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Biomedical subjects

T Torigoe

Publications and source records attributed to T Torigoe.

At least 73 records · Page 4Linked to original sources

Development of monoclonal antibodies reacting against mycobacterial 65 kDa heat shock protein by using recombinant truncated products.

A mycobacterial 65 kDa molecule is a member of the GroEL heat shock protein family. We developed mAbs reacting against recombinant 65 kDa protein by using a gene (pTB12) which encodes this protein. Three mAbs (B20, B97 and B167) reacted selectively with 65 kDa proteins of Mycobacterium tuberculosis, BCG and Mycobacterium leprae, although B20 and B167 may weakly react with a 15 kDa molecule of mammalian cells. One (B108) was obviously cross-reactive between mycobacterial 65 kDa and the mammalian intracytoplasmic protein. We also developed deletion mutants of pTB12. The localization of these mAb-defined epitopes was determined by using truncated proteins of the Mycobacterium tuberculosis 65 kDa molecule produced in E. coli. Immunohistochemical analysis showed that B20, B97 and B167 mAbs could detect this antigen in experimental granulomas induced by injection of BCG in the subcutaneous tissue of rats. These mAbs should be useful for analyzing the immunobiologic roles of mycobacterial 65 kDa molecules.

Animals↗

[A case of benign mixed tumor of the trachea].

A very rare case of benign mixed tumor of the trachea was reported. A 52-year-old male was admitted to our hospital because of hemoptysis, slight dyspnea and stridor. Bronchoscopic examination revealed a polypoid tumor which arose from the anterior wall of the upper trachea, obstructing about 70% of the tracheal lumen. For the purpose of getting pathological specimen and securing the air way in anesthesia, endoscopic polypectomy (2/3 of the tumor) was done by using GIF-XP 20. Circumferential resection of the trachea (4 rings) with end-to-end anastomosis was performed one month after polypectomy. To our knowledge, this was the 7th reported case in Japan. Clinical studies and operative procedures of this disease were briefly discussed.

Humans↗

Heat- or stress-inducible transformation-associated cell surface antigen on the activated H-ras oncogene-transfected rat fibroblast.

A WKA rat fetus fibroblast (WFB) was transfected by several oncogenes including EJras (activated H-ras), polyoma middle T (PyMT), v-src, c-myc, and adenovirus type 12 E1A-E1B. We analyzed the expression of the transformation-associated cell surface antigens on WFB by developing monoclonal antibodies. One of the WFB transformation-associated cell surface antigens, recognized by monoclonal antibody 067, was constitutively expressed only on two (W31 and W70) of ten WFB-EJras-transformed clones. This antigen could not be detected on parental WFB cells as well as ten WFB-PyMT transformant clones. Furthermore, it was not expressed on several clones of partially transformed WFB-v-src and WFB-adeno E1 transfectants or nontransformed WFB-c-myc transfectants. Monoclonal antibody 067 could form an immunoprecipitate with an approximate molecular weight of 67,000 which was composed of a single polypeptide chain. It was also shown that the expression of this antigen could be enhanced by cyclic AMP or cholera toxin treatment of W31; treated cells also showed a phenotypic reversion to the nonmalignant growth characteristics of the parental WFB. Moreover, the expression of this antigen could be induced on the WFB-EJras transformants such as W14, which do not constitutively express this antigen, by treatment of these agents. Furthermore, the expression of antigen was enhanced by heat and superoxide treatment on W31. These data suggest that the monoclonal antibody 067-defined molecule is a novel transformation-associated cell surface antigen that could be induced by heat shock or other physiological stress.

Animals↗

[Fetal screening of congenital heart disease].

