Biomedical subjects

Y Nagashima

Publications and source records attributed to Y Nagashima.

At least 19 recordsLinked to original sources

PubMed1995

Trypsinogen expression in human ovarian carcinomas.

Increased secretion of matrix metalloproteinases and serine proteinases is well known to be associated with cancer invasion and metastasis. We aimed to elucidate the implication of trypsin, a serine proteinase and a representative digestive enzyme in invasion and metastasis of human carcinomas. Northern blot, RT-PCR and Western blot analyses and immunohistochemical studies were performed to detect and analyze trypsinogen expression in 5 ovarian carcinoma cell lines and 10 human ovarian carcinoma tissues using a DNA probe for trypsinogen I, and monoclonal and polyclonal antibodies to human trypsin I. Among the 5 ovarian carcinoma cell lines, only the MCAS (mucinous cystadenocarcinoma) cell line showed a high level of trypsinogen production and mRNA expression by Western and Northern blot analyses, respectively. However, Southern blot analysis of RT-PCR products could detect considerable levels of trypsinogen mRNA in all ovarian cancer cell lines. In Northern analysis of ovarian cancer tissues, all advanced cancer samples showed trypsinogen gene expression. Serous cystadenocarcinomas exhibited particularly high levels of gene expression. Immunohistochemical staining also detected trypsin in ovarian carcinoma tissues. In contrast, normal ovaries and tumors with low malignant potential did not show trypsinogen expression. Our results demonstrate the extra-pancreatic production and distribution of trypsinogen in human ovarian carcinomas.

Adenocarcinoma↗

PubMed1995

A novel insertion sequence (IS)-like element of the thermophilic bacterium PS3 promotes expression of the alanine carrier protein-encoding gene.

A novel insertion sequence (IS)-like element was found in the 5'-upstream region of the alanine carrier protein-encoding gene (acp) in the thermophilic bacterium PS3 chromosomal DNA. The sequence contained an open reading frame (ORF) encoding a polypeptide of 369 amino acids which revealed high similarity with ORFs from IS891 from the cyanobacterium Anabaena and IS1136 from Saccharopolyspora erythraea. The direction of transcription was the same as that of acp, and typical inverted and direct repeats characteristic of IS were found in both the 5' and 3' region of the ORF. Southern hybridization analysis of the chromosomal DNA revealed that multiple copies of the ORF sequence were contained in the PS3 genome. This element might well be a member of a new IS family including IS891 and IS1136, and we have designated this element IS1341. The analysis of acp expression in Escherichia coli cells indicated that IS1341 promotes the expression of acp.

Amino Acid Sequence↗

PubMed1995

Establishment and characterization of two human ovarian clear cell adenocarcinoma lines from metastatic lesions with different properties.

Two permanent human ovarian clear cell adenocarcinoma lines (OVISE and OVTOKO) were established from metastatic tumors of two patients who were treated with five to six courses of CAP chemotherapy. The two cell lines grow on monolayers and showed a variety in both size and shape: small or moderately sized cuboidal cells, columnar cells, spindle-shaped cells, and malignant tumor giant cells. The cell lines have been in culture for 4 to 6 years, the passage number varying from 160 to 220. The mean population-doubling time of the two cells was 60 to 70 hr. The OVISE cells shed tumor-associated antigens CA19-9, CA125, and TPA in the culture medium, whereas the OVTOKO cells did not secrete them at detectable levels. Immunohistochemical analysis showed that coexpression of cytokeratins and vimentin was preserved in the two cell lines, which is a feature of cultured epithelial origin. Cytokeratin polypeptides 7, 8, 18, and 19 were expressed in both cell lines. The EGF receptor was more intensely expressed in the OVTOKO cells than in the OVISE cells. The estrogen and progesterone receptors were negative in both cell lines. The two cell lines showed no chemosensitivity to anticancer drugs including cisplatin, doxorubicin, cyclophosphamide, and etoposide. Heterotransplantation of the two cell lines reflected the origin of cells. Intraperitoneal transplantation of the OVTOKO cells yielded peritoneal implantation and distant metastasis, whereas that of the OVISE cells showed no dissemination and metastasis. These new ovarian clear cell adenocarcinoma lines will provide a relevant experimental system for further investigations into the intrinsic alterations responsible for malignant progression and chemoresistance.

