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

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

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

[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↗

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

Chromophobe renal cell carcinoma: a report of two cases.

Chromophobe renal cell carcinoma (RCC) is a recently established subtype of RCC, which has rarely been reported in Japan. In this communication, the authors report two Japanese cases of chromophobe RCC together with the immunohistochemical findings. The tumors were composed of sheets and cribriform glands formed by tumor cells with cloudy and reticular cytoplasm. Ultrastructurally, the cytoplasm was filled with numerous microvesicles. The tumor cells were positive for cytokeratin, epithelial membrane antigen, and Tamm-Horsfall protein. Occasionally, LeuM1-positive cells were also noted. Vimentin was negative, unlike the usual RCC. Reactivity for peanut agglutinin was more frequent than that to Lotus tetragonolobus agglutinin. The results of this study suggest that the tumor cells possessed phenotypes similar to the distal nephron rather than to the proximal tubular cells.

Adult↗

PubMed1992

Isolation of subclones with different tumorigenicity and metastatic ability from rat nephroblastoma cell line, ENUT.

Seven subclones derived from a chemically induced rat nephroblastoma cell line, ENUT, were isolated and their cytological characteristics were examined in order to investigate the biological nature of the parent tumor. The subclones were divisible into two types based on morphological and biological features: polygonal cells with well developed junctional complexes and less developed cell surface microvilli, showing a doubling time of 13.0 to 14.0 h and tumorigenicity in nude mice of 42-100%, and spindle-shaped cells with poorly developed junctional complexes and well developed microvilli, showing a doubling time of 9.7 to 12.2 h and 100% tumorigenicity. Potential for both spontaneous and experimental metastasis to the lungs was apparently higher in the spindle-shaped clones than in the polygonal clones. In spite of their differing characteristics, the cytochemical phenotypes of the two clonal types were almost identical to those of immature renal tubules of the developing fetal kidney. The present findings suggest that these subclones possess characteristics of early nephrogenic epithelial cells, and we speculate that the biological differences observed among the subclones may be explained by differences in malignant progression. In addition, these clones appear to be a useful model for the study of tumorigenicity and metastasis.

Animals↗

PubMed1992

Alpha-fetoprotein-producing immature mediastinal teratoma showing rapid and massive recurrent growth in an adult.

A case of immature mediastinal teratoma in a 43-year-old Japanese man is reported. The tumor, which was multicystic with solid zones and measured 12 x 6 x 8 cm, arose in the anterior mediastinum. The serum alpha-fetoprotein (AFP) level was elevated to 5,114 ng/ml before surgery. Histologically, the solid zones showed an admixture of irregular glands lined by columnar or cuboidal epithelium set in a spindle cell stroma, some foci of primitive neural tissue, and scattered small nests of hepatoid cells. Immunohistochemically, the hepatoid cells and epithelia lining some of the cysts showed a strongly positive reaction for AFP. Eight months after surgery, the patient died of respiratory failure caused by a rapidly growing massive recurrent tumor, which measured 40 x 24 x 13 cm, in the left thoracic cavity. However, the elevated serum AFP level had been decreasing during the course of the recurrence in response to chemotherapy. The recurrent tumor showed remarkable proliferation of loose mesenchymal tissue without primitive neural tissue. These findings suggest that immature mediastinal teratoma in adults is highly malignant, and that non-AFP-producing mesenchymal tissue played a critical role in forming the rapidly growing massive recurrent tumor in the present case.

Adult↗

PubMed1992

Glomerular lesions in unilateral nephrectomized and diabetic (UN-D) mice.

Experimental diabetes was induced in both control and unilaterally nephrectomized male mice by injecting streptozotocin (SZ) (50 mg/kg x 5 days) one week after nephrectomy. The time course changes in the glomerular lesions were examined for up to 12 weeks after completion of the SZ-injection (12WAI). In unilateral nephrectomized and diabetic mice, mild segmental expansion of the mesangial area developed at 4WAI, and it progressed to prominent segmental glomerulosclerosis at 12WAI. In the electron microscopic examination at 12WAI, marked expansion of the mesangial area, segmental thickening of the glomerular basement membrane, fusion of the foot processes of podocytes and a prominent increase in the number of microvilli of capillary endothelial cells were observed. On the other hand, mild to moderate expansion of the glomerular mesangial area was only sporadically found in unnephrectomized diabetic mice at 12WAI. Interestingly, Bowman's capsules of diabetic mice were generally lined with flattened epithelia but those of non-diabetic mice with cuboidal or low columnar epithelia.

Animals↗

PubMed1992

Characterization of experimental rat nephroblastoma and its cell line.

Rat nephroblastoma (Wilms' tumor) was induced by transplacental administration of N-ethyl-nitrosourea (ENU). The induced renal tumors were histologically compatible with human nephroblastoma. A cultured cell line (ENU-T-1) established from a xenotransplant, showed similar morphological and biological features to cultured embryonal kidney cells. Introduction of normal human chromosome #11 (#11) bearing Wilms' tumor suppressor gene(s) (WT) suppressed colony-forming ability on soft agar plates (CFA) but tumorigenicity of ENU-T-1 was not affected. Whereas tumorigenicity of human nephroblastoma cell line, SK-NEP-1 was completely suppressed, CFA was unchanged. These facts indicated that pathogenetic mechanism is different between human and experimental rat nephroblastomas.

