves survival in advanced gallbladder cancer patients, especially in those without either para-aortic lymph node metastases or Binf2/3.

Adult↗
PubMed2000

Gene expression and immunohistochemical localization of biglycan in association with mineralization in the matrix of epiphyseal cartilage.

This study has used in situ hybridization, Northern blot analysis, and immunohistochemistry at the light and electron microscope levels to localize mRNAs and core proteins of biglycan in developing tibial epiphyseal cartilage of 10-day old Wistar rats. The expression of mRNAs and core proteins of biglycan appeared prominent in hypertrophic and degenerative chondrocytes associated with the epiphyseal ossification centre and the growth plate cartilage, but was not seen in the rest of epiphyseal cartilage. Northern blot analysis confirmed biglycan mRNA expression in the epiphyseal cartilage. Ultrastructural immunogold cytochemistry of the growth plate revealed that prominent immunolabelling was confined to the Golgi apparatus and cisternae of rough-surfaced endoplasmic reticulum of the hypertrophic and the degenerating chondrocytes, the early mineralized cartilage matrices of the longitudinal septum of the lower hypertrophic and the calcifying zones, and fully mineralized cartilage mitrices, which were present in the metaphyseal bone trabeculae. Furthermore, Western blot analysis of biglycan in extracts of fresh epiphyseal cartilage revealed that an EDTA extract, after chondroitinase ABC digestion, contains core proteins of biglycan, indicating the presence of biglycan in mineralized cartilage matrices. These results indicate that the distribution of biglycan is associated with cartilage matrix mineralization.

Animals↗
PubMed2000

[The risk of colorectal adenomatous polyp in relation to serum total cholesterol levels in Japanese men classified by age group].

In order to investigate the relationship between the risk of colorectal cancer and serum total cholesterol (TC) levels, we studied the relationship between the risk of colorectal adenomatous polyp and high TC levels in Japanese men aged from 40 to 59 at a manufacturing company. We first diagnosed 283 subjects as having adenomatous polyp (AD) or normal (C), identified by means of an immunological fecal occult blood detection test and total colonofiberscopic examination (TCF), and then selected 94 subjects after excluding the patients with diseases possibly influencing TC. We classified the subjects into two age groups (40-49 and 50-59) and conducted a statistical analysis of AD and C groups by means of a two tailed t-test. The mean values for TC (DTC), which had been obtained within 3 months before TCF, were significantly different (p < 0.001 95% CI 15.79-48.49) for those in their 40s, but were not different for those in their 50s. In order to check the stability of the data, we analyzed the mean values for DTC and BTC, which had been obtained within 15 months. The results showed the same trend for those in their 40s (p = 0.001 95% CI 10.76-40.87). The mean DTC and BTC values were again not significantly different for those in their 50s. We further divided the subjects into the lower (-181 mg/dl) and higher (209 mg/dl-) DTC values and conducted further statistical analysis with Fisher's exact test. The results were significant in the 40-49 age group (p = 0.004), in the odds ratio (13.75 95% CI 2.32-81.49), but showed no significant difference in the 50-59 age group. The mean LDL-cholesterol values within 3 months before TCF were significantly different in the 40-49 age group (p < 0.001 95% CI 15.22-47.70) and were not different in the 50-59 age group. There was no statistical difference in the mean values for HDL-cholesterol, triglyceride and body mass index in either age group. From these results, it can be concluded that the risk of colorectal adenomatous polyp was associated with a higher serum total cholesterol level in the 40s, and seemed to indicate a close relationship with the risk of colorectal cancer.

Adenomatous Polyps↗
PubMed2000

[Studies of gastric acid secretion and gastric mucosal change after Helicobacter pylori eradication--the short and long-term observation].

We studied the gastric acid secretion and the histological findings of gastric mucosa in 48 patients with peptic ulcer before and after successful Helicobacter pylori (H. pylori) eradication. The pH of fasting gastric juice significantly decreased from 2.46 to 1.65 four weeks after eradication (p < 0.001). The factors such as disease, atrophic border, inflammatory cell involvement and histological atrophy were analyzed by logistic regression. Only the atrophic border was a significant and independent factor to convert the acidity after eradication, and the pH of gastric juice prominently decreased in the patients with widespread atrophic mucosal area. In the 24-hour intragastric pH monitoring, the pH3 holding time overnight remarkably decreased from 54.1% to 22.3% after eradication (p < 0.005). In the long follow-up of 17 patients (the mean follow-up period was 15.2 months), the pH of gastric juice kept a lower titer of 1.66. The mean atrophic scores in the upper body reduced from 1.5 to 0.9 (p < 0.05). In addition to the functional recovery of parietal cells after eradication, it was considered that the histological improvement of atrophy in oxyntic mucosa was associated with the recovery of pH.

