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Biomedical subjects

T Sugai

Publications and source records attributed to T Sugai.

At least 37 records · Page 2Linked to original sources

Sebaceous gland metaplasia in intraductal papilloma of the breast.

We report here the first case of sebaceous gland metaplasia arising within an intraductal papilloma of the breast of a 70-year-old female. Several lobules and nests composed of clear cells closely resembling sebaceous glands of the skin were discovered within an intraductal papilloma of the breast. Squamous metaplasia was also noted in certain areas of the tumor. Immunohistochemically, the cells of the lobules and nests stained positively for monoclonal antibodies anti-cytokeratin 14 and epithelial membrane antigen. This study confirms a novel type of metaplasia of the breast.

Aged↗

Synthesis of methyl ursodeoxycholate via microbial reduction of methyl 7-ketolithocholate with Eubacterium aerofaciens JCM 7790 grown on two kinds of carbon and hydride sources, glucose and mannitol.

Eubacterium aerofaciens JCM 7790, which produces 7beta-hydroxysteroid dehydrogenase [7beta-HSDH], efficiently catalyzed the reduction of methyl 7-ketolithocholate [Me-7KLCA] to afford methyl ursodeoxycholate [Me-UDCA] in an anaerobic interface bioreactor. For the high accumulation of Me-UDCA, the best supplement of carbon and hydride source to the ABCM medium (glucose content, 0.3 wt.%) was screened because glucose strongly repressed 7beta-HSDH production. The supplementation of cellobiose, mannitol, sodium gluconate, and sodium pyruvate was effective to improve the production of Me-UDCA, although many mono- and disaccharides, such as fructose, sucrose, and lactose, strongly repressed 7beta-HSDH activity. A phosphate buffer was used as the aqueous phase in the carrier to avoid the lowering of medium pH in the carrier caused by the accumulation of organic acids via fermentation by E. aerofaciens. Thus, the productivity of Me-UDCA was elevated by the combination of glucose and mannitol as carbon and hydride sources and by using a phosphate buffer.

Journal Article↗

Preparation of methyl ursodeoxycholate via microbial reduction of methyl 7-ketolithocholate in an anaerobic interface bioreactor.

An interface bioreactor, which is a device for the microbial transformation of water-insoluble substrates, was applied to an anaerobic bioconversion for the first time. Methyl 7-ketolithocholate [Me-7KLCA] was reduced with the human intestinal bacterium Eubacterium aerofaciens JCM 7790 in a convenient anaerobic interface bioreactor using a nutrient agar plate placed in a GasPak pouch. The resulting methyl ursodeoxycholate [Me-UDCA] is a precursor of ursodeoxycholic acid, which is used as a cholesterol gallstone-dissolving agent. The reaction conditions were optimized, and ABCM medium and dihexyl ether were selected as the best carrier and reaction solvent, respectively. The toxicity of the bile acid esters toward the human intestinal bacterium was effectively alleviated in the interface bioreactor, in which the maximal concentrations of Me-7KLCA and Me-UDCA in the dihexyl ether layer respectively reached to 12.0 and 6.1 g/l.

Journal Article↗

Mass production of pure gibberellin A1 by Phaeosphaeria sp. L487 and the fungal preparation of [U-13C]gibberellin A1.

The mass production of pure gibberellin A1 (GA1) by shake-culturing Phaeosphaeria sp. L487 was investigated. Its GA1 production was markedly influenced by natural nitrogen sources and NH4NO3. When the fungus was cultured in an 8% glucose-1.5% oatmeal-0.1% NH4NO3-0.5% KH2PO4-0.1% MgSO4 x 7H2O medium for 3 weeks, the amount of GA1 in the culture filtrate was up to ca. 200 microg/ml: the addition of safflower oil to the culture medium two weeks after inoculation prolonged the GA1-production period to produce 300 microg/ml. Further preparation of [U-13C]GA, as a tool for the analysis of a complex of GA1 and its binding protein was attempted by using the fungus. The fungal culture in a [U-13C]glucose-oatmeal medium gave 6 mg of crystalline 13C-enriched GA1. Its 13C-enrichment of ca. 75% and 1J(CC) values were determined by NMR spectrometry.