Animals↗

PubMed1992

[A 13-week percutaneous toxicity study of prednisolone farnesylate (PNF) gel in beagle dogs with a recovery period of 5 weeks].

The toxicity of Prednisolone farnesylate (PNF) gel, a synthetic glucocorticoid, was investigated in the Beagle dog. PNF gel was administered percutaneously at doses of 0.2, 0.8 and 3.2 mg/kg/day for 13 weeks, then the drug was withdrawn for 5 weeks to evaluate the reversibility. In addition, 2 mg/kg/day prednisolone gel (PN gel), which is approximate to 3.2 mg/kg/day PNF gel in prednisolone molarity, was also administered for comparison. The results are summarized as follows: 1. No deaths were observed in any of the PNF gel test groups or the PN gel group, nor were there any abnormal findings in the clinical signs of the animals. 2. In the hematology, a tendency toward a decrease in the lymphocyte ratio was observed in males from the PNF gel 0.8 mg/kg/day and above groups. In the PN gel group, a significant decrease or a tendency toward a decrease in the lymphocyte ratio was observed, as well as an increase in the white blood cell count in some animals. 3. In the blood biochemistry, a significant decrease or a tendency toward a decrease in total cholesterol and phospholipid was observed in males from the PNF gel 3.2 mg/kg/day group and a tendency toward an increase in triglyceride in females from the PNF gel 3.2 mg/kg/day group was observed. In the PN gel group, a tendency toward an increase in AIP activity, a tendency toward an increase in triglyceride were observed. 4. In the histological examinations, a decrease in the weight of the thymus and adrenal glands, vacuolation of hepatocytes in the middle zone of the liver, atrophy of zona fasciculata of the adrenal glands, hypertrophy of zona glomeruli, swelling of cortical cells of zona faciculate and atrophy of the thymus were observed in the PNF gel 0.8 mg/kg/day and above groups. In the PN gel group, atrophy of submandibular lymph nodes and mesenteric lymph nodes was observed in addition to the same changes as observed in the PNF gel groups. Furthermore, thinning, atrophy or a decrease in the weight of the adrenal glands was also observed both in the PNF gel 3.2 mg/kg/day group and the PN gel group at the end of the 5-week recovery period. 5. As described above, a decrease in the lymphocyte ratio, in the weight of the thymus and adrenal glands and vacuolation of hepatocytes were observed in the PNF gel 0.8 mg/kg/day and above groups.(ABSTRACT TRUNCATED AT 400 WORDS)

Administration, Cutaneous↗

PubMed1992

[A 52-week percutaneous toxicity study of prednisolone farnesylate (PNF) gel in beagle dogs with a recovery period of 8 weeks].

The toxicity of Prednisolone farnesylate (PNF) gel, a synthetic glucocorticoid, was investigated in the Beagle dog. PNF gel was administered percutaneously at doses of 0.05, 0.2 and 0.8 mg/kg/day for 52 weeks, then the drug was with held for 8 weeks to evaluate reversibility. The results are summarized in the following. 1. In the 0.05 mg/kg/day and above groups, hypotrichosis in the application site of the skin, thinning of the skin and atrophy of the appendages, and in the 0.2 mg/kg/day and above groups a tendency toward retarded body weight gain, were observed. 2. In the 0.2 mg/kg/day and above groups, a drop in the lymphocyte ratio, a rise in GOT activity and A1P level, and in the 0.8 mg/kg/day group a rise in free fatty acid were observed. 3. In the 0.2 mg/kg/day group and above groups, atrophy of the zona fasciculata and zona reticularis were observed. In the 0.8 mg/kg/day group, a decrease in the weight of the thymus and adrenal glands, and a increase in the weight of the liver, were observed. 4. At the end of the recovery period, most of the changes disappeared, except for those in the adrenals and treated area. From the above results, under the conditions of this study, it was concluded that when the changes observed in the application site of the skin in each group were not taken into consideration, the toxicological no effect level was 0.05 mg/kg/day for both males and females and the overt toxic dose level was 0.8 mg/kg/day.

Administration, Cutaneous↗

PubMed1991

Two cases of a renal epithelial tumour resembling immature nephron.

Two cases of renal epithelial tumours are reported in females aged 46 and 66 years respectively. In spite of the large size of the tumours, neither invasive growth nor metastasis was observed. Histologically, the tumours were composed of immature epithelial cells forming tubules with abortive glomeruloid structures. Electron microscopy of tumour cells revealed poorly developed polarity and intracytoplasmic organelles. They showed similar immunohistochemical reactions to those of developing nephrons, particularly to those of the S-shaped body. The nuclear DNA content of the tumour cells was almost euploid. We conclude that the lesions were epithelial tumours of the kidney histologically mimicking developing renal parenchyma.

Aged↗