Biomedical subjects

H Inui

Publications and source records attributed to H Inui.

At least 37 records · Page 2Linked to original sources

PubMed2000

Expression of human cytochromes P450 1A1 and P450 1A2 as fused enzymes with yeast NADPH-cytochrome P450 oxidoreductase in transgenic tobacco plants.

Among 11 isoforms of the human cytochrome P450 enzymes metabolizing xenobiotics, CYP 1A1 and CYP 1A2 were major P450 species in the metabolism of the herbicides chlortoluron and atrazine in a yeast expression system. CYP1A2 was more active in the metabolism of both herbicides than CYP1A1. The fused enzymes of CYP1A1 and CYP1A2 with yeast NADPH-cytochrome P450 oxidoreductase were functionally active in the microsomal fraction of the yeast Saccharomyces cerevisiae and showed increased specific activity towards 7-ethoxyresorufin as compared to CYP1A1 and CYP1A2 alone. Then, both fused enzymes were each expressed in the microsomes of tobacco (Nicotiana tabacum cv. Samsun NN) plants. The transgenic plants expressing the CYP1A2 fusion enzyme had higher resistance to the herbicide chlortoluron than the plants expressing the CYP1A1 fusion enzyme did. The transgenic plants expressing the CYP1A2 fused enzyme metabolized chlortoluron to a larger extent to its non-phytotoxic metabolites through N-demethylation and ring-methyl hydroxylation as compared to the plants expressing the CYP1A1 fused enzyme. Thus, the possibility of increasing the herbicide resistance in the transgenic plants by the selection of P450 species and the fusion with P450 reductase is discussed.

Atrazine↗

PubMed2000

Increases in gastric histidine decarboxylase activity and plasma gastrin level in streptozotocin-induced type 1 diabetic rats.

When type 1 diabetes mellitus was induced in rats by injecting streptozotocin, histidine decarboxylase expression was abnormally up-regulated in a transcriptional level, and 7 d after the injection, the enzyme activity was increased about 3-fold over the control (p < 0.05). When the diabetic rats were administered with insulin for 3 d, the increased histidine decarboxylase activity returned to a normal level in addition to normalization of the plasma glucose level. The plasma gastrin level in the fasting state was also significantly elevated in the diabetic rats, and the insulin treatment normalized the level. In the diabetic rats, fasting gastric acid output increased significantly and gastric pH was lowered. These results suggest that the gastric histidine decarboxylase activity and plasma gastrin level are increased in connection with the depletion of insulin in streptozotocin-induced diabetic rats, and gastric acid secretion is stimulated at a basal level, presumably due to increases in the concentrations of histamine and gastrin in oxyntic mucosa.

Animals↗

PubMed1999

Mutations of a novel human RAD54 homologue, RAD54B, in primary cancer.

Association of breast tumor susceptibility gene products BRCA1 and BRCA2 with the RAD51 recombination protein suggested that cancer could arise through defects in recombination. The identification of NBS1, responsible for Nijmegen breakage syndrome, from the MRE11/RAD50 recombination protein complex also supports this hypothesis. However, our mutation analysis revealed that known members of the RAD52 epistasis group are rarely mutated in human primary cancer. Here we describe the isolation of a novel member of the SNF2 superfamily, characterized with sequence motifs similar to those in DNA and RNA helicases. The gene, designated RAD54B, is significantly homologous to the RAD54 recombination gene. The expression of RAD54B was high in testis and spleen, which are active in meiotic and mitotic recombination. These findings suggest that RAD54B may play an active role in recombination processes in concert with other members of the RAD52 epistasis group. RAD54B maps to human chromosome 8q21.3-q22 in a region associated with cancer-related chromosomal abnormalities. Homozygous mutations at highly conserved positions of RAD54B were observed in human primary lymphoma and colon cancer. These findings suggest that some cancers arise through alterations of the RAD54B function.

Adenocarcinoma↗

PubMed1999

Mutations in the RAD54 recombination gene in primary cancers.

Association of a recombinational repair protein RAD51 with tumor suppressors BRCA1 and BRCA2 suggests that defects in homologous recombination are responsible for tumor formation. Also recent findings that a protein associated with the MRE11/RAD50 repair complex is mutated in Nijmegen breakage syndrome characterized by increased cancer incidence and ionizing radiation sensitivity strongly support this idea. However, the direct roles of BRCA proteins and the protein responsible for NBS in recombinational repair are not clear though they are associated with the recombinational repair complexes. Since RAD51 forms a complex with other members of the RAD52 epistasis group and with BRCA proteins, it is reasonable to ask if alterations of members of the RAD52 epistasis group lead to tumor development. Here we describe missense mutations at functional regions of RAD54 and the absence of the wild-type RAD54 expression resulting from aberrant splicing in primary cancers. Since RAD54 is a recombinational protein associated with RAD51, this is the first genetic evidence that cancer arises from a defect in repair processes involving homologous recombination.

