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Biomedical subjects

M Ogawa

Publications and source records attributed to M Ogawa.

At least 55 records · Page 3Linked to original sources

Induction of experimental ulcerative colitis by Fusobacterium varium isolated from colonic mucosa of patients with ulcerative colitis.

BACKGROUND: Bacteria are implicated in certain forms of model chronic colitis but the identity and role of bacteria in human ulcerative colitis (UC) are uncertain. AIMS: To isolate pathogenic bacteria from inflamed mucosa of patients with UC, to examine whether the bacteria have a toxin to Vero cells, and to determine whether the toxin induces UC-like lesions in animals. METHODS: Bacteria were isolated from UC patients and supernatants from cultures were filtered and tested for cytotoxicity to Vero cells. Bacterial cells producing the cytotoxic supernatants were examined by polymerase chain reaction for verotoxin genes. Culture supernatants of cytotoxic strains were examined by high performance liquid chromatography for organic acid concentrations. Mice were given enemas containing organic acid at the mean concentration in the supernatants of cytotoxic strains to ascertain whether colonic lesions appear in UC. RESULTS: Only supernatants from cultures of Fusobacterium varium killed Vero cells. Bacterial cells lacked verotoxin genes. Bacterial culture supernatants contained high concentrations of n-butyric acid and the mean concentration (32 mmol/l) was cytotoxic to Vero cells. Twenty four hours after mice were given enemas containing either butyric acid or F varium culture supernatants, colonic ulcers with crypt abscesses, inflammatory cell infiltration, and apoptotic changes were observed. CONCLUSIONS: Butyric acid in culture supernatants from cultures of F varium caused UC-like lesions in mice. This study indicates that F varium may be one of the elusive pathogenic factors in UC.

Adult↗

Surgical management of synchronous hepatocellular carcinoma and gastric cancer.

BACKGROUND/AIMS: The improvement of diagnostic modalities and operative techniques has resulted in prolonged survival for cancer patients, but has also led to the diagnosis of an increasing number of patients with synchronous hepatocellular carcinoma (HCC) and extrahepatic primary cancer. It is necessary to determine the optimal surgical strategies for synchronous HCC and gastric cancer. METHODS: In this retrospective study, clinicopathologic findings, diagnostic methods, treatment and outcome were reviewed in 13 patients who underwent curative surgery for synchronous HCC and gastric cancer. RESULTS: Twelve of the 13 patients were men older than 60 years. All patients had chronic hepatic disease, and hepatitis viral infection was detected in 9 patients. Examinations of the esophagus to search for esophageal varices before liver resection for HCC, and imaging studies to rule out liver metastasis before gastrectomy for gastric cancer can lead to the incidental finding of a synchronous carcinoma. The most frequent postoperative complication was massive ascites, which occurred in 4 patients who underwent lymph node dissection, 1 of whom died of perioperative hepatic failure. HCC recurred in 7 patients, 4 of whom died of their disease; only 1 patient died of recurrence of gastric cancer. CONCLUSION: Careful follow-up for recurrence of HCC is necessary because the most common cause of death in patients with synchronous carcinoma is recurrence of HCC.

Aged↗

[Bilateral coronary ostial stenoses with aortic regurgitation in a patient with syphilitic aortitis].

Syphilitic aortitis is now rare in developed countries and is sometimes overlooked. A 61-year-old man with bilateral coronary ostial stenoses (#5:90%, #1:99%) and Sellers III/IV aortic regugitatioin (AR) induced by syphilitic aortitis presented with chest pain. Preoperative rapid plasma reagin titer and Treponema pallidum hemagglutination test were strongly positive, 256 fold and 191.25 C.O.I., respectively. Aortic valve replacement (AVR) and coronary artery bypass grafting (CABG) with bilateral internal thoracic arteries (ITA) was performed successfully. The angiographic features as follows: 1) coronary artery stenosis is generally limited to the ostia, 2) the grade of stenosis is almost always more than 90%, 3) AR is frequently associated with coronary ostial stenosis. CABG should be performed with ITA, not saphenous vein grafts, to avoid occlusion of the ostium of the saphenous vein graft by syphilitic aortitis. Retrograde cardioplegia should be performed if ostial stenosis is severe.

Aortic Valve↗

[Combined operation for left ventricular aneurysm and ventricular septal perforation with normal coronary arteries; report of a case].

