Biomedical subjects

T Hirota

Publications and source records attributed to T Hirota.

At least 163 records · Page 9Linked to original sources

PubMed1991

Fluorimetric and high-performance liquid chromatographic determination of D-lactate in biological samples.

D-Lactate in biological samples was converted into a strongly fluorescent substance in a one-vial reaction. It was first converted into the pyruvate hydrazone in the presence of D-lactate dehydrogenase, an NADH-reoxidation system using diaphorase, D,L-6,8-thioctamide and hydrazine. This hydrazone was then converted into 2-hydroxy-6,7-dimethoxy-3-methylquinoxaline by 1,2-diamino-4,5-dimethoxybenzene in 1 M hydrochloric acid, and the quinoxaline was extracted and measured fluorimetrically at 432 nm (excitation at 365 nm). The calibration curve for D-lactate was linear up to at least 100 nmol/ml of the assay mixture, with a determination limit of 2 nmol/ml. The quinoxaline was also analysed by high-performance liquid chromatography with fluorimetric detection. The calibration curve for D-lactate was linear from 500 fmol to 75 nmol in the reaction mixture. This method was 4000 times more sensitive than the fluorimetric method, and could determine D-lactate in blood plasma volumes of less than 1 microliter.

Animals↗

PubMed1991

Lymph node metastasis in thoracic esophageal carcinoma.

Seventy-nine patients with thoracic esophageal carcinoma underwent transthoracic esophagectomy with neck, mediastinal, and abdominal lymphadenectomy. The operative mortality rate was 3.8%. Fifty-seven patients (72.2%) had metastasis in the lymph nodes. Though three patients with carcinoma classified as pTis had no positive nodes, nine (50.0%) of the patients with a pT1 carcinoma had positive nodes. The 5-year survival rate for 57 patients with positive nodes was 33.6%. Twenty-nine patients (36.7%) had positive nodes in the neck; 47 (59.5% ), in the mediastinum; and 33 (41.8%), in the abdomen. Their 5-year survival rates were 30.0%, 24.4%, and 38.4%, respectively. The differences between these rates were not statistically significant. These results indicate that the neck lymph nodes should be regarded as part of the regional lymph nodes and that esophagectomy with wide lymph node dissection improves the long-term survival of patients with thoracic esophageal carcinoma.

Abdomen↗

PubMed1991

Effect of Konjac mannan on intestinal microbial metabolism in mice bearing human flora and in conventional F344 rats.

The effects of dietary Konjac mannan (KM), a frequent ingredient of traditional Japanese foods, on intestinal microbial metabolism and microflora composition were investigated using two laboratory animal models, namely, conventional F344 rats and C3H/He male mice bearing human flora. Dietary KM led to a significant reduction in faecal beta-glucuronidase, nitroreductase and azoreductase activities, and in the production of phenol and indole in the faeces of conventional F344 rats. In the C3H/He male mice bearing human flora, faecal beta-glucuronidase and nitroreductase activities were significantly reduced by KM ingestion, as were the amounts of the putrefactive products, p-cresol and indole, in the faeces. Slight differences in intestinal microflora composition between control and KM diet groups were noted. The results indicate that, in C3H/He male mice bearing human flora, dietary KM may modify microbial metabolism without causing significant alterations in intestinal microflora composition.

Animals↗

PubMed1991

A possible error in the interpretation of gastric carcinoma.

Isolated clusters of pyloric cells having homogeneous, eosinophilic (i.e., "glassy") cytoplasm were present in 6 (18.2%) of 33 consecutive gastrectomy specimens. Clusters with "glassy" cells displayed either a glandular arrangement or appeared as cohesive clusters. Cohesive clusters could be misinterpreted as signet-ring cell adenocarcinoma. However, remnants of the apical mucous secretion from the "ordinary" pyloric cells, the nuclei usually near the free border of the cells, as well as the negative reaction in the "glassy" cytoplasm for neutral and acid mucins, helped to differentiate them from signet-ring adenocarcinoma cells.