Echocardiograms of two hundred and ninety normal fetuses at 18-39 weeks of gestation were studied with a real-time linear scanner (3.5 MHz). Aortic valvular diameter, pulmonary valvular diameter, mitral valvular diameter, tricuspid valvular diameter and total cardiac dimension were measured. These parameters increased with fetal growth. The ratios of pulmonary to aortic valvular diameter and of tricuspid to mitral valvular diameter were more than 1, suggesting right ventricular dominance in the fetal hearts. The cardio-thoracic ratio was about 50% in this period. The four-chamber view, left ventricular long-axis view, ventricular short-axis view, short-axis view of the great arteries, the view of the aortic arch and the view of the inferior and superior vena cava were also recorded. These six views could be obtained in 9.8% of all subjects at the same time. Detection rates for these views were the greatest in pregnancy at 22-33 (especially 28-29) weeks of gestation. The ventricular short-axis view and short-axis view of the great arteries were easily detected in the dorsoposterior position, whereas the view of the aortic arch was obtained in the dorsoanterior position. It was concluded that fetal echocardiography needed to be performed at least twice, at 22-23 and 28-29 weeks of gestation, and this would be useful in diagnosing some congenital heart diseases.

Aorta, Thoracic↗

[Clinical studies of long-term administration of OK-432 in patients with gynecologic cancer].

The clinical value of OK-432 treatments, was evaluated in 215 patients with various types of gynecologic cancer. Five KE OK-432 was administrated by intramuscular injection every other day for up to 5 years, unless severe side-effects were observed. The 5-year survival rate was determined in 51 patients in the OK-432 group and 93 patients in the control group. The survival rate in the OK-432 group was significantly higher in the cervical cancer patients in stages III and IV than in the control group, at from 20 to 29 months and from 37 to 42 months. These results indicated that long period administration of OK-432 might improve the survival rate in patients with gynecologic cancer, especially in those with advanced cervical cancer.

Adult↗

[Expression of cell surface transformation-associated antigens TAAs on activated oncogene-transfected rat fibroblast and antitumor immunity by hosts].

The key point in this study was that the cell surface antigens in the events of transformation were analyzed using the same clonal-derived nontransformed cell and various oncogenes. WKA rat fetus-derived fibroblast line, WFB, showed the strict non-transformant phenotypes. Transfection of EJras and polyoma middle T oncogenes into WFB resulted in the acquisition of tumorigenicity in vitro and in vivo. The newly expressed cell-surface antigens on some transformants were analyzed using four monoclonal antibodies (Moabs) (#109, #061, #081 and #067) that were produced after immunization of BALB/c mice with EJ-ras-transformed WFB clone, W31 and W14. #109 recognized a glycoprotein composed of a single polypeptide chain with 86 KD. This antigen was also expressed in the FJ ras transformants of Fisher rat fetus-derived 3Y1 fibroblast, FTL43 rat T lymphoma, SP56 rat gliosarcoma, and ca. 16th-day embryo. #061 could react with WFB transformants, but not with fetus. Furthermore, these two antigens are expressed on TGF-beta-stimulated WFB. However, #109 but not #061 is expressed on PDGF-stimulated WFB, indicating that #109 expression does not require the transforming process. It is also indicated that these two antigens act as target structures in the Natural killer (NK) cytotoxicity assay. #081 could react with WFB transformants but not with FTL43, SP56 and 3Y1 ras transformants, suggesting a limited expression of #081 defined antigen on WFB transformants. #067 reacted with W31 but not with W14. This antigen is expressed on 5 clones out of approximately 40 highly tumorigenic WFB ras transformed clones. We also analyzed the expression dynamics of these TAAs during the morphological reversion of W31 to parental WFB using cAMP analogues such as dibutylic cAMP. The data indicated that, although W31 cells were reversed morphologically to WFB, the level of #109 antigen expression was not changed. #061 and #081 defined antigens were reduced by cAMP treatment for 4-5 days. However, it is of interest that #067-defined antigen expression was greatly enhanced during the morphological reversion of W31 to WFB by cAMP. This was also true when W31 was treated with 1% FCS for 5 days and heat shock, suggesting that this antigen is a heat shock-related protein.

Animals↗

[Results of abdominal surgery in patients over 90 years old].