Adenocarcinoma↗

PubMed1999

Disordered expression of the sucrase-isomaltase complex in the small intestine in Otsuka Long-Evans tokushima fatty rats, a model of non-insulin-dependent diabetes mellitus with insulin resistance.

To clarify the relationship between diabetes mellitus and carbohydrate digestion, the activities of sucrase and isomaltase, which form a complex enzyme (SI complex) on the brush border membranes, were compared in the progression of diabetes mellitus in Otsuka Long-Evans Tokushima fatty (OLETF) rats, a model of human non-insulin-dependent diabetes mellitus with insulin resistance, and Long-Evans Tokushima Otsuka (LETO) rats as non-diabetic controls. Until 40 weeks of age, OLETF rats were obese and had a high plasma glucose level, compared to age-matched LETO rats, but the sucrase and isomaltase activities showed no significant differences between the two strains. Oral glucose tolerance test revealed that during 40-48 weeks of age, NIDDM became very severe with advancing insulin resistance in OLETF rats. In OLETF rats, in contrast to LETO rats, at 48 weeks of age, abnormal increases in the sucrase and isomaltase activities occurred, along with a remarkable decrease in body weight and a further great increase in the plasma glucose level in the non-fasting state. Hyperinsulinemia occurred in 20-week-old OLETF rats; however, at 40 and 48 weeks of age, the plasma insulin level in the non-fasting state in OLETF rats was not significantly different from that in LETO rats. The level of mRNA encoding the SI complex increased abnormally in 48-week-old OLETF rats. These results suggest that the advance of insulin resistance leads to an increase in the expression of the SI complex on the transcriptional level.

Age Factors↗

PubMed1999

Comparison of p53 and bcl-2 expression in initial, synchronous, and metachronous colorectal adenomas.

To investigate the significance of p53 and bcl-2 expression in metachronous colorectal adenomas arising in the remaining colon after carcinoma resection, we analyzed p53 and bcl-2 expression immunohistochemically in initial adenomas (type I), synchronous adenomas with concurrent carcinoma (type II), and metachronous adenomas arising after resection of initial adenomas (type III) or carcinoma (type IV). The incidence of p53 immunoreactivity in type IV adenomas with mild dysplasia was significantly higher than that in type I, type II, or type III adenomas with mild dysplasia. bcl-2 immunoreactivity was more frequently detected in type IV adenomas with mild dysplasia than in type I or type III adenomas with mild dysplasia. Coexpression of p53 and bcl-2 was detected in 16% of the type IV adenomas, this being a significantly higher frequency than that seen in the type I, type II, or type III adenomas. These results suggest that the evaluation of p53 and bcl-2 in metachronous adenomas in the remaining colon after resection of carcinoma may be a useful biologic marker for assessing the risk of cancer development.

Adenoma↗

PubMed1999

Purification and characterization of ADP-ribosyl cyclase from Euglena gracilis.

ADP-ribosyl cyclase, which catalyzes the conversion from NAD+ to cyclic adenosine diphosphoribose (cADPR), is proposed to participate in cell cycle regulation in Euglena gracilis. This enzyme, which was found as a membrane-bound protein, was purified almost the homogeneity after solubilization with deoxycholate, and found to be a monomeric protein with a molecular mass of 40 kDa. Its Km value for NAD+ was estimated to be 0.4 mM, and cADPR, a product of the enzyme, inhibited the enzyme competitively with respect to NAD+ whereas another product, nicotinamide, showed noncompetitive (mixed-type) inhibition. In contrast to mammalian CD38 and BST-1, Euglena ADP-ribosyl cyclase lacked cADPR hydrolase activity.

ADP-ribosyl Cyclase↗

PubMed1999

Acceptance of skin allografts in pigs by portal venous injection of donor bone marrow cells.