Adenocarcinoma↗

PubMed1991

Point mutation of c-Ki-ras oncogene in gastric adenoma and adenocarcinoma with tubular differentiation.

The presence of point mutation at codons 12, 13 and 61 of the c-Ki-ras oncogene was investigated in 7 cases of gastric adenoma and 35 cases of gastric adenocarcinoma using DNA samples from formalin-fixed and paraffin-embedded tissues. Oligonucleotides encompassing the three codons were amplified by using the polymerase chain reaction (PCR), and then examined for point mutation by the selective oligonucleotide hybridization technique. Point mutation was detected in three of the 7 adenomas (43%) and three of the 35 carcinomas (9%). All the gastric adenomas showed the histology of tubular adenoma, being very similar to that of colonic adenoma. The 35 cases of gastric adenocarcinoma were classified into 17 cases of differentiated type and 18 cases of undifferentiated type including signet-ring cell carcinoma. The point mutation of c-Ki-ras oncogene was detected only in the differentiated type (3/17, 18%), and there was no case with point mutation in the undifferentiated type. These results suggest that the genetic mechanism of carcinogenesis differs between the differentiated type and the undifferentiated type of gastric adenocarcinoma, and also that c-Ki-ras activation is possibly involved in a relatively early step of the "adenoma-carcinoma sequence," which leads to the development of a portion of differentiated adenocarcinomas in the stomach.

Adenocarcinoma↗

PubMed1991

Alterations in copy number of c-erbB-2 and c-myc proto-oncogenes in advanced stage of human breast cancer.

In our previous study, amplification of c-erb B-2 and c-myc proto-oncogenes in DNA of human breast cancer occurred in 16% and 4% of cases, respectively, and increased copy number of these genes is suggested to be associated with aggressive primary tumors. We examined change in the copy number of c-erb B-2 and c-myc proto-oncogenes between primary and multiple metastatic tumors in 10 patients with breast cancer, who underwent breast surgery and were later autopsied, by using DNAs isolated from formalin-fixed paraffin-embedded tissues and the dot blot-hybridization method. In primary tumors, amplification of c-erb B-2 and c-myc was detected in three and two cases, whereas at the stage with systemic metastasis, it was detected in four and three cases, respectively. In all four cases with amplified c-erb B-2 gene and in one of the three with amplified c-myc gene, the copy number was clearly increased in the metastatic tumors in comparison with the primary. Microscopically, more than five mitotic figures per high power field were detected in metastatic tumors of five cases including three with amplified c-erb B-2, but in only two primary. These results suggested that the aggressive nature of breast cancer is frequently enhanced in accordance with cancer metastasis.

Breast Neoplasms↗

PubMed1991

Safety follow-up in a cohort of Biken acellular DPT vaccine recipients in Japan.

In October 1984 in Sweden, a phase II trial of Biken acellular Pertussis vaccine was started and in 1986, a phase III trial of the same vaccine was begun. During the phase III trial, there were three cases of deaths out of 1,385 of study children at two, four and ten weeks after the second dose of the vaccine, due to severe invasive bacterial infections such as H. influenzae, Pneumococcus, or Meningococcus infection. A number of arguments arose about the results of the Phase III trial. No one can either prove or disprove the association between invasive bacterial infection and administration of acellular pertussis vaccine. The purpose of this paper is to discuss the side effects of Biken acellular DPT vaccine. The pediatricians inquired about the physical status of the children who received Biken acellular DPT vaccine. During the observation period, three out of 940 infants suffered from infectious diseases. One suffered from measles, the other from varicella and the last from mumps. Our retrospective study did not reveal any severe invasive bacterial infection cases cases such as the ones experienced in Sweden.

Bacterial Infections↗

PubMed1991

Esophageal and pulmonary metastases from ovarian carcinoma: a case report of long-term survival following metastatic resections.