Adult↗
PubMed2000

[Multidisciplinary treatment of hilar cholangiocarcinoma].

With advances in surgical techniques, the number of curative resection for hilar cholangiocarcinoma has increased. However, the recurrence rate after curative resection is significantly high. There is no established adjuvant therapy for these patients, although some groups have administered intraarterial infusion of 5-fluorouracil combined with cisplatin. In our institution, postoperative radiation is administered to control remnant tumors at the ductal and surgical margins. Extracorporeal radiation is directed at the surgical margins and intraluminal radiation at the ductal margins. This has improved the cumulative survival rate for cholangiocarcinoma patients who underwent both curative and noncurative resection. The 3-year survival rate of those who underwent curative resection with and without postoperative radiation is 100% and 28.5%, respectively. The longest survival duration among our patients who received intraluminal radiation for positive ductal margins were 55 months. Because the biological behaviour and radiation sensitivity of the tumors differ among individually, these should be determined through molecular biologic studies in the future. Such tailored therapy will require multidisciplinary treatment.

Bile Duct Neoplasms↗
PubMed1999

Enantioselective recognition mechanism of secondary alcohol by surfactant-coated lipases in nonaqueous media.

The enantioselective recognition mechanism of secondary alcohol by lipases originated from Candida rugosa and Pseudomonas cepacia was elucidated on the basis of the kinetic study of the esterification of alcohol with lauric acid in isooctane. To obtain inherent kinetic parameters, we utilized a surfactant-coated lipase whose conformation is considered to be an "open" form in a homogeneous organic solvent. Based on the experimental results, the enantioselectivity of lipases was found to be derived from the difference in the V(max) values between the two enantiomers. The same result was observed when lipases of different origin and substrates with different molecular structures were applied. &copy 1999 John Wiley & Sons, Inc.

Alcohols↗
PubMed1999

Surfactant-histidine-heme ternary complex as a simple artificial heme enzyme in organic media

A surfactant-heme complex which shows peroxidase activity in organic media has been prepared by a method utilizing water-in-oil (W/O) emulsions. Both the aqueous phase pH and the type of surfactant appeared to have prominent effect on the catalytic activity of the heme complex in benzene. The catalytic efficiency of the heme complex was enhanced more than ten times by adding histidine to the aqueous phase of W/O emulsions in the preparation process. The enhancement of peroxidase activity was observed only in a nonaqueous medium due to the increase of the effective concentration of histidine as an activator. In the present study, we propose a simple preparation method for an artificial heme enzyme which works in nonaqueous media. Copyright 1999 John Wiley & Sons, Inc.

Journal Article↗
PubMed1999

Tertiary and quaternary structures of photoreactive Fe-type nitrile hydratase from Rhodococcus sp. N-771: roles of hydration water molecules in stabilizing the structures and the structural origin of the substrate specificity of the enzyme.

The crystal structure analysis of the Fe-type nitrile hydratase from Rhodococcus sp. N-771 revealed the unique structure of the enzyme composed of the alpha- and beta-subunits and the unprecedented structure of the non-heme iron active center [Nagashima, S., et al. (1998) Nat. Struct. Biol. 5, 347-351]. A number of hydration water molecules were identified both in the interior and at the exterior of the enzyme. The study presented here investigated the roles of the hydration water molecules in stabilizing the tertiary and the quaternary structures of the enzyme, based on the crystal structure and the results from a laser light scattering experiment for the enzyme in solution. Seventy-six hydration water molecules between the two subunits significantly contribute to the alphabeta heterodimer formation by making up the surface shape, forming extensive networks of hydrogen bonds, and moderating the surface charge of the beta-subunit. In particular, 20 hydration water molecules form the extensive networks of hydrogen bonds stabilizing the unique structure of the active center. The amino acid residues hydrogen-bonded to those hydration water molecules are highly conserved among all known nitrile hydratases and even in the homologous enzyme, thiocyanate hydrolase, suggesting the structural conservation of the water molecules in the NHase family. The crystallographic asymmetric unit contained two heterodimers connected by 50 hydration water molecules. The heterotetramer formation in crystallization was clearly explained by the concentration-dependent aggregation state of NHase found in the light scattering measurement. The measurement proved that the dimer-tetramer equilibrium shifted toward the heterotetramer dominant state in the concentration range of 10(-2)-1.0 mg/mL. In the tetramer dominant state, 50 water molecules likely glue the two heterodimers together as observed in the crystal structure. Because NHase exhibits a high abundance in bacterial cells, the result suggests that the heterotetramer is physiologically relevant. In addition, it was revealed that the substrate specificity of this enzyme, recognizing small aliphatic substrates rather than aromatic ones, came from the narrowness of the entrance channel from the bulk solvent to the active center. This finding may give a clue for changing the substrate specificity of the enzyme. Under the crystallization condition described here, one 1,4-dioxane molecule plugged the channel. Through spectroscopic and crystallographic experiments, we found that the molecule prevented the dissociation of the endogenous NO molecule from the active center even when the crystal was exposed to light.

