Biomedical subjects

N Konishi

Publications and source records attributed to N Konishi.

At least 91 records · Page 5Linked to original sources

PubMed1998

Overexpression of CD44 variant transcripts in rat transplantable thyroid carcinoma lines demonstrating lung metastasis.

Expression of CD44 isoforms has been reported to be involved in tumor invasion and metastasis in both rodents and man. We earlier documented establishment of rat transplantable thyroid carcinoma lines in vivo from primary lesions induced by a chemical carcinogen. Recently, two lines (L1a-M4 and L2a-M6) were found to spontaneously metastasize to the lung after subcutaneous transplantation. To determine whether CD44 splice variants contribute to their metastatic spread, carcinoma lines with and without lung metastasis were evaluated quantitatively and qualitatively using RT-PCR followed by hybridization and immunohistochemical analyses. The L1a-M4 and L2a-M6 metastatic lines showed significant overexpression of CD44 variant transcripts containing variant exons v4-v6 or v9-v10/v8-v10, respectively, with concomitant reduced levels of standard transcripts. Investigation of the precise composition of alternatively spliced mRNA in normal tissues and carcinoma lines using an exon-specific RT-PCR method, revealed major chain variant transcripts containing v2/v3, v4-v6, v7-v10 and v8-v10 in all specimens. Applying the same RT-PCR analysis to mRNAs derived from cultured cell lines, demonstrated essentially the same pattern. The results suggest that quantitative increase rather than qualitative change in CD44 variant isoforms is associated with the pathogenesis of lung metastasis of rat thyroid carcinomas.

Alternative Splicing↗

PubMed1998

[A case of acute renal failure caused by rhabdomyolysis with thrombosis of the deep vein of the right leg, following sleeping in a straight sitting position for a long time].

A 41-year-old woman took an overdose of sedatives on the 13th of January, 1994 and remained a sleep in the straight sitting position until she was woken up on the 15th. The next day she consulted our hospital complaining of pain and swelling of her right leg. X-ray examination with contrast medium revealed obstruction of the deep vein of her right leg by a thrombus. On the 17th, her serum urea nitrogen was 75.9 mg/dl and creatinine was 5.4 mg/dl accompanied by oliguria. The myoglobin value was 27,000 ng/ml in serum and 88,000 ng/ml in urine. She was diagnosed as acute renal failure caused by rhabdomyolysis and hemodialysis therapy was started. She was released from hemodialysis on the 31th of January. The swelling of her right leg disappeared at the end of February. However, her right foot was affected paralysis of the fibular nerve. Electromyogram of her right anterotibial muscle and the test of conduction velocity of right tibial nerve revealed that the neurological disturbance of her right leg was caused by thrombosis of the deep vein. Generally speaking, the swelling of the extremities resulting from rhabdomyolysis caused by crush syndrome is due to a massive shift of body-fluid into the crushed muscles. We believe that when the extremities are compressed (and/or crushed) for a long time, venous thrombosis of the extremities occurs due to compression, there by causing swelling of the compressed extremities, as in this case.

Acute Kidney Injury↗

PubMed1998

Detection of DNA amplification in human renal cell carcinoma cell lines using restriction landmark genomic scanning.

Gene amplification, which has often been observed in various human cancers, appears to be associated with the development and progression of malignant phenotypes. However, in renal cell carcinoma (RCC), conventional analytic methods requiring specific primers and probes have revealed infrequent amplification of known oncogenes. We attempted to determine if gene amplification was truly uncommon in RCC. The genomic DNAs extracted from 5 human RCC cell lines were examined by restriction landmark genomic scanning (RLGS), a two-dimensional gel analysis which allows evaluation of approximately 2,000 radiolabelled DNA fragments. By this method, we detected 24 distinct spots commonly amplified in at least 2 RCC cell lines compared to normal kidneys. Comparing the present results with chromosomal assigned-RLGS, approximately one half of these DNA fragments proved to be located on chromosome 2, 5 or 7. Our data suggest that amplification of unknown genes is likely to occur in RCC cell lines.

Carcinoma, Renal Cell↗

PubMed1998

Autologous blood transfusion and hypotensive anesthesia for rotational acetabular osteotomy.