A case of ovarian carcinoma which has been followed up for more than 28 years after the initial ovarian resection is reported. A 50-year-old woman came forward with a metastatic esophageal tumor 16 years after a salpingo-oophorectomy for ovarian carcinoma. After irradiation with a dose of 3090 cGy, an esophagectomy and a partial resection of the directly invaded pericardium was successfully performed. Ten years later, progressive dyspnea due to a total collapse of the right lung developed. The right main stem bronchus was almost occluded by an endobronchial tumor, contiguous with the recurrent mass at the previous resectional margin. A right pneumonectomy and a combined resection of the pericardium and left atrium were performed. The patient has been followed up for further two years, and no signs of recurrence are evident. In spite of distant thoracic involvement and its local regrowth, the patient has survived for 28 years following her initial resection with surgical interventions.

Combined Modality Therapy↗

PubMed1990

Correlation between histologic grade of malignancy and copy number of c-erbB-2 gene in breast carcinoma. A retrospective analysis of 176 cases.

A system of histologic grade of malignancy in human breast carcinoma was devised by significantly modifying the way of evaluating number of mitoses and architectural atypia in the histologic grading of Bloom and Richardson. The modified grading system was applicable to all histologic subtypes of adenocarcinoma and showed a good association with prognosis of breast carcinoma patients in retrospective analysis of 176 consecutive surgical cases (P less than 0.0001). Of the three components of histologic grade, architectural atypia and number of mitotic figures independently had a significant effect on the prognosis. The copy number of c-erbB-2, a prognostic factor independent of tumor size and nodal status, was strongly correlated with the histologic grade, number of mitotic figures, and degree of nuclear atypia (P less than 0.001, each). Cox's regression model analysis showed that nodal status and histologic grade were two determinants of prognosis, and the independent effect of c-erbB-2 amplification was absorbed within that of the histologic grade. Although the importance of c-erbB-2 gene copy number seemed to be inferior to that of the histologic grade, both were shown to be strongly associated with the aggressiveness of the tumor itself rather than the extent of tumor spread.

Breast Neoplasms↗

PubMed1990

Difference in the ability of blood group-specific lectins and monoclonal antibodies to recognize the ABH antigens in human tissues.

Twelve different kinds of blood group-specific lectins have been used along with monoclonal anti-A, -B and -H antibodies for detecting the corresponding antigens in selected human tissues. Although most of the lectins recognized the antigens in the tissue sections examined, they displayed marked differences in their recognition patterns in certain tissues. Helix asparsa agglutinin (HAA), Helix pomatia agglutinin (HPA) and monoclonal anti-A antibody recognized A antigens in the mucous cells of salivary glands from blood group A or AB nonsecretor as well as secretor individuals, whereas Dolichos biflorus agglutinin (DBA), Griffonia simplicifolia agglutinin-I (GSA-I), Sophora japonica agglutinin (SJA) and Vicia villosa agglutinin (VVA) did not bind to them from nonsecretors. A antigens in endothelial cells, lateral membrane of pancreatic acinar cells and small mucouslike cells of submandibular glands from some individuals were likewise recognized by HAA and HPA but not by other blood group A-specific lections. In contrast, both HAA and HPA did not recognize the A antigens in mucous cells of Brunner's glands while other A-specific lectins and monoclonal anti-A antibody reacted specifically with the antigens. Such a difference was not observed with lectins specific for blood group B. However, the B antigens in Brunner's glands were recognized by these lectins but not with monoclonal anti-B antibody. The difference in labelling ability was also noted among the blood group H-specific lectins and monoclonal anti-H antibody in endothelial cells of blood vessels. Ulex europaeus agglutinin-I reacted with these cells irrespective of ABO and the secretor status of the individuals, while Anguilla anguilla agglutinin and monoclonal anti-H antibody reacted only with those cells from blood group O individuals. No reaction was observed with Lotus tetragonolobus agglutinin in these tissue sites. These results suggest a great diversity of blood group antigens in different human tissues.