Adenocarcinoma, Clear Cell↗

PubMed1995

Marked induction of gelatinases, especially type B, in host fibroblasts by human ovarian cancer cells in athymic mice.

Two human ovarian adenocarcinoma cell lines, MCAS-3 and OVISE-3 were found to secrete little of any type of gelatinase in tissue culture. However, when these cell lines were implanted subcutaneously into nude mice the cyst fluids from the resultant tumors contained gelatinase A and/or B. The enzyme activities, especially of gelatinase B, were much higher in the malignant MCAS-3 tumors than in those of the less malignant OVISE-3 tumor cells. To elucidate the origin of gelatinase B in cyst fluids of the MCAS-3 tumors, murine skin fibroblasts (MSF) were isolated from a subcutaneous tumor in a nude mouse and tested for their proteinase secretion in culture. MSF cells, which secreted some gelatinase A and gelatinase B, were induced to secrete high levels of both enzymes, especially gelatinase B, by co-cultivation with MCAS-3 cells. In addition, gelatinase A activity was induced by incubation of MSF cells with the conditioned medium of either MCAS-3 or OVISE-3 cells, whereas gelatinase B was induced only with that of MCAS-3. Although cytokines or growth factors such as IL-1 beta, TGF-beta 1, TNF-alpha or EGF stimulated the secretion of gelatinases A and B from MSF cells, their effects on gelatinase B activity were far less than that of the MCAS-3 conditioned medium. These results indicate that the major part of gelatinase B activity in the cyst fluids of the ovarian tumors is secreted by host interstitial cells stimulated by tumor-derived humoral factors. Similar tumor cell-host cell interactions may be important in the production of various proteinases in other tumor types.

Animals↗

PubMed1995

Detection of a proteinaceous toxin in the brackishwater clam (Corbicula japonica).

Water extracts from the brackishwater clam (Corbicula japonica) were found to be lethal to mice upon i.v. injection. Muscular tissues (foot muscle, adductor muscle, mantle muscle, mantle and siphon) were all toxic while gill and viscera (mid-gut gland, testis and ovary) were nontoxic; the highest toxicity was observed with foot muscle. The toxin was judged to be a thermolabile, basic protein with a mol. wt. of about 13,000.

Animals↗

PubMed1995

Establishment of two rat osteosarcoma cell lines (YROS-1 and YROS-2) induced by radioactive phosphorus.

Two rat osteosarcoma cell lines, YROS-1 and YROS-2, were established from two experimental osteosarcomas and induced by internal irradiation with radioactive phosphorus. Both cell lines formed a monolayer cell sheet in vitro with focal piling. The YROS-1 cells were refractile and spindle or polygonal in shape, whereas the YROS-2 cells were flat, spread and polygonal in shape. Ultrastructurally, the YROS-1 cells had well-developed rough-surfaced endoplasmic reticulum with focal pericellular deposition of calcified matrix, whereas YROS-2 had abundant polysomes and intracytoplasmic filaments. Both cell lines grew stably with population doubling times of 23 and 39 h, respectively. Flow cytometry revealed that YROS-1 was rich in proliferating cells compared to YROS-2, with a higher colony-forming efficiency. YROS-1 showed high alkaline phosphatase activity, while YROS-2 possessed low activity. When subcutaneously transplanted into lumbodorsal area of athymic nude mice, only YROS-1 formed tumors with frequent lung metastasis.

Animals↗

PubMed1995

[Clinico-pathological aspect of HIV-sero positive inpatients].