OBJECTIVE: To confirm in pigs whether a new method for organ allografts, originally established in mice by the authors, might be applicable to humans. SUMMARY BACKGROUND DATA: The authors recently established a new method for organ allografts in mice that includes the injection of donor bone marrow cells (BMCs) using the portal vein (PV), followed by the administration of cyclosporin A (CsA) on days 2 and 5, and the intravenous injection of BMCs on day 5. In the present study, they modify this method (a single-day protocol) and apply it to pigs. METHODS: Allogeneic BMCs of donor pigs were injected using the PV (a superior mesenteric vein). The skin grafting was carried out on the day of the PV injection. The recipient pigs received donor grafts, autologous grafts, and third-party grafts at the same time. In addition, an open wound was made as the epithelized control. Full-thickness skin grafts were harvested from the dorsal wall of the donors. CsA (10 mg/kg) was injected intramuscularly into recipient pigs on days 2 and 5 after the PV injection. RESULTS: One hundred percent of skin grafts survived for >300 days when donor BMCs were injected using the PV (n = 6). However, the skin grafts of the three pigs that had received BMCs using the intravenous route were rejected within 3 to 4 weeks after transplantation. The third-party skin grafts showed necrotic changes on day 21 after transplantation. CONCLUSIONS: One hundred percent of skin allografts can be obtained, even in pigs, by injecting donor BMCs using the PV, carrying out skin allografts, and administering CsA on days 2 and 5. This single-day protocol would be of great advantage for human organ transplantation.

Animals↗

PubMed1999

[Vitamin B1].

Vitamin B1 (thiamin), taken-up into cells, is converted to thiamin diphosphate (TDP), and TDP acts as a cofactor for several enzymes involving in carbohydrate metabolism. CoA-dependent oxidative decarboxylation of pyruvate is catalyzed by pyruvate dehydrogenase multienzyme complex (PDC) with NAD+ as an electron acceptor in most organisms involving mammals and higher plants. PDC consists of three component enzymes, one of which is pyruvate dehydrogenase (lipoamide) which contains TDP as a prosthetic group. Similar multienzyme complex for 2-oxoglutarate or branched chain 2-oxoacids is also found in mammals. In anaerobic bacteria, archaebacteria and anaerobic protozoa, pyruvate:ferredoxin oxidoreductase (PFOR) functions for the oxidative decarboxylation of pyruvate with ferredoxin in place of NAD+. PFOR contains TDP as a cofactor; however its structure is quite different from PDC and 1-3[4Fe-4S] clusters are involved as redox centers. Pyruvate:NADP+ oxidoreductase (PNOR), which catalyzes the oxidative decarboxylation of pyruvate with NADP+ as an electron acceptor, occurs in mitochondria of Euglena gracilis, a protist containing chloroplasts. PNOR consists of two functional domains, one of which contains TDP and 3[4Fe-4S] clusters and resembles PFOR. Another domain involves FMN and FAD as redox centers and its structure is similar to NADPH-cytochrom P450 reductase.

Carboxy-Lyases↗

PubMed1998

The intron 5-inserted form of rat erythropoietin receptor is expressed as a membrane-bound form.

The cDNA encoding an intron 5-inserted form of the erythropoietin receptor (I5Epo-R) has been cloned from rat. DNA sequence analysis reveals that the insertion of intron 5, which consists of 79 bp, causes a shift in reading frame and results in termination in the region of exon 7. The deduced amino acid sequence is composed of 316 amino acid residues, which is a molecular weight of 34220. To study the function of rat I5Epo-R, we established a Chinese hamster ovary cell line expressing rat I5Epo-R. Western blot analysis and binding studies with 125I-recombinant human erythropoietin showed that the transfected cells expressed rat I5Epo-R with a molecular size of 36 kDa as a membrane-bound form, but not as a soluble form, and had a single class of binding sites with a Kd of 700 pM.

Alternative Splicing↗

PubMed1998

Suppressive effect of insulin on the synthesis of sucrase-isomaltase complex in small intestinal epithelial cells, and abnormal increase in the complex under diabetic conditions.

An abnormally high level of the sucrase-isomaltase (SI) complex in the small intestine of rats with streptozotocin-induced insulin-dependent diabetes mellitus (IDDM) was normalized in 11 h by the administration of insulin, in addition to normalization of the blood glucose level. Phlorizin, an inhibitor of renal glucose reabsorption, also caused normalization of the blood glucose level in the IDDM rats; however, the level of the SI complex was barely changed. When mucosa explants were cultured in a medium, the SI complex synthesized during the cultivation was accumulated as its precursor protein without maturation, owing to the absence of pancreatic proteases, and the amount of the precursor protein that accumulated in the explants was decreased by the addition of insulin into the medium. Further, the mRNA level of the SI complex in the explants incubated with insulin was obviously lower than that in the absence of insulin. These results indicate that insulin has a suppressive effect on the synthesis of the SI complex, presumably by decreasing the transcriptional level of the gene encoding the complex, in small-intestinal epithelial cells. Thus the synthesis of the SI complex might exceed normal levels in the epithelial cells as a direct result of the depletion of insulin under IDDM conditions.