ABO Blood-Group System↗

PubMed1990

Relationship between lectin binding properties and the expression of blood group ABH antigens in vascular endothelia and red blood cells from 18 primate species.

The reactivity was examined of horseradish peroxidase labelled Ulex europaeus agglutinin-I (UEA-I) and Griffonia simplicifolia agglutinin I-B4 (GSAI-B4) with red blood cells and vascular endothelium in formalin-fixed, paraffin embedded tissues from 18 primate species. The expression of blood group ABH antigens in these cells as well as secretions from other tissues was also examined by the indirect immunoperoxidase method using monoclonal anti-ABH antibodies as primary antibodies. In Prosimians and New World monkeys which lack ABH antigens on both red blood cells and endothelial cells, but produce these antigens in other tissue secretions, GSAI-B4 always reacted with both red blood cells and endothelial cells. In Old World monkeys, which express blood group antigens on endothelial cells but not on red blood cells, neither GSAI-B4 nor UEA-I reactivity were observed, except the endothelial cells from blood group B or O individuals occasionally reacted with GSAI-B4 or UEA-I, respectively. Although UEA-I reactivity was not observed in the endothelial cells of gibbon, it reacted with these cells from chimpanzees. In these two anthropoid apes, both endothelial cells and red blood cells expressed ABH antigens as in humans. These results suggest the close evolutionary relationship between the expression of blood group ABH antigens and lectin binding properties of red blood cells and endothelial cells in primate species.

ABO Blood-Group System↗

PubMed1990

Immunohistochemical study on overexpression of c-erbB-2 protein in human breast cancer: its correlation with gene amplification and long-term survival of patients.

Using a polyclonal antibody monospecific to the c-erbB-2 oncogene product, an immunohistochemical study on the expression of c-erbB-2 protein was performed in formalin-fixed, paraffin-embedded tissue sections from 176 primary breast carcinomas in which amplification of the c-erbB-2 gene had been detected in 28 cases. Expression of the c-erbB-2 protein was detected in 44 cases (25%), being strongly positive in 27 (15%) and weakly positive in 17 (10%). All cases with amplification of c-erbB-2 showed positive staining of its protein. There were only four cases in which c-erbB-2 was strongly expressed without amplification of the gene. In the group showing strongly positive staining, both overall and disease-free survival were significantly poorer than in the remainder of the cases. Using Cox's regression model analysis, overexpression of c-erbB-2 protein was demonstrated to be an effective prognostic factor independent of nodal status or tumor size.

Amino Acid Sequence↗

PubMed1990

Alveolar soft part sarcoma. A clinicopathologic and immunohistochemical study of 12 cases.

Twelve cases of alveolar soft part sarcoma (ASPS) were reviewed. Seven of them arose primarily in the lower extremities, three in the head and neck region, and two in other parts. ASPSs in the head and neck region occurred in children before 10 years of age, whereas ASPSs in the other regions occurred in rather older patients. Moreover, ASPSs of the head and neck were relatively small in size, and were diagnosed earlier than those in other regions. Histologically, six cases (including all the head and neck cases) contained considerable area of small and indistinct alveolar structures. Four cases showed remarkable cellular pleomorphism. Immunohistochemical demonstration of vimentin, desmin, the beta-subunit of enolase and the MM isozyme of creatine kinase, together with the absence of immunoreactive cytokeratin, supported the myogenic nature of this rare tumor. A small number of S-100 protein-positive tumor cells were also observed. Follow-up data for these cases disclosed that the tumors containing considerable area of small alveoli and uniform small tumor cells formed distant metastases at an early stage.

Adolescent↗

PubMed1990

Histochemical localization and analysis of blood group-related antigens in human pancreas using immunostaining with monoclonal antibodies and exoglycosidase digestion.