Amino Acid Sequence↗
PubMed1999

Effects of bone morphogenetic protein-2 and transforming growth factor-beta1 on gene expression of decorin and biglycan by cultured osteoblastic cells.

The influence of bone morphogenetic protein-2 (BMP-2) and transforming growth factor beta (TGF-beta) on the expression of small proteoglycans, decorin and biglycan was investigated in a clonal rat osteoblastic cell line, ROS-C26 (C26) cells, which is a potential osteoblast precursor cell line and capable of differentiating into mature osteoblasts after treatment with recombinant BMP-2 (rhBMP-2). Following the culture of C26 cells for 3, 6, and 9 days in the presence or absence of rhBMP-2, alkaline phosphatase activity increased in the rhBMP-2 treated cells in direct proportion to their differentiation into more mature osteoblastic cells, whereas decorin mRNA decreased in the cells, when compared to control cells without rhBMP-2 treatment. These results were evident 6 days after treatment. However, rhBMP-2 treatment had no effect on biglycan mRNA expression in the cells. Subsequently, after removal of rhBMP-2 from the culture media, the cells were further cultured for 24 h with graded concentrations of TGF-beta1 (0, 0.1, 1.0, 5.0, and 10 ng/ml). TGF-beta1 decreased decorin mRNA expression in the cells dose dependently, but did not affect their biglycan mRNA expression. Furthermore, either removal of rhBMP-2 from the culture media or addition of TGF-beta1 significantly decreased alkaline phosphatase activity of rhBMP-2-induced cells. These results indicate that osteoblastic differentiation is accompanied by increased alkaline phosphatase activity and decreased expression of decorin mRNA, but continuous expression of biglycan mRNA. Both rhBMP-2 and TGF-beta1 inhibit decorin mRNA expression in osteoblasts at varying stages of differentiation, but their effects on biglycan mRNA expression and alkaline phosphatase are different.

Alkaline Phosphatase↗
PubMed1999

Effects of transforming growth factor-beta1 on the gene expression of decorin, biglycan, and alkaline phosphatase in osteoblast precursor cells and more differentiated osteoblast cells.

In this study, the effects of incubating two clonal rat osteoblastic cell lines at different stages of differentiation, ROB-C26 (C26) and ROB-C20 (C20), with transforming growth factor-beta1 (TGF-beta1) on the gene expression of decorin, biglycan, and alkaline phosphatase were examined. C26 cells are a potential osteoblast precursor cell line that is also capable of differentiating into muscle cells and adipocytes and is differentiated into osteoblasts after treatment with bone morphogenetic protein-2. C20 cells are a more differentiated osteoblastic cell line. Our Northern blot studies demonstrated that after treatment with TGF-beta1 (0, 0.1, 1.0, 5.0, and 10 ng/ml), a dose- and time-dependent decrease in decorin mRNA expression was found in C26 cells. In contrast, the effect of decorin mRNA with TGF-beta1 was not determined in C20 cells, since decorin mRNA expression was extremely low in this cell line even in the absence or presence of TGF-beta1. Although TGF-beta1 treatment resulted in no appreciable effect on biglycan mRNA expression in both cell lines in a dose- and time-dependent manner, it decreased significantly the expression of alkaline phosphatase in both cell lines at the gene and protein level. Reverse transcriptase-polymerase chain reaction analysis revealed the gene expression of decorin, and TGF-beta type I and type II receptors in both cell lines. These results indicate that osteoblasts progenitor cells express both decorin and biglycan mRNAs. In contrast, more differentiated and mature osteoblastic cells express preferentially biglycan mRNA. TGF-beta1 exerts different effects on the expression of decorin and biglycan mRNAs, and is a potent inhibitor of the gene expression of alkaline phosphatase during osteoblast differentiation.

Alkaline Phosphatase↗
PubMed1999

Assessment of DNA content in formalin-fixed, paraffin-embedded tissue of lung cancer by laser scanning cytometer.