Animals↗

PubMed1998

Managing bile duct injury during and after laparoscopic cholecystectomy.

Laparoscopic cholecystectomy is now the treatment of choice for gallstones, but there has been concern that bile leakage after a laparoscopic cholecystectomy is more frequent than after an open cholecystectomy. We have experienced 16 patients with bile duct injury after a laparoscopic cholecystectomy. Five patients had a circumferential injury to the major bile duct, and we employed a converted open technique for biliary reconstruction. The other 11 patients had partial injury to the major bile duct, and we performed laparoscopic restoration; all 11 of these patients received endoscopic retrograde cholangiography (ERC) on the day after the operation and stenting for biliary decompression and drainage. No complications were identified and the duration of hospitalization in these patients was significantly shorter than in those who had the converted procedure. If intraoperative cholangiography is performed routinely, the presence and form of bile duct injury can be clearly identified, and the decision to restore the site of injury or to convert to the open technique for biliary reconstruction can be made immediately.

Adult↗

PubMed1998

Induction chemoradiotherapy for advanced stage III non-small cell lung cancer: long-term follow-up in 42 patients.

This multi-institutional phase II study was designed to assess the feasibility, efficacy, toxicity, and long-term survival of induction chemoradiotherapy followed by surgery in previously untreated patients with advanced stage III non-small cell lung cancer. Chemotherapy regimen included cisplatin 20 mg/m2 on days 1-5 and 29-33, and VP-16 40 mg/m2 on days 1-5 and 29-33. Radiotherapy (50 Gy in 25 fractions) began on day 1. Clinically downstaged patients underwent thoracotomy 3-5 weeks after the completion of radiotherapy. Forty-two eligible patients (ten stage IIIA and 32 IIIB) were followed for a median period of 64 months. The response rate was 81%, and 20 patients had a clinically good response. Twenty-one patients underwent thoracotomy. Nineteen patients had complete resections and there were seven pathologic complete responses. There were four treatment related deaths (all stage IIIBs). There were significant survival differences between stage IIIA versus IIIB patients (P = 0.028; median survivals, 24.9 vs. 11.1 months; 5-year survival rates, 20% vs. 8.3%), and patients that achieved pathologic complete response (CR) versus those that did not (P = 0.045; median survivals 30.1 vs. 11.1 months; 5-year survival rates, 28.6% vs. 8.3%). Although the induction chemoradiotherapy employed in this study was not appropriate for stage IIIB patients, it proved feasible in stage IIIA patients in whom it resulted in good 5-year survival rates. It also provided good survival rates in patients achieving pathologic CR.

Adult↗

PubMed1998

Hemodynamics of a simulation of the vertebrobasilar system using silicone tube.

We investigated the hemodynamics of the vertebrobasilar system (VBS) using a simulation model tube made of silicone which had different diameters between the bilateral vertebral artery (VA) ducts (stenotic and normal VA ducts). Using the ultrasonic Doppler method. the laterality index (L.I.) of the flow velocity in the VA duct was 32.88%. The flow volume of the anterior inferior cerebellar artery duct and the posterior inferior cerebellar artery duct also had significant laterality. By investigating the effects of the rotatory pulse rate and fluid viscosity, a maximal decrease to below 40-50% of the flow volume of the branches, and to below 71.93% in the mean flow velocity of the basilar artery (BA) duct (BA duct) were observed. and the maximum L.I. increased to 43.15%. When the unilateral stenotic VA duct was occluded by clamping (clamping occlusion), the flow volume profiles of the branches were the same as without clamping occlusion. The L.I. showed no significant changes and the maximal decrease in the mean flow velocity of the BA duct was 68.61%. Using the laser Doppler method, the flow velocity distribution of the BA duct was shifted towards the side of the stenotic VA duct. These results suggest that the rheological dynamics in the main vessels can certainly reflect the posterior circulatory dynamics, and that modulating factors also aggravate the hemodynamics of the VBS with this disorder.

Arteries↗

PubMed1998

Preoperative assessment for laparoscopic cholecystectomy: feasibility of using spiral computed tomography.