Ascomycota↗

Genetic alterations in DNA diploid, aneuploid and multiploid colorectal carcinomas identified by the crypt isolation technique.

Loss of heterozygosity (LOH) and microsatellite instability (MSI) commonly occur in colorectal carcinomas. However, the role of these genetic alterations in determining DNA ploidy status of tumors (diploid, aneuploid and multiploid) remains unclear. In the present study, we attempted to clarify the relationship between genetic alterations and DNA ploidy status. Crypt isolation coupled with DNA cytometric sorting and polymerase chain reaction assay (17 microsatellite markers) were used to study allelic losses and MSI in 59 colorectal carcinomas (diploid, 15; aneuploid, 10 and multiploid, 34). Of the 15 diploid carcinomas, 6 exhibited MSI in which allelic losses were rarely found. The other 9 diploid tumors mostly exhibited allelic losses, but none displayed MSI status. Whereas allelic losses frequently occurred in the aneuploid carcinomas and the aneuploid populations of multiploid carcinomas, they were rarely detected in the diploid populations of multiploid carcinomas. MSI status was not observed in aneuploid carcinomas nor in either population of multiploid carcinomas. Although multiploid carcinomas genetically resemble aneuploid carcinomas in the expression of the severe LOH phenotype, the genetic alterations seen in the diploid populations of multiploid carcinomas may differ from those of diploid carcinomas. Furthermore, all diploid, aneuploid and both the diploid and aneuploid fractions of the multiploid tumors that were non-MSI exhibited a high rate of LOH, suggesting that LOH is independent of the tumor's ploidy status.

Adenocarcinoma↗

DNA mapping of gastric cancers using flow cytometric analysis.

Although numerous studies of gastric cancers on DNA ploidy have been reported, differences in the degree of aneuploidy (DNA index, DI) during progression have not been identified. We attempted to chart the differences in DIs during progression to clarify the role of aneuploidy in gastric cancers. We classified the gastric cancers examined into intestinal (n = 88) and diffuse (n = 48) types, and then analyzed 136 gastric cancers (intramucosal cancer, 42; submucosal cancer, 39; advanced cancer, 55) by flow cytometry using multiple sampling. In addition, we examined the DNA ploidy pattern of mucosal and submucosal lesions using the same submucosal cancers to study the tumor progression in individual cancers. Intratumoral DNA differences in DNA ploidy were observed in both types of gastric cancers. In intestinal-type cancers, multiple subclones indicated by a different DI occurred during the early stage of gastric cancers, whereas in diffuse-type cancers, multiple subclones were found primarily in advanced cancers. Although the DI varied widely in early intestinal-type cancers between 1.0 and 2.0, in early diffuse-type cancers, the DI tended to be less than 1.2. However, in advanced stage gastric cancers, the DI distribution was similar for both histological types. In intestinal-type cancers, high DI (>1.3) aneuploidy was frequently found in mucosal lesions. In contrast, only low DI (<1.2) aneuploid clones were observed in mucosal lesions of diffuse-type cancers. The present results suggest that high DI aneuploid tumor clones in intramucosal cancers acquire invasive ability when they progress to submucosal cancers, whereas DNA aneuploidy itself plays an important role in submucosal invasion of diffuse-type cancers.

Adult↗

Access to Wieland-Miescher ketone in an enantiomerically pure form by a kinetic resolution with yeast-mediated reduction.

