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Biomedical subjects

K Washiyama

Publications and source records attributed to K Washiyama.

At least 37 records · Page 2Linked to original sources
PubMed1993

Expression of vascular endothelial growth factor in capillary hemangioblastoma.

The gene expression of vascular endothelial growth factor (VEGF), an endothelial cell-specific mitogen, was examined in capillary hemangioblastomas which are composed of abundant microvasculature and primitive angiogenic elements represented by stromal cells. Northern blot and in situ hybridization analyses showed that the VEGF mRNA was highly expressed in the stromal cells, but not in the endothelial cells. Reverse transcription/PCR analysis revealed that the VEGF mRNA species expressed in the stromal cells were the shorter splicing forms, VEGF121 and VEGF165, which are potent to be secreted from cells and the longer splicing form, VEGF189, which is predominantly cell-associated. Our findings suggest that VEGF secreted from the stromal cells plays an important role in the endothelial cell proliferation in capillary hemangioblastomas.

Base Sequence↗
PubMed1992

Human glial fibrillary acidic protein (GFAP): molecular cloning of the complete cDNA sequence and chromosomal localization (chromosome 17) of the GFAP gene.

We isolated three glial fibrillary acidic protein (GFAP) cDNA clones from a glioma cell line, U-251 MG. One clone isolated from a U-251 MG cDNA library was long, but lacked both ends. Using poly(A)+ RNA and primers synthesized according to the sequence of this clone, we used the polymerase chain reaction-assisted rapid amplification of cDNA ends (PCR-RACE) method, which is a strategy to isolate cDNA ends, and obtained cDNA clones for the 5' and 3' ends. From the sequences of these overlapping clones, the complete nucleotide sequence of human GFAP cDNA was established. The start (ATG) and the stop (TGA) signals were seen at nucleotide positions 15 and 1311, respectively, and divided the entire sequence of 3027 bp into 14 bp of 5' non-coding, 1296 bp of coding and 1717 bp of 3' non-coding regions. Using cDNA probes made from both the coding and the 3' non-coding regions, Northern blot hybridization was performed with two different stringencies on RNAs from human and rodent brains and human GFAP-positive and -negative cells. It was shown that the 3' non-coding region probe was more specific for human GFAP than the coding region probe which was specific only under higher stringency conditions. This was also suggested by homology analysis of the sequence with those of various intermediate filament proteins. Based on these findings, we performed spot blot hybridization of sorted human chromosomes and Southern blot hybridization of PCR-amplified DNAs of a panel of hamster-human somatic cell hybrids and localized the human GFAP gene to chromosome 17.

Amino Acid Sequence↗
PubMed1992

Gynecomastia and ectopic human chorionic gonadotropin production by transitional cell carcinoma of the bladder.

We report a patient with gynecomastia and ectopic production of human chorionic gonadotropin (HCG) by a transitional cell carcinoma of the bladder. In the present case, serum HCG levels and gynecomastia paralleled the clinical course. On admission, the patient was suffering from invasive transitional cell carcinoma (grade 3) of the bladder with metastasis to the left inguinal lymph nodes, together with gynecomastia. The serum HCG level was also elevated. After anticancer chemotherapy, the apparent bladder lesion and gynecomastia disappeared, and the serum HCG level declined to within normal limits. About 2 months after discharge, when the patient suffered from recurrent invasive tumors of the bladder, gynecomastia reappeared and the serum HCG level again became elevated. beta-HCG was demonstrated in biopsy tissue using the immunoperoxidase technique. The presence of beta-HCG was always focally demonstrated and was shown to be localized in the cytoplasm of the tumor cells.

Carcinoma, Transitional Cell↗
PubMed1991

Supralevator pelvic exenteration with colonic J-pouch-anal anastomosis and Mainz pouch operation with anastomosis to the urethra. Report of a case.

This is the first documented case report in which construction of double pouches, a colonic J-pouch and a Mainz pouch with anastomosis to the urethra, was performed in a patient who underwent supralevator pelvic exenteration. We have shown that the operation is feasible; the patient was continent, except for nightly urinary leakage, and was able to defecate and urinate spontaneously without any catheterization. Based on the experience of this case, we believe that this combined operation results in a better quality of life for the patient because of the avoidance of both urinary and fecal stomas. If justified pathologically, this operation may represent an improvement for those patients with extensive cancer of the rectum, which invades the adjacent anterior viscera, or that of the bladder, which invades the posterior viscera.

Adult↗
PubMed1989

Primary malignant lymphoma of the brain: demonstration of immunoglobulin gene rearrangements in four cases by the Southern blot hybridization technique.