We experienced a rare case of left ventricular aneurysm (LVA) and ventricular septal perforation (VSP) with normal coronary arteries, and had a good outcome by undergoing Dor procedure and direct closure of VSP individually. A 68-year-old female was printed out cardiac systolic murmur. Echocardiogram revealed LVA and VSP, but coronary arteriogram showed no significant stenosis. The apex was incised longitudinally, lateral and parallel to the interventricular groove. VSP was directly closed by 2 pieces of 3-0 polypropylene suture with felt strip. Purse-string stitch with 3-0 polyester suture around the transitional zone between fibrotic tissue and viable tissue was secured to firm fibrous tissue. The residual orifice was closed with an elliptical composite patch of equine pericardial patch and polyester velour with 3-0 polypropylene suture. We considered based on the operative finding that myocardial infarction (MI) led to LVA and VSP. Coronary thrombosis rather than coronary spasm may have provoked MI because spasm provocative test was negative.

Aged↗

Identification of genes related to invasion and metastasis in pancreatic cancer by cDNA representational difference analysis.

The difference in gene expression between the highly invasive and metastatic cell line PC-1.0 and the weakly invasive and metastatic cell line PC-1 both derived from a pancreatic ductal carcinoma induced by N-nitrosobis (2-oxopropyl) amine (BOP) in Syrian golden hamster was examined using the Representational Difference Analysis (RDA) method. Within 8 clones (cDNA fragments) successfully isolated after subtraction hybridization of PC-1 cDNA from PC-1.0, 5 clones were shown to be specific or highly expressed in PC-1.0 as confirmed by RT-PCR. Among these 5 clones, two known genes, MAP kinase kinase 2 (MKK2) and phosphatidylinositol 4-kinase (PI4K230) were detected by gene sequencing. The specific expressions of MKK2 and PI4K230 in the highly invasive and metastatic cell line PC-1.0 were confirmed by RT-PCR and Northern blotting. By adding the CM of PC1.0 which included the cancer cell dissociation factor (DF), PC-1 cells began to dissociate and migrate from the colonies, and in addition the expression of MKK2 was found to be induced. On the other hand, the expression of PI4K230 was not induced in PC-1 cells by adding the CM of PC-1.0. Interestingly, in PC-1.0 the expression of PI4K230 was completely abolished and apoptosis induced by the PI3K inhibitor wortmannin. These results suggest that both MKK2 and PI4K230 are factors of a signal transduction pathway that might play an important role related to invasion and metastasis through the induction of cell motility and/or the inhibition of apoptosis.

1-Phosphatidylinositol 4-Kinase↗

Identification of early-responsive genes associated with the hypersensitive response to tobacco mosaic virus and characterization of a WRKY-type transcription factor in tobacco plants.

In order to identify genes that are involved in the defense reaction against pathogen attack, we screened for examples that are regulated during the hypersensitive response (HR) to infection by tobacco mosaic virus (TMV) of tobacco ( Nicotiana tabacum cv. Xanthi nc) carrying the N gene, which confers resistance to TMV. Among seven genes initially identified by fluorescent differential display, one clone was further characterized because its transcripts accumulated rapidly and transiently after the onset of HR. Its full-length cDNA of 1346 bp encoded a polypeptide consisting of 258 amino acids. The deduced protein contained a single WRKY domain, a Cys(2)His(2) zinc-finger motif and a leucine-zipper motif, showing high similarity to WIZZ, a member of the family of WRKY transcription factors in tobacco. The gene was thus designated TIZZ. A GFP-TIZZ fusion protein was found to localize to the nucleus upon introduction into epidermal cells of onion. Bacterially expressed TIZZ was able to bind to the W-box (TTGAC) element that is recognized by other WRKY proteins, but transactivation assays showed it to be unable to activate reporter gene expression by itself. TIZZ transcripts were induced in TMV-infected nahG transgenic tobacco plants, in which salicylic acid fails to accumulate. Neither exogenously applied salicylic acid nor mechanical wounding induced TIZZ transcript accumulation. These results indicate the presence of salicylic acid-independent pathways for HR signal transduction, in which a novel type of WRKY protein(s) may play a critical role for the activation of defense.

Amino Acid Sequence↗

Isolation and characterization of Lactococcus piscium strains from vacuum-packaged refrigerated beef.