We employed hypotensive anesthesia with prostaglandin E1 (PGE1) and transfused 400 ml of autologous whole blood on the first postoperative day to avoid homologous blood transfusion in 137 patients undergoing rotational acetabular osteotomy (RAO) and RAO combined with intertrochanteric valgus osteotomy. Four hundred ml of whole blood were donated 1 week before the surgery, and transfused on the first postoperative day after hematological examination. We employed induced hypotension with PGE1 under general anesthesia with enflurane or isoflurane. Intraoperative systolic blood pressure was maintained at approximately 70-80 mmHg. The mean operation time was 139 +/- 32 minutes. The mean intraoperative and the postoperative estimated blood loss was 286 +/- 152 g and 259 +/- 122 g, respectively. The mean hemoglobin content was 13.3 +/- 1.1 g/dl before the operation. It showed the lowest value of 9.4 +/- 1.0 g/dl on the first postoperative day but it returned to 10.4 +/- 1.0 g/dl in the second postoperative week. There was no use of homologous blood transfusion. We consider that the concomitant use of autologous blood donation and hypotensive anesthesia is an extremely useful method to avoid homologous blood transfusion.

Acetabulum↗

PubMed1997

Retinoblastoma protein expression and MIB-1 correlate with survival of patients with malignant astrocytoma.

BACKGROUND: Regulatory coupling of cell proliferation and apoptosis is suggested by recent findings with regard to certain tumors, such as the finding of tumor resistance to anticancer therapy caused by inhibition of apoptosis. The processes leading to apoptosis appear to be regulated by a variety of oncogenes and tumor suppressor genes. In the current study, the relation between apoptosis and expression of retinoblastoma protein (pRB) was assessed in 50 primary anaplastic astrocytomas (AAs) and 46 recurrent tumors in the same patients as the primary tumors after macroscopic total surgical resection and chemoradiotherapy. METHODS: Apoptotic cells were identified by the in situ 3'-end labeling technique. Proliferative potential, pRB expression, and p53 expression were evaluated immunohistochemically using anti-Ki-67 (MIB-1), anti-pRB, and anti-p53 antibodies, respectively. The prognostic value of these biologic markers in AA patients undergoing treatment was also evaluated. RESULTS: The mean apoptotic index (AI) was 0.91 +/- 0.70% for specimens obtained at the initial surgery and 2.32 +/- 1.71% for those obtained at recurrence. There was no apparent correlation between the AI and the MIB-1 staining index (MI) in primary AAs, whereas significantly higher AI and MI were observed in recurrent pRB negative cases than in their pRB positive counterparts. The survival of patients with AAs showing a high MI and negative pRB immunostaining was significantly shorter than in the other cases. Neither the size of the apoptotic fraction nor the p53 expression in primary tumor correlated with the overall survival. CONCLUSIONS: The clinical outcome of patients with AA may be associated with aberrant pRB function and increased proliferative activity rather than an inability of tumor cells to undergo apoptosis.

Adult↗

PubMed1997

Comparison of ras activation in prostate carcinoma in Japanese and American men.

BACKGROUND: Comparative studies of point mutations in K-, N-, and H-ras oncogenes were performed on prostate carcinoma from Japanese and American patients to clarify the racial difference. METHODS: We probed for mutations in 70 Japanese and 31 American specimens using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis and immunohistochemistry for ras p21. RESULTS: Within the 70 Japanese specimens, eight mutations in codon 12 of K-ras (five GGT-->GTT transversions and three CGT-->GAT transitions) and one mutation in codon 12 of the N-ras gene (a GGT-->GTT transversion) were confirmed, whereas the American samples yielded only one definable mutation, a GGT-->GAT transition, in codon 12 of K-ras. CONCLUSIONS: The frequency of ras gene mutations in clinical carcinoma in Japanese men was higher than that in American men. It is suggested that there may be fundamental differences in the etiology of prostate carcinoma in Japan and the United States, perhaps based on genetics and/or environmental factors.

Adenocarcinoma↗

PubMed1997

Genomic alterations of human gliomas detected by restriction landmark genomic scanning.

Alterations of genomic DNA in eight primary astrocytic tumors and two glioma cell lines were examined using a recently developed two-dimensional gel electrophoresis method called restriction landmark genomic scanning (RLGS). RLGS allows us to detect amplifications, deletions, and methylation in genomic DNA in one procedure without requiring any polymorphic markers. Approximately 2000 spots (landmark sites) in tumor specimens were compared with those in normal brain tissue. The 10 spots with intensified signal were reproducibly detected in at least 50% of primary tumors, implying amplification of corresponding DNA sequences. Conversely, 12 spots with reduced signal were observed in more than 50% of all tumors, suggesting inactivation by allelic loss, homozygous deletion, or CpG island methylation. These results suggest that common genetic alterations are closely correlated with the genesis or progression of human gliomas.