During a year period (1978-1987), 105 cases of abdominal surgery were performed at Sasebo Chuo Hospital on patients over 80 years of age. This study discusses 4 patients who were over 90 years old at time of their surgery. Emergency surgery had been required for 2 patients (a 91-year-old male with a sigmoid colon ca., and a 91-year-old female with a descending colon ca.), and elective surgery had been performed on the other two patients (a 95-year-old male with a caecum ca., and a 97-year-old female with a leiomyosarcoma of stomach). The patients who underwent elective surgery had a uniformly smoother hospital course and an overall better prognosis then the patients who underwent emergency surgery, whose prognosis was poor. We have concluded that early diagnosis and elective surgery is the necessary procedure in the elderly patient.

Adenocarcinoma↗

[Regulation of progesterone accumulation in dissociated luteal cells on day 15 of pregnancy in rats].

Luteal cells from the corpora lutea of rats on day 15 of pregnancy were suspended in DMEM-F12 and progesterone accumulation was measured by radioimmunoassay. Progesterone accumulation increased steadily over eight hours and then gradually decreased. Progesterone accumulation was seen to increase with the addition of BSA and 25-hydroxycholesterol. At doses of 12 ng/ml and 600 ng/ml hCG, progesterone accumulation increased, but was seen to decrease at a dose of 30 micrograms/ml hCG. However, the addition of cyclohexamide alone had no effect on progesterone accumulation, but the simultaneous addition of cyclohexamide and hCG blocked the effects of hCG in a dose related manner. These data indicate that progesterone production by rat luteal cells depends on cholesterol availability. The results reveal that hCG stimulates progesterone production by a process requiring new protein synthesis.

Animals↗

[Morphological and functional changes in 3 beta-HSD positive cells in corpora lutea during pregnancy in rats].

On days 5, 10, 15 and 20 of pregnancy, rat corpora lutea (CLs) were dissected and dissociated into single cell suspensions by enzyme treatments. To assess 3 beta-hydroxysteroid dehydrogenase (HSD) activity, a histochemical suspension-staining procedure was used. The number of cells positive for 3 beta-HSD in CL were 148.0 +/- 13.7, 130.1 +/- 25.4, 134.0 +/- 23.5 and 116.8 +/- 13.5 X 10(3) cells on days 5, 10, 15 and 20 of pregnancy, respectively. The 3 beta-HSD positive cells increased in size from 18.5 +/- 0.28 microns on day 5 to 35.7 +/- 0.50 microns on day 20 of pregnancy. The suspended luteal cells were incubated in serum-free DME-F12 for 20 hours with or without human chorionic gonadotropin (hCG, 100 ng/ml) or dibutyryl cyclic AMP (dbcAMP, 10mM) to test their functionality, and progesterone accumulation was determined by radioimmunoassay. Progesterone secretion from the 3 beta-HSD positive cells was maintained at the same levels until day 15 and decreased significantly on day 20 of pregnancy. However the response to hCG stimulation of the 3 beta-HSD positive cells decreased significantly on day 20, the 3 beta-HSD positive cells maintained the same responsiveness to dbcAMP stimulation on progesterone secretion throughout pregnancy. These data suggest that the steroidogenic rat luteal cells may be regulated by morphologically and functionally different mechanisms, and that progesterone may be secreted by at least two different pathways.

3-Hydroxysteroid Dehydrogenases↗

Characterization of n-butyl alcohol solubilized, breast tumor specific antigens recognized by a human autologous cytotoxic T-cell clone.