We examined the distribution of blood group-related antigens using an indirect immunoperoxidase method with monoclonal antibodies (MAb) directed to A, B, H, Lewis a (Lea), Lewis b (Leb), Lewis x (Lex), and Lewis y (Ley) antigens and Type 1 precursor chain in human pancreas. Effects of prior digestion with exoglycosidases on MAb stainings were simultaneously investigated. A, B, H, Leb, and Ley antigens were detected in acinar cells and interlobular duct cells but not in centroacinar cells, intercalated duct cells, and islet of Langerhans cells. The expression of these antigens in acinar cells was not dependent on Lewis type and secretor status of the tissue donors, whereas that in interlobular duct cells was strictly dependent on secretor status. The distribution pattern of these antigens in acinar cells was not homogeneous, i.e., cells producing H antigens expressed both Leb and Ley antigens but not A or B antigens, whereas those producing A or B antigens did not secrete Leb and Ley as well as H antigens. Digestion with alpha-N-acetylgalactosaminidase or alpha-galactosidase resulted in the appearance of Leb and Ley antigens as well as H antigen in acinar cells producing A and/or B antigens. Type 1 precursor chain was not detected in pancreatic tissues from secretors but appeared in acinar cells producing H antigen after alpha-L-fucosidase digestion, which also disclosed Lex but not Lea antigen in acinar cells expressing both Leb and Ley. In some non-secretors, MAb against Type 1 precursor chain reacted with acinar cells without enzyme digestion. Although Lea antigen was not detected in acinar cells, it was found in centroacinar cells, intercalated duct cells, and interlobular duct cells from all individuals examined except two Le(a-b-) secretors. After sialidase digestion, Lex antigen appeared in centroacinar and intercalated duct cells from some individuals. Sialidase digestion also elicited reactivity with MAb against Type 1 precursor chain in islet of Langerhans cells from some individuals. These results demonstrate the complexity in the pattern of expression and regulation of blood group-related antigens in different cell types of human pancreas. Such complexity may largely be ascribed to differences in individual genotypes and in gene expression patterns of different cell types.

ABO Blood-Group System↗

PubMed1990

[Basic study on the chemosensitivity test by miniaturized improved nucleic acid precursor incorporation assay (MINI assay)].

UNLABELLED: Miniaturized improved nucleic acid precursor incorporation assay (MINI assay) has been developed by Kern D. H. and Tanigawa N. et al since 1985. We investigated in vitro effects of mitomycin C (MMC), cisplatin (CDDP) and bleomycin (BLM) against A 549 lung cancer cells and HeLa cells by MINI assay. RESULTS: 3H-thymidine (3H-TdR) uptake in the positive control was 734.2 cpm +/- 10 cpm. The cut-off level for in vitro sensitivity was defined as more than 80% inhibition of 3H-TdR uptake in the drug treated cells compared to the positive controls. A 549 cells were sensitive to MMC (81.6%), CDDP (78.6%), but not to BLM (-57%). HeLa cells were sensitive to MMC (91%), CDDP (79.4%), but not to BLM (43%). MINI assay could be done with fewer cells than other methods and the results were obtained within 5 days. This method was considered to be useful for the chemosensitivity test with human tumors.

Antineoplastic Agents↗

PubMed1989

Correlation between long-term survival in breast cancer patients and amplification of two putative oncogene-coamplification units: hst-1/int-2 and c-erbB-2/ear-1.