Brain Neoplasms↗

PubMed1997

Analyses of human gliomas by restriction landmark genomic scanning.

The 16 primary gliomas were examined for changes in genomic DNA using a recently developed 2-dimensional gel electrophoresis method called restriction landmark genomic scanning (RLGS). This approach allows detection of DNA amplification, deletion, methylation and potentially other genetic rearrangements represented as decreases and increases in spot/fragment intensity on an autoradiogram. Approximately 2,000 landmark sites in tumor DNA were compared with those of DNA isolated from normal brain tissues. Seven spots showing intensified signal were consistently detected in at least 50% of tumors, implying activation of corresponding DNA sequences, and 8 additional spots having reduced signal were observed, again in more than 50% of all tumors, suggesting inactivation by the loss of 1 allele or homozygous deletion. Decreased signal may also infer relative CpG island methylation state. Of those spots consistently identified in tumors, 2 amplified and 4 reduced spots were found to be characteristic of low- and high-grade tumors, while the remaining 5 amplified and 4 reduced spots were associated with high-grade gliomas only, suggesting a link of specific mutations to degree of malignancy. A separate subset of glioblastomas evaluated, however, showed no alterations in these 'hot spots' which were detected in even low grade astrocytomas. The results demonstrate the genetic heterogeneity of glioblastoma and implicate the progression of neoplasia via differing genetic pathways.

Astrocytoma↗

PubMed1997

Genomic alterations in human glioma cell lines detected by restriction landmark genomic scanning.

Restriction landmark genomic scanning (RLGS) is a 2-dimensional gel analysis capable of detecting amplifications, deletions and rearrangements in genomic DNA. Using RLGS, we examined genomic DNA from each of 6 human-derived malignant glioma cell lines and from normal brain tissue samples. RLGS allows us to screen genomic DNAs as approximately 2,000 landmark sites in one procedure without any polymorphic markers. The resulting 2,000 spots in tumor samples were compared with those in normal brain. Six spots common to 5 of the 6 cell lines showed intensified signal, suggesting amplification of a tumor-specific DNA fragment. In addition, 25 spots common to 5 of the 6 lines showed a decrease in signal intensity, conversely indicating allelic loss of homozygous deletion. These results imply the existence of consistent genetic alterations in human glioma.

Brain Neoplasms↗

PubMed1997

Genetic alterations in N-bis(2-hydroxypropyl)nitrosamine-induced rat transplantable thyroid carcinoma lines: analysis of the TSH-R, G(alpha)s, ras and p53 genes.

It has recently been shown that point mutations of the TSH-R or G(alpha)s genes are associated with autonomous hyperfunctioning thyroid adenomas and differentiated carcinomas. We therefore screened for mutations in the TSH-R, G(alpha)s, ras and p53 genes in nine rat transplantable thyroid carcinoma lines derived from tumors induced by DHPN as a chemical carcinogen. Mutations were identified using single-strand conformation polymorphism and DNA sequencing analysis. Point mutations in G(alpha)s codon 201 (CGC-->CAC) were detected in three lines (33%), resulting in a heterozygous alteration (Arg-->His) in the expressed G(alpha)s protein. The mean intracellular cAMP level (2.30 +/- 0.27 nmol/mg) of the three mutated cell lines was significantly increased as compared with that of the lines (1.54 +/- 0.32 nmol/mg) without the G(alpha)s mutation (P < 0.01, by paired t-test). Also, these three cell lines had an activating mutation in Ki-ras codon 12 (GGT-->GAT). One TSH-R gene mutation was found with a base substitution in codon 636 (TGC-->TGT) but no amino acid change. No p53 gene (exons 5-8) mutations were detected in any of the cell lines analyzed. The results suggest that mutational activation of the G(alpha)s gene may play a tumorigenic role through constitutive activation of the cAMP pathway and that G-->A point mutations in the G(alpha)s and ras genes in thyroid carcinomas directly reflect interaction of the chemical carcinogen with guanine residues in DNA.