Using the Southern blot hybridization technique, four cases of the primary malignant lymphomas of the brain, histologically diffuse large cell lymphoma, were examined for the immunoglobulin gene rearrangements. In three lymphomas, the rearrangements were observed in both heavy and light chain genes, providing strong evidence for a B cell lineage of these tumors. On the other hand, in the remaining lymphoma, the rearrangement was observed only in the heavy chain gene. Despite this, immunohistochemical examination revealed positive stainings for heavy and light chain immunoglobulins in tumor cells, suggesting the occurrence of light chain gene rearrangement at the undetectable level. Thus, B lymphocytic differentiation at the gene level was demonstrated in three, or possibly all, of the primary intracerebral malignant lymphomas examined. Since no more than two rearrangements were detected in each immunoglobulin gene, these lymphomas were considered to be monoclonal in nature.

Adult↗
PubMed1989

Differential expression of glial fibrillary acidic protein in human glioma cell lines.

We have obtained a cDNA fragment to human glial fibrillary acidic protein (GFAP) by immunoscreening a lambda gt11 human brain cDNA library with antibody to bovine GFAP. The highly homologous nucleotide sequence of this clone with that of the mouse GFAP enabled the identification of this cDNA as one encoding GFAP. As this cDNA hybridized with a single major RNA species in Northern blots of RNA from human and mouse brain tissues and gave one or two bands in Southern blots of human genomic DNA, it was considered to be specific for GFAP. Using this cDNA as a probe we investigated the levels of GFAP expression in ten human glioma cell lines. A 3.5-kb GFAP mRNA was detected in five of the ten glioma cell lines, one of which was U-251 MG cell line and the other four were clones derived from the same tumor (CL1, 2, 3, and 4). There was a difference in the amount of GFAP mRNA among U-251 MG and the four clonal cell lines. Quantitative evaluation of this difference by RNA dot blot analysis revealed that the amount of GFAP mRNA expressed in CL3 was about 1/5 and in CL4 about 1/10 the amount expressed in U-251 MG, CL1, and CL2. Semiquantitative Western blot analysis showed that GFAP levels corresponded to the GFAP mRNA levels in these cell lines. By Southern blot analysis of genomic DNA the GFAP gene was similarly detected in all of these cell lines regardless of the level of GFAP expression.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1989

Neuroendocrine markers in central nervous system neuronal tumors (gangliocytoma and ganglioglioma).

We studied five cases of central nervous system neuronal tumor, one gangliocytoma and four gangliogliomas, both ultrastructurally and immunohistochemically, using antibodies to neuroendocrine markers including tyrosine hydroxylase (TH), serotonin (5HT), somatostatin (SOM), met-enkephalin (MEK), leu-enkephalin (LEK), substance P (SP), gastrin, vasopressin, oxytocin, vasoactive intestinal polypeptide, adrenocorticotropic hormone and calcitonin. In all cases, the presence of dense-core vesicles (60-250 nm) in the neuronal elements was the characteristic ultrastructural finding. Synapses were observed in two cases. Immunohistochemically, variable numbers of neuronal cells showed positive staining for SOM in five cases, TH, MEK and LEK in three cases, and 5HT and SP in one case each. The others were negative. Positive immunoreactivity for multiple markers was shown in all cases. SOM, TH, 5HT and SP were present in the small- to medium-sized cells, while MEK and LEK were almost exclusively confined to the large cells. Our study clearly indicated that these tumors contained neuronal cells which were not homogeneous with regard to neuroendocrine markers.

Biomarkers↗
PubMed1989

Tumor-infiltrating lymphocytes and histocompatibility antigens in primary intracranial germinomas.

Subpopulations of tumor-infiltrating lymphocytes (TIL's) and the major histocompatibility complex (MHC) antigens of neoplastic cells were examined in three intracranial germinomas by an immunohistochemical method using monoclonal antibodies. About 70% to 80% of TIL's were T lymphocytes which were either infiltrating diffusely or in clusters, whereas 20% to 30% of TIL's were B lymphocytes which tend to cluster in tumor tissues. Examination of T lymphocyte phenotypes revealed both the cytotoxic/suppressor and helper/inducer T lymphocytes, as in other tumors. However, the existence of a considerable number of B lymphocytes in the TIL population was uncommon and seemed to be a characteristic feature of the intracranial germinoma, which might suggest a difference of host immune response to this neoplasm as compared to other tumors. On examination of the MHC antigens, no MHC class I or II antigens in the neoplastic cells were stained, while positive staining for both antigens was seen in the TIL and stroma tissues. From these findings, it was suggested that the degree of TIL infiltration might not be correlated with the expression of MHC antigens in neoplastic cells in cases of primary intracranial germinoma.