AIMS: To characterize gram-positive, catalase-negative, psychrotrophic, lactic acid-homofermentative, non-motile cocci isolated from vacuum-packaged refrigerated beef using phenotypic and genotypic methods. METHODS AND RESULTS: A total of 89 strains was isolated at 2 and 6 weeks as one of the predominant microflora of five samples of vacuum-packaged beef stored at 2 degrees C. The strains were compared with reference strains of some gram-positive, catalase-negative cocci using SDS-PAGE whole-cell protein pattern analysis, biochemical characterization and 16S rDNA sequencing. The biochemical and physiological characteristics of the isolates resembled those of Lactococcus piscium GTC 552(T). Numerical analysis of the SDS-PAGE whole-cell protein patterns resulted in close clustering of the strains with L. piscium GTC 552(T) (r > 0.68). Other Lactococcus and Leuconostoc species could be distinguished from the isolates using SDS-PAGE whole-cell protein patterns (r < 0.58) and biochemical characteristics. The 16S rDNA sequencing of four randomly selected strains showed that the strains differed from L. piscium GTC 552(T) by two to three bases in the highly variable region of the sequence. This is the first report on the isolation of L. piscium from vacuum-packaged beef. CONCLUSIONS: The gram-positive catalase-negative cocci isolated from vacuum-packaged refrigerated beef have been identified as L. piscium. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this work contribute to the knowledge of the microflora of vacuum-packaged refrigerated beef.

Animals↗

Epithelial cell proliferation and apoptosis in the developing murine palatal rugae.

Epithelial cell proliferation and apoptosis during morphogenesis of the murine palatal rugae (PR) were examined histochemically by using anti-bromodeoxyuridine (BrdU) and the terminal deoxynucleotidyl transferase-mediated UTP nick-end-labelling (TUNEL) technique. Formation of the PR rudiment was observed as an epithelial placode in fetuses at 12.5 days post-coitus (dpc). During the PR formation, BrdU-positive cells were detected mainly in the epithelium of the interplacode and interprotruding areas in fetuses administered BrdU maternally at 2 h before killing. TUNEL-positive cells were detected only at the epithelial placode area in 12.5-14.5 dpc. At 16.5-18.5 dpc, the BrdU-positive cells were decreased in number in the epithelial cells at the interprotruding area of the PR. Only a few TUNEL-positive cells were observed in the protruding area of the PR at 16.5 dpc. These results suggest that cell proliferation and apoptosis in the palatal epithelium are involved spatiotemporally in the murine PR morphogenesis.

Animals↗

Growth-associated gene expression profiles by microarray analysis of trophoblast of molar pregnancies and normal villi.

We used microarray analysis to investigate expression profiles of 589 known genes committed to cell growth control to characterize regulatory circuitry for cell proliferation in complete moles (CMs). CMs are characterized by hyperplastic trophoblast and have a high propensity to give rise to choriocarcinoma. Characteristic alterations in gene expression profiles were observed when compared with normal villi. Fifty-seven genes were significantly up-regulated in CMs and involved the Ras-Map kinase 3, Jak-STAT5, and Wnt signal pathways, implicating growth factor or cytokine-mediated signal pathways in the trophoblastic hyperplasia of CMs. Several genes associated with anti-apoptosis, cell structuring, and/or cell attachment were also up-regulated in CMs. In contrast, relatively fewer genes were down-regulated and these involved IGFBPs, versican, interleukin-1, tumor necrosis factor receptor, CD44, and RAD52. Genes identified in this study may elucidate regulation mechanisms of trophoblastic proliferation and mechanisms causing a pathological phenotype in CMs.

Apoptosis↗

[Surgical repair of left ventricular free wall rupture using layered fibrin glue sheet and fibrin glue; report of a case].

A 57-year-old man with acute myocardial infarction (#13:90%, #6-#8:75%) was admitted to our hospital after the administration of tissue plasminogen activator. Three hours' after emergent percutaneous transluminal coronary angioplasty, he developed left ventricular free wall rupture in the left circumflex artery area. After bleeding was completely controlled by aortic cross clamping, a three-layered of fibrin glue sheet (TachoComb) with fibrin glue was extensively applied to the ruptured site including the infarcted area. He was discharged on the 25th postoperative day and underwent coronary artery bypass grafting to the left anterior descending artery three weeks later. This experience suggests that the layered TachoComb and fibrin glue are effective for left ventricular free wall rupture.

Coronary Artery Bypass↗

Assessment of the developmental totipotency of neural cells in the cerebral cortex of mouse embryo by nuclear transfer.

When neural cells were collected from the entire cerebral cortex of developing mouse fetuses (15.5-17.5 days postcoitum) and their nuclei were transferred into enucleated oocytes, 5.5% of the reconstructed oocytes developed into normal offspring. This success rate was the highest among all previous mouse cloning experiments that used somatic cells. Forty-four percent of live embryos at 10.5 days postcoitum were morphologically normal when premature and early-postmitotic neural cells from the ventricular side of the cortex were used. In contrast, the majority (95%) of embryos were morphologically abnormal (including structural abnormalities in the neural tube) when postmitotic-differentiated neurons from the pial side of the cortex were used for cloning. Whereas 4.3% of embryos cloned with ventricular-side cells developed into healthy offspring, only 0.5% of those cloned with differentiated neurons in the pial side did so. These facts seem to suggest that the nuclei of neural cells in advanced stages of differentiation had lost their developmental totipotency. The underlying mechanism for this developmental limitation could be somatic DNA rearrangements in differentiating neural cells.