To describe the clinico-pathological aspect of HIV/AIDS inpatients, we reviewed HIV/AIDS inpatients in our hospital since 1988. We evaluated risk factors, CD4 counts, CD4/8 ratio, cause of death and the survival time after diagnosis of AIDS. A total of 13 HIV sero-positive subjects were admitted to our hospital, including 11 cases (85%) with AIDS. One patient was a foreigner. All cases were male; age range 18-70 years. Hemophiliacs and sexually transmitted patients account for 62% and 38% of cases, respectively. There were no cases of intravenous drug users. In all cases with AIDS, CD4+ cells and CD4/8 ratio significantly decreased with median value of 73/microns and 0.18, respectively. Six cases (54%) were decreased and autopsy were done in four cases. Findings of autopsy revealed HIV encephalopathy and systemic various infection, including Pneumocystis carinii, cytomegalovirus, papilloma virus and pox virus. Cytomegalovirus infection was found in all cases; one had innumerable inclusion bodies in lung, adrenal gland and intestines. The median survival time after diagnosis of AIDS were 26 months.

AIDS-Related Opportunistic Infections↗

PubMed1995

Localization and activity of adrenal carbonyl reductase during the estrous cycle.

Changes in carbonyl reductase (CR) activity towards seven substrates in adrenal glands of female rats were investigated during estrous cycle and the circadian patterns of adrenal CR activity towards seven substrates in male rats were examined. The localization of the enzyme protein in the adrenal gland was carried out by immunohistochemistry. In female rats, adrenal CR activity was lowest between 1200 and 2000 h on the day of proestrus, the phase in which a large, significant increase in ovarian steroids, gonadotropins and prolactin is observed. Immunoreactivity in the zona reticularis of the adrenal cortex to anti-adrenal CR, CR-K2, antibody on the day of proestrus was faint or negative compared with that on other days of the estrous cycle. In male rats, no major changes in adrenal CR activity were observed compared with females, although these were slight changes in all the enzyme activities. These results suggest that adrenal CR may be negatively regulated by the pituitary-adrenal axis in female rats and, in particular, on the day of proestrus.

Adrenal Glands↗

PubMed1995

Deodorization of laboratory animal facilities by ozone.

Deodorizing effect of ozone was investigated comparing two types of compact ozonizing apparatus made on an experimental basis. The concentrations of ammonia and trimethylamine were examined as an indicator for deodorizing effect of ozone in animal rooms of rats and guinea pigs at laboratory animal facilities of three different universities. Both of the ozonizing apparatus were able to remove ammonia and trimethylamine in animal rooms, with no significant difference in the performance of the two apparatus.

Ammonia↗

PubMed1995

Effects of fasting and refeeding on some metabolic characteristics of rat brown adipose tissue.

To investigate the effects of fasting and refeeding on rat interscapular brown adipose tissue (BAT), we determined in vitro oxygen consumption, in vitro lipolysis, DNA content and composition of fatty acid (FA). Fasting decreased the weights of body and BAT, and the DNA content of BAT. Fasting reduced both basal oxygen consumption and basal lipolysis of BAT. Fasting also suppressed the noradrenaline- and glucagon-induced increases in both oxygen consumption and lipolysis of BAT. In both triglyceride and phospholipid FA of BAT the extent of unsaturation was increased by fasting. Refeeding restored all of these parameters to the control level. These observations indicate that fasting reduces both oxygen consumption and lipolysis of BAT, resulting in reduced thermogenesis, while fasting could stimulate BAT in certain phases of its function as observed in FA composition.

Adipose Tissue, Brown↗

PubMed1995

Immunocytochemical localization of estrogen and progesterone receptors (ER & PgR) in breast cancer.