OBJECTIVE: The authors investigated the preoperative feasibility of using spiral computed tomography (SCT) after intravenous infusion cholangiography (IVC-SCT) for laparoscopic cholecystectomy. SUMMARY BACKGROUND DATA: In laparoscopic cholecystectomy, the aberrant or unusual anatomy of the bile duct and severe inflammation or adhesions around the gallbladder sometimes require a conversion to open surgery. METHODS: Laparoscopic cholecystectomies (LC's) were attempted on 440 patients, and preoperative IVC-SCT also was attempted in all of these patients. Using this spiral scanning technique, the bile ducts, cystic duct, and gallbladder were assessed for contour abnormalities, relative position, and filling defects. Forty-seven patients were diagnosed with having stones in their common bile duct or common hepatic duct. RESULTS: Three-hundred eighty-seven patients out of the 440 patients (88.0%) who were subjected to IVC-SCT had the length and course of their cystic duct successfully determined. Anomalous unions of the cystic duct were seen in 59 (15.2%) of 387 patients with respect to the operative findings, and 48 of 440 patients (10.9%) had severe adhesions to Calot's triangle and the surrounding tissues. In these 48 patients, 45 patients (94%) had a nonvisualized cystic duct on IVC-SCT. The preoperative assessment of the feasibility (dense adhesions obscuring Calot's triangle) of using IVC-SCT demonstrated that the sensitivity, specificity, and accuracy were 93%, 98%, and 94%, respectively. Five patients had to be converted to open surgery, and the overall morbidity rates for patients undergoing laparoscopic cholecystectomy was 0.9% (4 of 440). CONCLUSIONS: The most important factor in assessing the feasibility of using laparoscopic cholecystectomy is not the nonvisualized gallbladder, but the nonvisualized cystic duct on IVC-SCT. IVC-SCT may be of benefit to those patients scheduled to undergo laparoscopic cholecystectomy.

Adolescent↗

PubMed1998

Infrequent mutations in the PTEN/MMAC1 gene among primary breast cancers.

Recently PTEN/MMAC1, a candidate tumor suppressor gene, was isolated from chromosome 10q23-24 and somatic mutations of this gene were detected in several malignancies including brain, prostate, and breast tumors. To investigate further the potential role of this gene in mammary carcinogenesis, we examined 69 primary breast cancers for mutations in PTEN/MMAC1 by means of polymerase chain reaction single-strand conformation polymorphism and sequencing analysis. We detected only one somatic missense mutation, a change from T to C at codon 59 (TCA to CCA) resulting in substitution of Pro for Ser in the predicted protein. This site is located outside of phosphatase or phosphate-acceptor motifs, but this codon encodes a residue that is conserved in homologous proteins, tensin and auxilin and is likely to be crucial for normal function of PTEN/MMAC1. Among the 69 tumors examined, three low-frequency polymorphisms were found as well, one in the non-coding region of exon 1 and one each in introns 2 and 7. Our results suggested that mutation of the PTEN/MMAC1 gene is not a major factor in the development of most primary breast cancers.

Adult↗

PubMed1998

Suppression by water extracts of Sophora plants of sucrose-induced hyperglycemia in rats and inhibition of intestinal disaccharidases in vitro.

Partially purified hot-water extracts of the roots of plants of the Sophora family suppressed the increase in blood glucose concentration of rats in the oral sugar tolerance test. The extracts also inhibited rat intestinal sucrase and maltase. The most potent sample was about 15 times more active than catechin, a positive control, in these experiments.

Animals↗

PubMed1998

Alpha 2-adrenoceptor-mediated antisecretory effect of hypoxia in conscious rats.

Gastric acid secretion is suppressed, resulting in a significant rise in gastric pH, when conscious rats are exposed to hypoxia (Yamaji et al., 1996). When adrenal medullectomized rats were exposed to moderate (10.5% O2) hypoxia for 3 h, gastric acid secretion was restored to nearly the level in normoxia by the adrenal medullectomy. In severe (7.6% O2) hypoxia, the operation also caused an increase in the level of gastric acid output, although the extent was lower than that under 10.5% O2 hypoxic conditions. Gastric pH was normalized by the operation even with 7.6% O2 hypoxia. Similar results were obtained when reserpine, which causes an adrenergic discharge, was administered. When an alpha 2-adrenoceptor blocking agent, yohimbine, was administered, the inhibitory effect of 10.5% and 7.6% O2 hypoxia on gastric acid secretion was almost completely removed. However, neither prazosin (an alpha 1-adrenoceptor blocker) nor propranolol (a beta-adrenoceptor blocker) showed any significant effects on gastric acid output in hypoxia. These results indicate that acute hypoxia stimulated the adrenergic response from the adrenal medulla, and that gastric acid secretion was consequently suppressed through alpha 2-adrenoceptor.

Adrenal Medulla↗