Both enantiomers of Wieland-Miescher ketone [3,4,8, 8a-tetrahydro-8a-methyl-1,6(2H,7H)-naphthalenedione], in a highly enantiomerically enriched form, became readily available by a newly developed kinetic resolution with yeast-mediated reduction. From a screening of yeast strains, Torulaspora delbrueckii IFO 10921 was selected. The collected cells of this strain, obtained by an incubation in a glucose medium, smoothly reduced only the isolated carbonyl group of the (S)-enantiomer, while the (R)-enantiomer remained intact. Starting from both enantiomers ( approximately 70% ee) prepared by an established proline-mediated asymmetric Robinson annulation, the reduction with T. delbrueckii gave the (R)-enantiomer (98% ee) and the corresponding alcohol (4aS,5S)-4,4a, 5,6,7,8-hexahydro-5-hydroxy-4a-methyl-2(3H)-naphthalenone (94% ee, 94% de) in preparative scale in nearly quantitative yields. An approach for the asymmetric synthesis of the Wieland-Miescher ketone was also successful. 2-Methyl-2-(3-oxobutyl)-1,3-cyclohexanedione, the prochiral precursor, was reduced with this strain to give a cyclic acetal form of (2S, 3S)-3-hydroxy-2-methyl-2-(3-oxobutyl)cyclohexanone, in a stereomerically pure form.

Ketones↗

Aberrations of the K-ras, p53, and APC genes in extrahepatic bile duct cancer.

BACKGROUND AND OBJECTIVES: The genetic alterations involved in extrahepatic bile duct (EHBD) cancer are poorly understood. Our aim was to identify aberrations of the K-ras, p53, and APC genes in EHBD cancer. METHODS: We investigated aberrations of these genes in 52 EHBD cancers using polymerase chain reaction (PCR) single-strand conformation polymorphism analysis, followed by direct sequence determination and a PCR restriction fragment length polymorphism assay. RESULTS: The K-ras, p53, and APC genes were mutated in 9.6%, 32.7%, and 0% of EHBD cancers, respectively. Loss of heterozygosity at the p53 and APC gene loci was identified in 15.6% and 38.5% of EHBD cancers, respectively. CONCLUSIONS: Our results suggest that an unknown suppressor gene on 5q other than the APC gene may be responsible for EHBD cancer.

Aged↗

Comparison of Sepsis-related Organ Failure Assessment (SOFA) score and CIS (cellular injury score) for scoring of severity for patients with multiple organ dysfunction syndrome (MODS).

OBJECTIVE: To evaluate the usefulness of cellular injury score (CIS) and Sepsis-related Organ Failure Assessment (SOFA) score for determination of the severity of multiple organ dysfunction syndrome (MODS). DESIGN: A prospective observational study. SETTING: A medical and surgical intensive care unit (ICU) of a teaching hospital. PATIENTS: Forty-seven consecutive MODS patients. MEASUREMENTS AND RESULTS: SOFA score and CIS were measured every day for 12 months for 47 MODS patients. Comparison was made of the SOFA score and CIS for usefulness in the scoring of severity of MODS in 26 survivors and 21 non-survivors. In addition, receiver operating characteristics (ROC) analysis was used to determine the usefulness of these two indexes as predictors of prognosis. No significant differences were found on admission between the survivors and non-survivors, but significant differences between the two subgroups (p < 0.001) were found in maximum value within 1 week after admission and maximum value during the course of treatment for both indexes. Analysis of changes after admission indicated that significant differences between survivors and non-survivors began to appear on day 3 of admission for both indexes; at that time SOFA score began to deteriorate in the non-survivors while CIS began to improve in the survivors. ROC analysis demonstrated that the area under the ROC curve was 0.769 for SOFA scores and 0.760 for CIS. CONCLUSIONS: Both SOFA score and CIS sequentially reflected the severity of MODS. Furthermore, they were comparable in diagnostic value as predictors of prognosis. These findings may indicate the possibility that MODS is a summation of effects of cellular injury. In addition, sequential evaluation of both SOFA score and CIS would provide a more accurate prediction of prognosis than conventional methods.

Aged↗

Brunner's gland adenoma with a focus of p53-positive atypical glands.