The localization of estrogen receptors (ER) and progesterone receptors (PgR) in human breast cancer was examined light and electron microscopically by using specific monoclonal antibodies. Among the breast cancer patients, ER-positive (ER[+]) cases were shown by enzyme immunoassay (EIA) to account for 58.9% (63/107), and 63.5% of these cases were women in the postmenopausal state. The PgR-positive (PgR[+]) rate was 50.5% (54/107), with 44.4% of these positive cases being postmenopausal women. The rates of ER(+) and PgR(+), ER negative (ER[-]) and PgR negative (PgR[-]), and ER(+) and PgR(-) cases were 43.9% (47/107), 34.6% (37/107) and 15.0% (16/107), respectively. Although ER(-) and PgR(+) cases were in few number, they were found (6.5%, 7/107). These results correlated well with those obtained by the immunocytochemical method. In either case, i.e anti-ER or anti-PgR reaction, positive nuclei and negative nuclei were found intermingled with each other in a given visual field. In electron microscopy, both anti-ER and anti-PgR antibodies bound to sites in the euchromatin area of the nucleus.

Antibodies, Monoclonal↗

PubMed1995

[Squamous cell carcinoma of the lung with high CT number, but without calcification].

Squamous cell carcinoma of the lung was diagnosed in a 64-year-old man. Chest roentgenographic and tomographic examinations did not show areas of high-density in the tumor, but chest CT scan showed diffuse, scattered, high-density nodules in the tumor. Histologic examination did not show calcification, and calcium staining (Kossa method) was negative. Examination of the thin-slice CT, the soft X-ray film, and the resected slice of the lung indicated that the high-density nodules seen in the tumor on the chest CT scan corresponded to markedly fibrotic lesions with severe anthracosis. This case shows that high-density areas on a CT scan may reflect non-calcified lesions.

Calcinosis↗

PubMed1995

[Pulmonary malignant lymphoma associated with bilateral diffuse linear, reticular, and consolidated shadows].

A 72-year-old asymptomatic man was admitted to our hospital with abnormal chest radiographic findings: bilateral diffuse linear, reticular, and consolidated shadows. Malignant lymphoma (diffuse small-cell B-cell lymphoma) was diagnosed histologically after an open lung biopsy and was confirmed by a genetic examination. The lymphoma cells had infiltrated widely in various regions of the lung, including the alveolar spaces and walls, the bronchial and vascular spaces, and the pleura. This diffusion of the lymphoma cells might have accounted for the chest X-ray findings. Malignant pulmonary lymphoma should be included in the differential diagnosis patients with diffuse linear, reticular, and consolidated shadows on chest X-ray films.

Aged↗

PubMed1995

[A case of thymic carcinoma successfully controlled by combined chemotherapy and irradiation].

There is no well established therapeutic protocol for advanced thymic carcinoma, although chemotherapy and irradiation can apparently be used to control the malignancy. Here we report a case of advanced thymic carcinoma successfully controlled by combination of treatment with systemic chemotherapy and irradiation. A 79-year-old man was admitted to the Hiratsuka Kyosai Hospital with complaints of productive cough and dyspnea. Chest roentgenogram and computerized tomography on admission revealed a tumorous lesion in the anterior mediastinum and on accumulation of left pleural and pericardial effusion fluid. The tumor was diagnosed as a thymic squamous cell carcinoma, after percutaneous needle biopsy. Because of extension to the left pleura and cardiac sac, the tumor was considered to be unresectable and systemic chemotherapy (CDDP + VDS + MMC) was begun. After drainage of the pericardial effusion fluid, intrapericardial injection of MMC was also done. The anterior mediastinum was irradiated. The patient has been doing well, with regression of the thymic tumor for 48 months after irradiation. This case provides important information that can be used to establish an effective therapeutic regimen for advanced thymic carcinoma.

Aged↗

PubMed1994

Identification of one- and two-chain forms of trypsinogen 1 produced by a human gastric adenocarcinoma cell line.