Animals↗

PubMed1997

Genetic changes in prostate cancer.

Recent advances in molecular biology have allowed us to understand that it is the accumulation of genetic alterations which leads to each step of tumorigenesis. What the specific alterations may be, however, often varies with each neoplasm. Prostate cancer is somewhat unique in its presentation to the pathologist of a bewildering array of histologies difficult to assign to diagnostic categories and contributing to misinterpretations of underlying molecular events. As with any malignancy, it is of utmost importance to thoroughly analyze and record the genetic aberrations found in prostate cancer with the objective of correlation to the pathology and natural history of the disease. Multiple oncogenes and tumor suppressor genes have been investigated in both clinical and latent cancer using conventional mutational analyses. To probe deeper into these genes and to uncover novel molecular events, genomic tumor DNA were examined using restriction landmark genomic scanning (RLGS), a method which allows the identification and comparison of specific genetic alterations within large segments and multiple samples of DNA at a time. This article reviews what has been identified based on numerous molecular studies, focusing on the genetic alterations peculiar to human prostate cancer.

Chromosome Deletion↗

PubMed1997

A case of Kawasaki disease with coronary artery aneurysms documenting Yersinia pseudotuberculosis infection.

A case of a 5-year-old boy who fulfilled all the criteria for Kawasaki disease (KD) was described. He had associated bilateral coronary artery aneurysms. Our study revealed the isolation of Yersinia pseudotuberculosis in stool cultures, and the elevation and seroconversion of the agglutination antibody titres, and hence he was diagnosed as Y. pseudotuberculosis infection-positive. We also demonstrated the positive mitogenic activity of the culture supernatant of the isolated bacterium from the patient and detected Y. pseudotuberculosis-derived mitogen by PCR. This case therefore suggests that Y. pseudotuberculosis might be closely related to the cause of KD.

Angiography↗

PubMed1997

Characterization of the molecules involved in the hematopoietic microenvironment provided by mouse stromal cell line MC3T3-G2/PA6 using a unique reporter system that analyzes the direct cell-to-cell interaction.

As an approach to characterizing the molecules involved in the hematopoietic microenvironment provided by a murine clonal preadipose cell line MC3T3-G2/PA6 (PA6), we developed a unique system to detect the early phase of signal transduction caused by the direct cell-to-cell interaction using the reporter plasmid pfosluc2 with the c-fos enhancer/promoter linked with the Photinus pyralis luciferase gene. The plasmid pfosluc2 was genetically introduced into a mouse myeloid leukemia cell line NFS-60 which showed a growth dependency on contact with PA6 cells, and the mechanism by which stromal PA6 cells promote the proliferation of NFS-60 cells through the direct cell-to-cell interaction was analyzed. The direct cell-to-cell interaction with PA6 cells was found to cause a significant c-fos induction to NFS-60 cells within 1 h. Approximately 10(5) cDNA clones prepared from PA6 cells were screened for their activity to promote the c-fos expression in NFS-60 cells through the direct cell-to-cell interaction, and 13 positive clones were obtained. Of these positive clones, five clones encoded the stem cell factor, and the others encoded the hepatocyte growth factor (HGF). The c-fos induction caused by the contact with PA6 cells in NFS-60 was completely inhibited by addition of both antagonistic anti-c-kit and anti-HGF antibodies. These results represent direct evidence for the action of HGF on the proliferation of hematopoietic cells through direct cell-to-cell interaction with stromal cells. Thus, our developed reporter system can be useful in investigating the direct cell-to-cell interaction between stromal and hematopoietic cells.

Animals↗

PubMed1997

Studies on anti-inflammatory agents. V. Synthesis and pharmacological properties of 3-(difluoromethyl)-1-(4-methoxyphenyl)-5- [4-(methylsulfinyl)phenyl]pyrazole and related compounds.

A series of novel 1,5-diphenylpyrazole derivatives bearing hydrophilic substituents was prepared. The anti-inflammatory and analgesic activities of these compounds were evaluated by using the adjuvant arthritis and Randall-Selitto assays in rats, and the structure-activity relationships were studied. The optimal compound was 3-(difluoromethyl)-1-(4-methoxyphenyl)-5-[4-(methylsulfinyl)phenyl]pyraz ole (10) with oral ED50 values of 0.31 and 2.6 mg/kg on adjuvant-induced arthritis and carrageenin-induced foot edema, respectively. Compound 10 showed analgesic activities not only toward inflamed paw but also toward normal paw (ED30 = 0.55 and 1.8 mg/kg, respectively) in the Randall-Selitto assay, and moreover, 10 was effective in the tail-pinch assay (ED50 = 21 mg/kg) similarly to morphine. The asymmetric synthesis and pharmacological properties of the enantiomers of 10 are also reported.