A submucosal tumor was resected endoscopically from the duodenal bulb in a 43-year-old man complaining of epigastric discomfort. The tumor, measuring 22 x 20 x 19mm, consisted mainly of Brunner's glands with no atypia. However, close histologic examination disclosed a focus of glands with cellular and structural atypia. The atypical glands showed staining by periodic acid-Schiff, alcian blue, and high iron-diamine methods. Mucin histochemistry was examined, and the atypical glands resembled the excretory ducts rather than the acinar cells of the tumor. Immunohistochemically, positivity for MIB-1 was high (38.0%), and p53-positive cells were detected sporadically in the atypical glands. These results indicated that the atypical glands probably represented a neoplastic lesion. Brunner's gland adenomas associated with foci of true neoplasm are very rare; only two cases, including one patient with microcarcinoid tumors, have been reported.

Adenoma↗

Microsatellite instability in gallbladder carcinoma: two independent genetic pathways of gallbladder carcinogenesis.

Although the genetic basis for gallbladder carcinogenesis has not been clarified, considerable evidence has shown that genetic alterations play an important role in the development and progression of human cancers. In this study, we analyzed 30 gallbladder carcinomas to investigate the role of genetic alterations in their tumorigenesis, and to study correlations with their clinicopathological features. Tissue samples were obtained from 30 patients with gallbladder carcinoma (11 men and 19 women; mean age, 62 years; age range, 38-80 years). Genomic DNAs were extracted from fresh tumor tissue. We examined loss of heterozygosity (LOH) in the p53, APC, DCC, RB, and NM23-H1 gene regions by polymerase chain reaction (PCR)-LOH assay using an automated fluorescent DNA sequencer employing four microsatellite markers (p53, APC, DCC, NM23-H1). Five additional microsatellite markers were used for the determination of microsatellite instability (MSI). LOH was found at p53 in 9 of 15 informative cases (60%), at DCC in 10 of 22 (45%), at APC in 5 of 15 (33%), at RB in 1 of 8 (13%), and at NM23-H1 in 1 of 15 (7%). MSI was observed in 5 of 30 cases (17%) in at least one chromosomal loci of these nine microsatellite markers. None of the patients with MSI-positive tumors showed lymph node metastasis, and there was an inverse correlation between MSI and the presence of LOH in gallbladder carcinoma. These results suggest that there are two independent genetic pathways in gallbladder carcinogenesis; that is, an MSI pathway and an LOH pathway.

Adult↗

Microsatellite instability and mutation of mitochondrial and nuclear DNA in gastric carcinoma.

BACKGROUND & AIMS: Microsatellite instability (MSI) in mitochondrial DNA (mtDNA) is observed in some colorectal carcinomas. We attempted to determine if mitochondrial MSI (mtMSI) and mutations occur in gastric carcinomas and if the mtMSI phenotype underlies specific clinicopathologic profiles. METHODS: Sixty-two gastric carcinomas (34 intestinal and 28 diffuse types) were investigated. Coding mutations in 8 different mitochondrial genes, mtMSI in a noncoding (C)n tract, and p53 gene mutations were examined by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis. MSI in nuclear DNA (nMSI) and loss of the p53 gene were examined using microsatellite markers. RESULTS: Ten of 62 (16%) carcinomas showed the mtMSI phenotype. Mitochondrial gene mutation was detected in 5 carcinomas, 4 of which also showed the mtMSI phenotype. There was a positive correlation between mtMSI and nMSI status. In intestinal carcinomas, mtMSI, nMSI, and p53 gene alterations were frequently detected from early to advanced stages. In diffuse carcinomas, both kinds of MSI were found in only advanced (subserosal or serosal invasion) carcinomas. Six of 7 carcinomas with the nMSI phenotype and all 5 carcinomas with mitochondrial coding mutations had a considerable intestinal-type tumor cell component. CONCLUSIONS: Mitochondrial gene mutations, which are associated with the mtMSI phenotype, may play a specific role in the tumorigenesis of intestinal-type gastric carcinomas.

Cell Nucleus↗

Novel subdivisions of the rat accessory olfactory bulb revealed by the combined method with lectin histochemistry, electrophysiological and optical recordings.