The incidence and association with 10-year survival of amplification in five protooncogenes or transforming genes were retrospectively examined using DNAs extracted from formalin-fixed, paraffin-embedded blocks of tissues obtained from 176 consecutive patients surgically treated for primary breast carcinoma. The incidences of greater than threefold amplification of hst-1, int-2, c-erbB-2, ear-1 (one of c-erbA), and c-myc were 12, 13, 16, 10, and 4.0%, respectively. hst-1 and int-2 were almost always coamplified (21/22), while c-erbB-2 and ear-1 were frequently coamplified (18/28) with almost the same copy number. The hst-1 and int-2 pair and the c-erbB-2 and ear-1 pair, localized on chromosomes 11q13 and 17q21-22, respectively, in normal cells, were inferred to be constituents of different amplification units. Amplification of hst-1 and/or int-2 was detected preferentially in the younger age group, and was correlated with poorer prognosis in cases carrying four or more copies of the genes. Amplification of c-erbB-2 and/or ear-1 was strongly correlated with poor prognosis in all 176 patients, especially those with lymph node metastasis. Amplification of c-myc was also correlated with poor prognosis. Cox's life-table regression analysis showed that amplification of c-erbB-2 had a prognostic value, which was independent of other known prognostic factors such as lymph node status and tumor size.

Breast Neoplasms↗

PubMed1989

Histochemical demonstration of O-glycosidically linked, type 3 based ABH antigens in human pancreas using lectin staining and glycosidase digestion procedures.

Histochemical analyses of the chemical structures of sugar sequences with or without blood group specificity were carried out by combined stepwise digestion of tissue sections with exo- and endoglycosidases and subsequent lectin stainings in formalin-fixed, paraffin-embedded human pancreas. In acinar cells from blood group A or AB secretor individuals, sequential digestion with alpha-N-acetylgalactosaminidase and alpha-L-fucosidase imparted reactivity with peanut agglutinin (PNA) in cells reactive with Dolichos biflorus agglutinin as well as those with Ulex europaeus agglutinin I(UEA-I). Simple fucosidase digestion imparted the PNA reactivity only in UEA-I reactive cells. Sequential digestion with alpha-galactosidase and fucosidase likewise liberated the PNA binding sites in Griffonia simplicifolia agglutinin I-B4 reactive cells from blood group B and AB secretors. Sialidase digestion liberated the PNA binding sites not only in acinar cells but also intercalated duct cells, islet cells of Langerhans and endothelial cells. The PNA reactivity obtained by these enzyme digestions was eliminted by endo-alpha-N-acetylgalactosaminidase (endo-GalNAcdase) digestion. Preexisting PNA affinity in acinar cells from non-secretors was also susceptible to endo-GalNAcdase treatment. Following the endo-GalNAcdase digestion, fucosidase or sialidase digestion recovered the PNA reactivity in acinar cells from nonsecretors. These results show that ABH determinants carried on O-glycosidically linked type 3 chain (D-galactose-(beta 1-3)-N-acetyl-D-galactosamine alpha 1-serine or threonine) are secreted in pancreatic acinar cells and suggest that product coded by the secretor gene is required for the complete conversion of type 3 precursor chains into H determinants.

ABO Blood-Group System↗

PubMed1989

Micro extraction of DNA from whole blood and amniocytes.

We describe methods for extracting genomic DNA from a small amount of whole blood or cultured amniocytes. Nuclear DNA was extracted from whole blood spotted on blotting paper. Relatively large molecules of DNA with the average amount of 7-9 micrograms was extracted from 1 ml of blood spotted and stored for at most two years, being roughly 1/3 of that extracted directly from fresh whole blood. The estimated minimum amount of whole blood that gives a suitable autoradiogram of Southern hybridization was 0.3 ml. Another series of amounts of whole blood or an amniocyte suspension were molded in low-melting agarose into an 100 microliter gel block. The DNA extracted from a block that was made from at least 0.25 ml of whole blood, or from 1.25 x 10(5) amniocytes (equivalent to 1/8 of the number of confluent cells in a 25 cm2 culture flask) resulted in one suitable Southern analysis. Both methods described here are applicable to the diagnosis of newborns and/or fetuses at risk of a genetic disease and to the diagnosis of a patient from whom a large amount of blood material is difficult to obtain. These methods also make a long-way transportation of the materials possible.

Adult↗