It has previously been reported that two kinds of human gastric adenocarcinoma cell lines (STKM-1 and MKN28) secrete a trypsin-like enzyme. In this study, four molecular forms of the enzyme (26, 25, 24 and 23 kDa on non-reducing SDS/PAGE) were purified from the serum-free conditioned medium of STKM-1 cells. Analysis of N-terminal amino acid sequences showed that the 26 kDa protein was a two-chain form of trypsinogen 1 which had been produced by proteolytic cleavage of the Arg107-Val108 bond of trypsinogen 1, and the 24 kDa protein was the one-chain form of trypsinogen 1. The 25 and 23 kDa proteins were the activated forms of the two-chain and one-chain trypsinogen 1 respectively. Isoelectric focusing gave pI values of 6.3 and 6.6 for the 26 kDa two-chain form and the 24 kDa one-chain form of trypsinogen 1 respectively. Comparison of the proteolytic activities indicated that the one-chain trypsin 1 had amidolytic activity about four times higher than that of the two-chain enzyme.

Adenocarcinoma↗

PubMed1994

Frequent somatic mutations and loss of heterozygosity of the von Hippel-Lindau tumor suppressor gene in primary human renal cell carcinomas.

We analyzed 47 primary sporadic human renal cell carcinomas (39 clear cell and 8 non-clear cell) for mutations of the von Hippel-Lindau (VHL) tumor suppressor gene using the polymerase chain reaction and single strand conformational polymorphism analysis of DNA. All of the positive cases in single strand conformational polymorphism analyses were further characterized by direct sequencing. Somatic mutations were detected in 22 (56%) of 39 clear cell renal carcinomas including 15 deletions, 3 insertions, 3 missense mutations, and 1 nonsense mutation. Nineteen of these mutations predicted to produce truncation of the VHL protein. These mutations mainly occurred in the last one-third region of exons 1, 2, and 3. In addition, loss of heterozygosity of the VHL gene was observed in 16 (84%) of 19 informative clear cell renal carcinomas. No somatic mutations were detected in 8 non-clear cell carcinomas. These results show that the VHL tumor suppressor gene is one of the major tumor suppressor genes in human renal cell carcinomas, especially in the clear cell subtype renal cell carcinoma. Clear cell carcinoma might be distinguished from other pathological types of renal cell carcinomas by molecular genetic techniques.

Base Sequence↗

PubMed1994

Establishment and characterization of a malignant melanoma cell line (YP-MEL) derived from a patient with neurocutaneous melanosis.

A cell line, YP-MEL, was established from an intracranial malignant melanoma occurring in a neurocutaneous melanosis (NCMsis) patient. The established cell line was successfully cultured in serum-free medium with a doubling time of 41 h. The cells were refractile and small in size, with occasional pigmented giant cells. Histochemical and immunohistochemical features were compatible with common malignant melanoma and its cell line. Chromosome analysis revealed many supernumerary chromosomes and marker chromosomes including double minutes (DMs). When transplanted into nude mice, YP-MEL formed tumors histologically consistent with the original tumor. Addition of sera to the medium caused cellular spreading and elongation of cytoplasmic processes with an increase of melanin contents and tyrosinase activity. Because there was no melanoma cell line derived from a NCMsis patient, YP-MEL might be a beneficial tool for study on NCMsis.

Biopsy↗

PubMed1994

Establishment of three rat soft tissue tumor cell lines with different degrees of myogenic differentiation.

Soft tissue malignancies often show divergent differentiation, including myogenic lineage. Five rat tumors induced with 20-hydroxymethylcholanthrene (20-OH-MCA), were cultured in vitro, and three cell lines (YMC-1, YMC-2, YMC-3) were established from them. YMC-2 and -3 cells were spindle-shaped, and YMS-1 cells round and epithelioid. In confluency, YMC-3 cells formed myotubes. The nude mouse xenotransplants showed morphological features consistent with their myogenic phenotypes. Muscle-specific enzyme activities were highest in YMC-3 cells. These lines may be useful in the investigation of the myogenic differentiation of undifferentiated mesenchymal cells.

Animals↗