Animals↗

Up-regulation of the ectodermal-neural cortex 1 (ENC1) gene, a downstream target of the beta-catenin/T-cell factor complex, in colorectal carcinomas.

To clarify the molecular mechanisms of human carcinogenesis associated with abnormal Wnt/wingless signaling, we searched for genes the expression of which was significantly altered by introduction of wild-type AXIN1 into LoVo colon cancer cells. By means of a cDNA microarray, we compared expression profiles of LoVo cells infected with either adenoviruses expressing wild-type AXIN1 (Ad-Axin) or those expressing a control gene (Ad-LacZ). Among the genes showing altered expression, the ectodermal-neural cortex 1 (ENC1) gene was down-regulated in response to Ad-Axin. The promoter activity of ENC1 was elevated approximately 3-fold by transfection of an activated form of beta-catenin together with wild-type T-cell factor (Tcf)4 in HeLa cells. Semiquantitative reverse transcription-PCR experiments revealed that expression of ENC1 was increased in more than two-thirds of 24 primary colon cancer tissues that we examined compared with corresponding noncancerous mucosae. Introduction of exogenous ENC1 increased the growth rate of HCT116 colon cancer cells in serum-depleted medium. In other experiments, overexpression of ENC1 in HT-29 colon cancer cells suppressed the usual increase of two differentiation markers, in response to treatment with sodium butyrate, a differentiation-inducible agent. These data suggest that ENC1 is regulated by the beta-catenin/Tcf pathway and that its altered expression may contribute to colorectal carcinogenesis by suppressing differentiation of colonic cells.

Axin Protein↗

Arg660Ser mutation in Thermus aquaticus DNA polymerase I suppresses T-->C transitions: implication of wobble base pair formation at the nucleotide incorporation step.

We examined the replication fidelity of an Arg660Ser (R660S) mutant of Thermus aquaticus DNA polymerase I (Taq pol I). In a forward mutation assay, R660S showed a marked reduction in T-->C transitions, one of the most frequent errors made by the wild-type enzyme. Steady-state kinetics showed that R660S discriminates against dGTP incorporation at a template T 13-fold better than the wild-type. R660S was also 3.2-fold less efficient than the wild-type at extending a T:dG mismatch. These results indicate that R660S has enhanced fidelity during incorporation and extension, which reduces its T-->C transition frequency. Interestingly, R660S also discriminated correct from incorrect nucleotides at the incorporation step of C:dATP, A:dATP, G:dATP and C:8-OH-dGTP mispairs 28-, 6.0-, 4.1- and 6.8-fold better, respectively, than the wild-type, although it may not always be as accurate as the wild-type at the extension step. A structural model suggests that Arg660 may participate in two interactions that influence fidelity; the guanidinium group of Arg660 might interact with the incoming guanine base at the major groove and it might compete for forming another interaction with the primer terminus. Substituting Arg with Ser may eliminate or alter these interactions and destabilize the closed complex with incorrect substrates. Our data also suggest that T:dGTP and C:dATP base pairs form 'wobble' structures at the incorporation step of Taq pol I.

Arginine↗

Bone marrow origin of hematopoietic progenitors and stem cells in murine muscle.

It has been reported that mononuclear cells harvested from murine skeletal muscle are capable of hematopoietic reconstitution of lethally irradiated mice. First, the nature of the hematopoietic progenitors in the muscle of C57BL/6-Ly-5.1 mice was examined by means of methylcellulose culture. The types and incidences of colonies grown from muscle mononuclear cells were different from those cultured from bone marrow (BM) or peripheral blood mononuclear cells. The next step was to examine the origin of the hematopoietic progenitors and stem cells in the muscle with the use of Ly-5.2 mice that had been made chimeric by transplantation of Ly-5.1 BM cells. The percentages of Ly-5.1 cells cultured from the muscle of the chimeric mice correlated with those cultured from BM, indicating BM origin of hematopoietic progenitors in the muscle. Long-term hematopoietic engrafting cells in the muscle of the chimeric mice were also derived from BM. However, mobilization of progenitors into circulation by granulocyte colony-stimulating factor did not change the population of hematopoietic progenitors in the muscle. It is proposed that hematopoietic progenitors and stem cells in the muscle tissue are of BM origin but their transition from BM to muscle may be a slow process.

Animals↗