We demonstrated previously the establishment of a human cytotoxic T-cell clone, TcHMC-1, under culturing with recombinant interleukin that showed the specific cytotoxicity against an autologous breast tumor cell line, HMC-1-8. In the present study, the autologous tumor specific antigens that could be involved in this cytotoxicity were extracted by using n-butyl alcohol and were analyzed for their biochemical profiles. The cytotoxicity of TcHMC-1 against HMC-1-8 was inhibited by adding OKT3 and OKT8 monoclonal antibodies into the cultures, or by pre-sensitizing HMC-1-8 target cells by anti-major histocompatibility complex class I monoclonal antibodies. This suggests that T-cell antigen receptor molecule complexes Ti/T3 on TcHMC-1 and corresponding specific tumor antigens on HMC-1-8 are involved in the cytotoxicity under the restriction of major histocompatibility complex class I products. Precultures of TcHMC-1 with crude n-butyl alcohol extracts from HMC-1-8 cells enhanced the cytotoxic potentials of this clone as seen as mixed lymphocyte tumor cell cultures. This enhancement was dependent on dosage of crude n-butyl alcohol extracts and these TcHMC-1 cells were still cytotoxic specifically for HMC-1-8 targets, but not for other allogenic tumor lines including K562. However, HMC-1-8 crude n-butyl alcohol extracts could not enhance DNA synthesis of TcHMC-1 as assessed by incorporation of [3H]thymidine in the cells. Biochemical purification studies demonstrated that the HMC-1-8 tumor specific antigens were eluted into fractions containing molecules with molecular weights of approximately 200,000 on Sephadex G-200 column chromatography. The antigens were further separated into the fraction that was eluted with 0.4-0.5 M NaCl in an ionic strength on Mono Q fast protein liquid chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of this fraction demonstrated three molecules with molecular weights of 26,000, 30,000, and 32,000 under reduced molecular conditions. The data suggest that these molecules could be tumor specific antigens that are involved in the cytotoxicity of cytotoxic T-cells against a human autologous tumor.

1-Butanol↗

Identification of the transformation-associated cell surface antigen expressed on the rat fetus-derived fibroblast.

A WKA rat fetus-derived fibroblast cell line WFB showed strict nontransformant phenotypes in vitro such as anchorage dependency of cell growth in soft agar, contact inhibition, and serum dependency on the monolayer cell culture. Transfection of 6.6-kilobase EJras oncogene into WFB resulted in the acquisition of tumorigenicity in vitro and in vivo. The cell surface antigen that is moderately or highly expressed on these WFB transformants, designated as W14 and W31, was analyzed using monoclonal antibody 109 that was produced after the immunization of BALB/c mice with W31. Moab 109 recognized a glycoprotein with a molecular weight of 36,000 composed of a single polypeptide chain with 5.4 isoelectric point value. This antigen was highly expressed on WFB EJras and polyoma middle T-DNA transformants, but was undetectable or at the best only faintly recognized on WFB parental cells, transfectants of WFB with c-myc, and normal thymus, liver and kidney of WKA adult rats. It was also clearly expressed on the EJras transformants of Fisher rat fetus-derived 3Y1 fibroblast, but very faintly on parental 3Y1. Furthermore, this antigen was detected on some rat T-lymphoma and gliosarcoma lines. However, it was undetectable on EJras transformants on NRK-49F rat kidney cells and NIH3T3 and BALB3T3 mouse cells. In addition, this antigen appeared on the cell surface of concanavalin A-activated WKA rat lymphocytes and WKA rat on the 16th day of embryo but not on the 8th. These results suggested that the cell surface antigen detected by Moab 109 was clearly unrelated to the ras oncogene product p21 that was highly expressed on EJras-transformants of WFB or 3Y1 cells. Furthermore, it was shown that W14 and W31 cells but not parental WFB cells were susceptible to rat splenic NK cells that were induced by poly(I-C) treatment. Pretreatment of these W14 or W31 cells with Moab 109 could block the NK cell activity against W14 and W31. These data suggest that this antigen may act as one of the NK target structures, and plays an important role as a tumor antigen on the host tumor surveillance, since the antigen was expressed (a) on the cell surface after the cell transformation or enhanced DNA synthesis of some particular cells, and (b) in the W31 tumor developing progressively in the syngeneic rats.

Animals↗

Structure and complete nucleotide sequence of the gene encoding chicken cytosolic adenylate kinase.