Animals↗

PubMed1997

Studies on anti-inflammatory agents. IV. Synthesis and pharmacological properties of 1,5-diarylpyrazoles and related derivatives.

A series of novel 1,5-diarylpyrazole derivatives was synthesized and tested for anti-inflammatory and analgesic activities to develop anti-inflammatory agents with fewer side effects than existing nonsteroidal anti-inflammatory drugs. The structure-activity relationships in this series were extensively studied. Electron-withdrawing substituents such as CN and CF3 were optimal at the 3-position of the pyrazole ring. Replacement of these substituents with bulky ones gave less active compounds. The 4-(methylsulfonyl)phenyl group seemed to be the optimal group at the 5-position of the pyrazole ring. The most potent compound was 1-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]-pyrazole-3-carbonitrile (19a), with oral ED50 value of 0.030 and 0.47 mg/kg on adjuvant-induced arthritis and collagen-induced arthritis, respectively, and an ED30 value of 7.4 mg/kg in the yeast-induced hyperalgesia (Randall-Selitto) assay. Compound 19a also showed potent inducible cyclooxygenase (COX-2)-inhibitory activity (IC50 = 0.24 microM) with no COX-1 inhibition even at 100 microM.

Anilides↗

PubMed1997

Different patterns of DNA alterations detected by restriction landmark genomic scanning in heterogeneous prostate carcinomas.

Alterations in the genomic DNAs of six heterogeneous prostate carcinomas, as well as that of individual and histologically distinct foci within the tumors, were examined using restriction landmark genomic scanning, a method employing two-dimensional gel analysis of a large number of DNA fragments generated by digestion with highly specific endonucleases. Upon autoradiographic imaging, these fragments appear as spots of varying intensity and location specific for each sample. In our study, comparison of cancer DNAs against normal prostate DNA controls yielded alterations in at least 35 spots. Despite differences in the histological grading of tumors, 3 spots common to all tumor samples showed consistent amplification of intensity and 8 other common spots demonstrated consistent reduction of intensity when compared with control. In addition, spot alterations occurred between histologically identical foci isolated from within single tumors. We suggest that these spot changes detected in DNA profiles generated by restriction landmark genomic scanning reflect aberrations in as yet unidentified oncogenes and tumor suppressor genes and indicate that prostate cancer is not only histologically heterogeneous and multifocal but also genetically multicentric.

Aged↗

PubMed1997

Genetic variations in human benign prostatic hyperplasia detected by restriction landmark genomic scanning.

Alterations in the genomic DNAs of benign prostatic hyperplasias were examined using restriction landmark genomic scanning. This methodology allows the visual comparison of a large number of loci as spots within a gel that has been electrophoresed in 2 dimensions. Within each of the 16 hyperplasias examined, we found 2 to 10 spots with altered intensities, as compared to equivalent spots from normal prostate, but could detect few common spot profiles for any hyperplasia samples. These data conflict with data from previous studies and suggest that there may be no consistent genetic changes in the pathogenesis of some forms of human benign prostatic hyperplasia.

Aged↗

PubMed1997

[A case report of spontaneous hemopneumothorax].

A 55-year-old man was admitted to the private hospital with chief complaints of left chest pain and increasing dyspnea. Chest X ray on admission revealed a collapsed lung and an air fluid line in the left hemothorax. A chest tube drainage was carried out, but hemorrhagic pleural fluid of 1,000 ml was drainaged. So was the patient transferred to our hospital for surgical treatment. Following blood transfusion because of hemorrhagic pleural fluid of 1,500 ml, urgent surgery was carried out. At the operation under thoracoscope, the bleeding from the apex of the thoracic cavity was noted. The bleeding was controlled with endoscopic surgical clip. As it was, however, very difficult to remove massive blood clots under endoscopic surgery, a small thoracotomy was carried out in addition to the thoracoscopic surgery, then the bulla in the apex of left lung was resected and also removed a large amount of blood clots.

Hemopneumothorax↗