A new cytometric device, a laser scanning cytometer was developed to overcome the limitations of flow cytometry (FCM) and image analyses. The purpose of this study was to develop a method that allows laser scanning cytometry (LSC) to be used for measuring the cellular DNA content of paraffin-embedded tissues. Paraffin-embedded lung cancer tissue from 30 patients was analyzed by both FCM (p-FCM) and LSC (p-LSC). In addition, touch preparations from fresh frozen tissues were prepared to provide material for LSC (f-LSC). The limits of agreement for the DNA indices (DI) measured by p-LSC and p-FCM were -0.07 to 0.07, indicating that for a given case, these methods would be expected to differ by no more than 0.07. The limits of agreement for comparisons between the other materials and methods were wider and depended upon the size of the measurements. Agreement between f-LSC and p-FCM was good for small DI values, but poor for large values. Agreement between f-LSC and p-LSC was poor for small and large DI values, but good for moderately sized values. Discordancies in DNA ploidy status between different materials and methods may have been caused by either the heterogeneity within tumors, sampling errors or differences in the interpretation of histograms. This method allows a comparison of the results of DNA analysis with histologic findings from hematoxylineosin-stained sections and the prognosis of the patients.

Aneuploidy↗
PubMed1999

Endobronchial brachytherapy for recurrent thymoma showing endobronchial polypoid growth.

The authors report a case of recurrent thymoma displaying endobronchial polypoid growth. Initially, the patient had invasive thymoma with intracaval growth into the right atrium. He was treated with multimodality therapy consisting of chemotherapy, surgical resection, and radiotherapy (50.4 Gy). Both 3 years and 6 years after the initial treatment, the tumor recurred outside the reconstructed superior vena cava. The patient was treated with repeated radiotherapy (50.4 Gy and 40 Gy), and remission was achieved. Eight years after the first therapy, an endobronchial polypoid lesion was detected in the right upper lobe bronchus and was histologically found to be thymoma. Endobronchial high-dose rate brachytherapy (20 Gy at 3 mm/5 fractions) was carried out for palliation because the recurrent tumor occurred outside of the superior vena cava area, which had been reirradiated. After the treatment, the endobronchial tumor shrunk remarkably in size without adverse effects. No tumor regrowth has been noted after a follow-up of 10 months.

Aged↗
PubMed1999

Specific lipid-protein interactions in a novel honeycomb lattice structure of bacteriorhodopsin.

In the purple membrane of Halobacterium salinarium, bacteriorhodopsin trimers are arranged in a hexagonal lattice. When purple membrane sheets are incubated at high temperature with neutral detergent, membrane vesicularization takes place, yielding inside-out vesicles with a diameter of 50 nm. The vesicular structure becomes unstable at low temperature, where successive fusion of the vesicles yields a crystal which is composed of stacked planar membranes. X-ray crystallographic analysis reveals that the bacteriorhodopsin trimers are arranged in a honeycomb lattice in each membrane layer and that neighbouring membranes orient in opposite directions. The native structure of the trimeric unit is preserved in the honeycomb lattice, irrespective of alterations in the in-plane orientation of the trimer. One phospholipid tightly bound to a crevice between monomers in the trimeric unit is suggested to act as a glue in the formation of the trimer.

Bacteriorhodopsins↗
PubMed1999

Detection of brain metastasis in potentially operable non-small cell lung cancer: a comparison of CT and MRI.

STUDY OBJECTIVE: To compare the usefulness of MRI and CT in the detection of brain metastases during preoperative evaluation and postoperative follow-up. DESIGN: A prospective and sequential comparison. PATIENTS AND METHODS: Of 332 patients with potentially operable non-small cell lung cancer who were free of neurologic signs and symptoms, brain CT was performed preoperatively on 155 patients (CT group) and brain MRI on 177 patients (MRI group). Patient characteristics in both groups were comparable. In 279 patients with complete resection of the primary lung tumor, intensive follow-up with CT and MRI was performed in the respective groups. The preoperative detection of brain metastases, postoperative intracranial recurrence rates, and characteristics of detected brain tumors were compared between the two groups. The survival of patients with brain metastases was also compared. RESULTS: From the first evaluation to 12 months after surgery for primary lung cancer, brain metastases were observed in 11 patients (7.1%) from the CT group and 12 patients (6.8%) from the MRI group. MRI detected brain metastases preoperatively in 6 of the 12 patients (3.4% of the total MRI group), whereas CT detected brain metastases preoperatively in 1 of the 11 patients (0.6% of the total CT group). MRI showed a tendency toward a higher preoperative detection rate of brain metastases than CT (p = 0.069). Furthermore, the mean (+/- SD) maximal diameter of the brain metastases was significantly smaller in the MRI group (12.8+/-9.1 mm) than in the CT group (20.3+/-7.0 mm) (p = 0.041). However, the median survival time and 2-year survival rate after treatment of detected brain metastases, respectively, were 10 months and 27% in the CT group and 17 months and 28% in the MRI group. There was no significant difference between the groups in survival time. CONCLUSIONS: Preoperative evaluation and intensive follow-up with MRI could facilitate early detection of brain metastases in patients with potentially operable lung cancer. However, further studies on detection and treatment of the metastatic tumors are considered necessary.

Adenocarcinoma↗