The gene encoding cytosolic adenylate kinase (AK1) was isolated from a chicken genomic DNA library by using its cDNA as a hybridization probe. The chicken AK1 gene spanned about 6 kilobase pairs and consisted of 7 exons. Analyses of the 5'-flanking region sequence and S1 nuclease mapping revealed that the transcription initiation site is located at 84 base pairs upstream from the ATG initiation codon. The TATA box and the putative CAT box were located 29 base pairs and 97 base pairs upstream from the transcription initiation site, respectively. A total of 7 GC boxes were found in the 5'-flanking region, the exon 1, and the intron 1. The GC boxes were surrounded by the sequences with extremely high G+C contents. When projected on the three-dimensional structure of the AK1 protein molecule, introns fell either between or near the ends of alpha-helices and beta-strands, and most of the coding exons encoded at least one alpha-helix and one beta-strand. The dot matrix plot analysis between chicken AK1 and bovine mitochondrial adenylate kinase (AK2) suggested that the AK1 gene might have evolved from the AK2 gene by deletion of one or more exon(s).

Adenylate Kinase↗

The expression of tumor rejection antigen on rat fetus fibroblasts transformed by the ras oncogene.

The expression of tumor rejection antigens (TRA) was analyzed on clones of rat fetus-derived fibroblasts, WFB, transformed or transfected by oncogenes. It was shown that a tumorigenic W14 clone, which is an activated H-ras transformant of parental WFB, expressed TRA in transplantation experiments using syngeneic WKA rats. The data also showed that W14 TRA was acquired in the event of cell transformation, since it was not detected on parental non-transformed WFB cells or Wmyc-4 clone which is a transfectant of WFB by mouse plasmacytoma-derived c-myc DNA. However, TRA was not expressed or at least was not detected on highly tumorigenic W31, another clone of H-ras transformants of parental WFB, in the transplantation experiments. We also assessed the level of expression of major histocompatibility antigens (MHC) class I molecules on these cells by using R4-8B1 Mab that reacts specifically with rat class I antigen. The data indicated that it was decreased on W14, W31, and Wmyc-4, but not on parental WFB. Although this molecule was weakly positive on W14 cells, W31 and Wmyc-4 showed even greater decreases. These data may indicate that the TRA expression and its recognition by syngeneic hosts are dependent upon the transformed clones, although their parental cell is the same. We discuss in detail this difference of expression and recognition of TRA in the context of the cell transforming process.

Animals↗

Inhibitory effects of adrenocorticotrophin or corticosterone on progesterone secretion during mid-pregnancy in rats.

Conceptus number was reduced to one on Day 7 of pregnancy in rats by aspirating all but a single conceptus (Group E) or left at greater than or equal to 8 conceptuses (Group C). In Group E rats, serum progesterone concentrations remained low from Day 12 until autopsy at Day 21. Hypophysectomy on Day 12 significantly increased serum progesterone values after Day 17 of pregnancy, and these increases were blocked by treatment with ACTH (10 U/day, i.p., Days 12-17). Adrenalectomy on Day 12 also induced slight, but statistically significant, increases in serum progesterone concentration after Day 17, and these were overcome by implantation of a 10 mg capsule of corticosterone. In Group C rats, hypophysectomy or adrenalectomy on Day 12 did not change serum progesterone concentrations, but 40 U ACTH/day inhibited progesterone secretion. We conclude from these results that the pituitary-adrenal system exerts inhibitory effects on progesterone secretion during mid-pregnancy in rats.

Adrenocorticotropic Hormone↗

Characterization of tumor rejection antigen molecules of chemically induced murine colon tumor C-C26.