Adolescent↗
PubMed1989

DNA synthesis, blood flow, and glucose utilization in experimental rat brain tumors.

The relationships between distribution of deoxyribonucleic acid (DNA)-synthesizing cells (S-phase cells) and blood flow and glucose utilization were investigated in rat brain tumors using an autoradiographic technique and immunoperoxidase staining for bromodeoxyuridine (BUdR). Two strains of rat brain tumor were used: strain A and B, both induced by the Rous sarcoma virus. Strain A was biologically more malignant than strain B. The blood flow was unevenly distributed in the tumor, compared with the contralateral cortex, the average blood flow in the tumor was about 50% in strain A and 60% in strain B. The distribution of blood flow did not correlate with the distribution of S-phase cells or with the distribution of vessels in the tumor in either strain A or B. The average glucose utilization in strain A was about 250% and in strain B about 170% of that of the contralateral cortex. The high glucose utilization area correlated well with the distribution of BUdR-positive nuclei in strain B. These findings suggest that the biological malignancy of a tumor correlates with glucose utilization rather than with blood flow, and that malignant brain tumors show a marked increase in glucose utilization for nucleic acid synthesis.

Animals↗
PubMed1988

Correlation of DNA ploidy and morphological features of human glioma cell cultures with the establishment of cell lines.

Eleven gliomas were serially cultivated and examined for DNA distribution by flow cytometry and simultaneously for morphological features by light microscopy at the various passage levels until passage 50 at most. Seven gliomas (four low-grade gliomas, three anaplastic gliomas) showed a similar DNA distribution pattern with a main diploid and small tetraploid peaks at various passages. In this group, only one culture formed a permanent cell line, whereas six cultures showed a limited growth ranging from 6 to 24 passages. In contrast, the other four gliomas (each an anaplastic glioma) showed a marked change of DNA distribution through passages and finally a single DNA aneuploid population prospered. Each of these four gliomas yielded established cell lines. Thus, it is suggested that the change of DNA ploidy and prosperity of DNA aneuploid populations in flow cytometry might be used as early and reliable indices for the later establishment of glioma-derived permanent cell lines. Since the changes of DNA distribution are frequently associated with the morphological changes, as seen in the latter group, careful tracing of morphological features is valuable in determining of the fate of cultures, especially in the absence of a flow cytometer. The correlation between the potential to become established cell lines and histology of the original gliomas is also discussed.

Adult↗
PubMed1988

An immunohistochemical study of cytokeratin and glial fibrillary acidic protein in choroid plexus papilloma.

Cellular localization of cytokeratin and glial fibrillary acidic protein (GFAP) was examined in two normal choroid plexuses and five choroid plexus papillomas by the peroxidase-antiperoxidase (PAP) method and double immunofluorescence (IFL) microscopy. Cytokeratin was observed in the majority of epithelial cells in all samples of normal and neoplastic choroid plexuses. On the other hand, GFAP was observed in some of the constituent epithelial cells in two cases of papilloma. Most of these GFAP-positive papilloma cells were simultaneously positive for cytokeratin, as could be seen by the PAP stainings of serial sections and by the double IFL stainings of the same sections. From these findings, it was suggested that normal and neoplastic choroid plexus epithelial cells usually express cytokeratin and that some of the neoplastic cells can simultaneously express both cytokeratin and GFAP.

Adult↗
PubMed1988

Immunohistochemical analysis of tumor-infiltrating lymphocytes and major histocompatibility antigens in human gliomas and metastatic brain tumors.

A subpopulation of tumor-infiltrating lymphocytes (TILs) and the expression of major histocompatibility complex (MHC) antigens of the tumor cells were examined in 14 glioma and 13 metastatic brain tumor tissues. In both tumors, most of the TILs were T lymphocytes, and both phenotypes of the cytotoxic/suppressor and helper/inducer T lymphocyte were found. On examination of MHC antigens, beta 2-microglobulin was shown intensely on tumor cells in all cases, and the monomorphic determinant of the human leukocyte antigen (HLA-DR) was shown in 10 glioma and in 5 metastatic cases. The correlation between the number of TILs and MHC antigen expression on tumor cells was equivocal as a whole in cases of both glioma and metastatic brain tumor.

Adolescent↗
PubMed1988

[Local immunotherapy with OK-432 for malignant gliomas--immunohistochemical analysis of chronological changes of tumor tissues].