Wistaria floribunda agglutinin and peanut agglutinin were found to bind histochemically to the anterior and posterior regions, respectively, of the vomeronasal nerve and glomerular layers in the rat accessory olfactory bulb. Furthermore, Ricinus communis agglutinin showed strong binding to the anterior region of the vomeronasal nerve and glomerular layers, whereas it bound weakly and/or moderately to the rostral two-thirds of the posterior glomerular layer but not at all to the caudal one-third. This suggests that the posterior region is further divided into two subregions. An electrophysiological mapping study in sagittal slice preparations demonstrated that stimulation given within the anterior vomeronasal nerve layer elicited field potentials within the anterior region of the external plexiform layer, whereas shocks to the rostral two-thirds and the caudal one-third of the posterior vomeronasal nerve layer provoked field responses within the rostral two-thirds and within the caudal one-third of the posterior external plexiform layer, respectively, indicating that the posterior external plexiform layer is also divided into two subregions. Real-time optical imaging showed similar results as above, except that neural activity also spread into mitral cell layers. Furthermore, the most anterior and posterior ends of the neural activity evoked in the rostral two-thirds of the posterior region immediately adjoined the posterior border of that evoked in the anterior region and the anterior border of that evoked in the caudal one-third of the posterior region, respectively. Moreover, the granule cell layer was also found to have similar boundaries. Thus, optical imaging studies demonstrated individual precise boundaries of these subdivisions, which were positioned right beneath those defined by Ricinus communis agglutinin histochemistry. The presence of functional segregation in each layer leads us to conclude that there are at least three different input-output pathways in the rat vomeronasal system.

Animals↗

Use of crypt isolation to determine loss of heterozygosity of multiple tumor suppressor genes in colorectal carcinoma.

Analysis of loss of heterozygosity (LOH) is very important in the study of tumor suppressor genes. However, accurate LOH analysis of tumor suppressor genes is difficult because of dilution by contaminating non-tumor DNA. Thus, enrichment of tumor DNA is required to accurately determine LOH of the tumor. We developed a new application of the fluorescent polymerase chain reaction by coupling it with crypt isolation to accurately assess the incidence of LOH of tumor suppressor genes in 45 colorectal carcinomas. LOH was observed at p53 in 26 of 37 tumors (70.3%), at APC in 13 of 35 (37.1%), at DCC in 16 of 25 (64.0%), at NF-2 in 5 of 23 (21.7%), and at nm23 H-1 in 7 of 30 (23.3%). We could clearly determine LOH of these genes because the crypt isolation technique was used. Although the incidence of LOH at each of these loci, as determined by using this technique, was similar to that obtained in previous studies using conventional methods, this method provides a simpler, more accurate way to assess LOH. In addition, the morphology of the samples can be analyzed before genetic analysis.

Adenocarcinoma, Mucinous↗

Esophageal squamous cell carcinoma characterized by extensive chondroid differentiation.

A rare case of carcinoma characterized by extensive chondroid elements at a site of primary esophageal and metastatic lesion is reported. The patient was a 67-year-old man complaining of dysphagia due to an ulcerative lesion at the lower middle esophagus. He underwent irradiation treatment prior to surgery. Histologically, the tumor consisted of both carcinomatous and chondroid elements and had invaded deeply into the esophageal wall. The carcinomatous cells had gradually become chondroid cells embedded within an extensive extracellular matrix. In addition, the metastatic lesion showed findings similar to those of the primary lesion. Immunohistochemistry revealed that both carcinomatous and chondroid elements were immunostained with cytokeratin and epithelial membrane antigen, suggesting an epithelial nature to the chondroid cells. Conversely, only chondroid cells were positively stained for S-100 protein. Furthermore, bone morphogenetic proteins (BMP) were positive for chondroid cells and their surrounding carcinomatous cells. Given the apparent transition between carcinomatous and chondroid cells based on microscopy and immunohistochemical findings in the present case, we concluded that the chondroid cells were derived from carcinomatous cells. In addition, our findings suggest that BMP produced by carcinomatous cells lead to chondroid differentiation of the carcinoma cells.

Aged↗