The molecular nature of a tumor-specific transplantation antigen (TSTA) of a chemically induced BALB/c mouse colon tumor C-C26 was investigated. The antigen was noncytolytically extracted by 2.5% n-butanol treatment of the cells. Crude butanol extract from C-C26, but not from colon tumor C-C51 and fibrosarcoma Meth-A of BALB/c mice could provide protection against the challenged C-C26 tumor in the transplantation experiment. Crude butanol extract from another syngeneic colon tumor C-C36 also induced a cross-protection against the challenged C-C26 tumor. C-C26 crude butanol extract was characterized by biochemical procedures including the Sephadex G200 column, lens culinaris affinity column, and anion-exchange Mono Q fast protein liquid chromatography column, and by the enzyme digestion study of the antigens and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The data indicated that C-C26 TSTA was eluted into fractions containing molecules of approximately Mr 200,000 on Sephadex G200 column chromatography. This antigen was also found in unbound fractions on a lens culinaris affinity column. The antigen was further separated into the fraction that was eluted with 0.4 M NaCl in an ionic strength on Mono Q fast protein liquid chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of this fraction showed the molecule with a molecular weight of 30,000. The enzyme digestion study indicated that the immunogenicity of the antigen was inactivated by papain but probably not by neuraminidase treatment. These data suggest that the immunogenic moiety of C-C26 TSTA molecules is located in the peptide portions rather than in sialic acid residues or carbohydrate portions. Furthermore, there are several similarities of the molecular characteristics between C-C26 TSTA and previously reported C-C36 TSTA, such as the amenability to n-butanol extraction. Lens culinaris lectin inaffinity, and ionic strength.

Animals↗

Use of bromocriptine in the treatment of normoprolactinemic infertility.

The efficacy of bromocriptine therapy was studied in 84 normoprolactinemic infertile patients. Bromocriptine (2.5-5.0 mg/day) was given for at least 1 month. Bromocriptine therapy was effective in 54 of 84 cases (64%), including 6 of 15 (40%) cases with amenorrhea, 16 of 24 cases (67%) with anovulatory cycle, 9 of 15 cases (60%) with delayed ovulation and 23 of 30 cases (77%) with luteal phase defect. Sixteen cases were able to conceive with bromocriptine alone. Twenty-five patients who did not respond to bromocriptine were treated with a combination therapy consisting of bromocriptine and clomiphene. Of the 25 cases, 14 responded to the therapy, and 6 of them were able to conceive. The response of prolactin to domperidone (10 mg, i.v.) was significantly (p less than 0.01) higher in the group responding to bromocriptine than in the nonresponding group. These results indicate that the administration of bromocriptine is an effective therapy for patients with normoprolactinemic endocrine disorders, and that domperidone may be useful in selecting the candidates for bromocriptine therapy.

Adult↗

Heterogeneous distribution of acidic TA-4 in cervical squamous cell carcinoma: immunohistochemical demonstration with monoclonal antibodies.

Tumor antigen TA-4 is divided into two subgroups; acidic and neutral TA-4. The tissue localizations of these TA-4 subgroups were examined by using monoclonal antibodies, i.e., Mab-21 which reacts with both acidic and neutral TA-4, and Mab-317 which is specific to acidic TA-4. Immunohistochemical staining with Mab-21 showed positive cells in most parts of the cancer nest and in the intermediate layer of the non-cancerous squamous epithelium of the uterine cervix, whereas positive staining with Mab-317 was observed only in the cells at the peripheral parts of the cancer nest adjacent to the surrounding stromal tissue. Thus, examination of the subgroups of TA-4 may be a useful aid for investigating the biologic behavior of squamous cells.

Antibodies, Monoclonal↗

Flow cytometric analysis of tumor antigen TA-4 in cervical cytologic specimens.

Flow cytometric analysis of cellular contents of TA-4, a tumor antigen of squamous cell carcinoma, was performed in vaginal smears from 143 women. Abnormal findings, characterized by a cell peak with high fluorescence intensity, appeared in 85.7% of cases (54/63) of cervical squamous cell carcinoma, 80% of cases (4/5) of severe dysplasia, 42.9% of cases (3/7) of mild-to-moderate dysplasia, and in 20.6% of cases of healthy women (13/63). Cellular compositions sorted from abnormal cell fractions with high fluorescence intensity consisted, not only of malignant cells but also of morphologically nonmalignant cells. Immunohistochemical studies indicated that these nonmalignant cells with high TA-4 content originated from the intermediate layer of squamous epithelium adjacent to the cancer nest. These findings indicate that this method may be a useful approach for investigating some biologic characteristics of cells and for detecting cervical cancer or its precursors.

Antigens, Neoplasm↗