Chronological changes of glioma tissues treated with local immunotherapy with OK-432 were examined by immunohistochemical method. OK-432 was injected into glioma tissues through Ommaya's reservoir 3 days (3 patients), 7 days (2 patients) and 14 days (2 patients) prior to the operation. Frozen sections surgically obtained from these patients were stained with avidin-biotin-peroxidase complex method using Leu-series monoclonal antibodies for pan T lymphocytes (Leu-1), cytotoxic/suppressor T lymphocytes (Leu-2 a), helper/inducer T lymphocytes (Leu-3 a), B lymphocytes (Leu-12), MHC class I antigen (beta 2m) and MHC class II antigen (HLA-DR). In 2 out of 7 glioma tissues obtained before local injection of OK-432, only few T lymphocytes were found infiltrating around the small blood vessels. In all glioma tissues obtained 3 and 7 days after injection, coagulation necrosis of glioma tissues was observed within 1-2 cm from Ommaya's tube and many T lymphocytes granulocytes and macrophages were infiltrating diffusely in the glioma tissues. Whereas in all glioma tissues obtained 14 days after injection, coagulation necrosis was also observed, however granulocytes and macrophages were scarce. The most of the infiltrating cells were T lymphocytes. Examination of T lymphocytes phenotypes revealed that both cytotoxic/suppressor and helper/inducer phenotypes of T lymphocytes were intermingled with each other in all cases. beta 2m was expressed on the most of glioma cells in all cases before and after injection. Whereas HLA-DR antigen was expressed on the tumor cells in 4 out of 7 cases before injection, however this antigen was expressed in all cases after injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1987

[Immunohistological analysis of infiltrating lymphocyte subpopulations in gliomas and metastatic brain tumors].

Subpopulations of infiltrating lymphocytes were studied by immunohistological method using monoclonal antibodies in gliomas and metastatic brain tumors. Thirteen specimens from 8 glioma patients, and 7 specimens from 3 metastatic brain tumor patients were used. No special therapy for brain tumor had been performed in these cases, but 3 glioma patients and all metastatic brain tumor patients had received steroid hormone. Frontal lobe obtained from the autopsy case of chest trauma was served as a normal control. Frozen sections were stained with avidin-biotin peroxidase complex method using Leu-series monoclonal antibodies for pan T-cells (Leu-1), cytotoxic/suppressor T-cells (Leu-2 a), helper/inducer T-cells (Leu-3 a) and B-cells (Leu-12). Lymphocyte infiltrates were quantitated by counting positively stained cells in 13 glioma and 7 metastatic brain tumor specimens. In normal frontal lobe, only a few T-cells infiltrated around several blood vessels in the parenchyma and subarachnoid space. But in the cases of glioma, many perivascular lymphocytic infiltrates were found and in the cases of metastatic brain tumor, many lymphocytes were found diffusely in the interstitial area between nests of tumor cells. Most of these lymphocytes were T-cells and B-cells were scarce, and Leu-2 a and Leu-3 a positive cells intermingled with each other. Len-3 a/2 a ratio ranged from 0.2 to 0.9 in the half of gliomas and 1.5 to 3.6 in another half of gliomas, three of which were treated with steroid hormone.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1986

Primary malignant lymphoma of the brain: an immunohistochemical study of eight cases using a panel of monoclonal and heterologous antibodies.

Frozen sections of eight primary malignant lymphomas of the brain were examined by the immunohistochemical methods using a panel of monoclonal and heterologous antibodies to B lymphocyte (immunoglobulins, BA-1, Leu-12 and HLA-DR), T lymphocyte (OKT-11 and Leu-1) and monocyte (OKM-1) surface markers. Paraffin sections were also used in the examination of cytoplasmic immunoglobulins. Surface and/or cytoplasmic immunoglobulins (Ig) were observed in seven cases, four of which were shown to be distinctly monoclonal and the other three less so. The remaining 1 case showed no distinct staining for Ig. BA-1, Leu-12 and HLA-DR stainings were positive in four, four and five cases, respectively. The marker phenotypes of (BA-1, Leu-12, HLA-DR) were shown to be (+, +, +) in one lymphoma, (+, -, -) in three, (-, +, +,) in three, and (-, -, +) in one. Thus, it was demonstrated that the present lymphoma cases showed a marked immunological heterogeneity, and it was shown that all of them including the Ig-negative case revealed one or more of these three additional B cell markers, indicating B cell lineage of these cases. Examination of T cell and monocyte markers revealed positive staining in normal or reactive lymphoid cells distributed around blood vessels or sporadically in tumor tissues, but not in lymphoma cells. Epstein-Barr virus (EBV)-associated nuclear antigen was not demonstrated in the seven cases examined, making it unlikely that these lymphomas were related with EBV infection.

Antigens, Neoplasm↗