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T Kishida

Publications and source records attributed to T Kishida.

At least 19 recordsLinked to original sources
PubMed1995

Suppression of growth of renal carcinoma cells by the von Hippel-Lindau tumor suppressor gene.

Clear cell renal carcinomas are most frequently characterized by loss of function of both copies of the von Hippel-Lindau (VHL) disease gene, suggesting that the VHL gene product plays an important role in regulating renal cell proliferation. To directly assess the function of the VHL gene product, we transfected the wild-type VHL gene into two renal carcinoma cell lines that lacked normal expression of the gene. Expression of the wild-type VHL gene led to a dramatic suppression of growth in two renal carcinoma cell lines, A498 and UMRC6 in vitro, as measured by colony formation and direct cell counting. Transfection of a naturally occurring mutant VHL gene (nucleotide 713 G to A, Arg to Gln) did not lead to growth suppression of these renal carcinoma cells, nor did transfection of the wild-type VHL gene into two non-renal tumor cell lines that expressed the endogenous wild-type VHL gene. Expression constructs, which included the first ATG at nucleotide 214, were sufficient to produce the strongest growth suppression. These experiments provide direct evidence that the VHL gene product functions to suppress the growth of renal carcinoma cells and also provide a model for mapping the domains of the VHL protein important in suppressing tumor growth.

Base Sequence↗
PubMed1995

Cellular proteins that bind the von Hippel-Lindau disease gene product: mapping of binding domains and the effect of missense mutations.

The von Hippel-Lindau disease (VHL) gene is a novel tumor suppressor gene that plays a role in the pathogenesis of renal cell carcinomas and hemangioblastomas of the central nervous system. To begin an evaluation of the biological functions of the VHL gene product (pVHL), we prepared bacterial fusion protein between glutathione S-transferase and wild-type or mutant pVHLs. The fusion proteins were used to identify cellular proteins that bind to pVHL in vitro. Monkey kidney cells transfected with wild-type or mutant VHL cDNAs were used to identify cellular proteins that bind to pVHL in vivo. Wild-type pVHL consistently bound two cellular proteins with apparent molecular masses of 10 and 14 kilodaltons that were designated p10 and p14, respectively. Mapping studies with a panel of VHL deletion mutant proteins demonstrated that p10 and p14 bound to a 32-amino acid peptide located in the carboxy terminal portion of pVHL. Missense mutation located within this 32-amino acid peptide abrogated the ability of the VHL protein to bind p10 and p14. Of 67 VHL families with identified germline mutations, 42 families had mutations predicted to affect the p10/p14-binding region. Maintenance of the integrity of the p10/p14-binding region appears to be essential for cellular growth regulation by pVHL.

Animals↗
PubMed1995

Germline mutations in the von Hippel-Lindau disease tumor suppressor gene: correlations with phenotype.

von Hippel-Lindau disease (VHL) is an inherited neoplastic disease characterized by a predisposition to develop retinal angiomas, central nervous system hemangioblastomas, renal cell carcinomas, pancreatic cysts, and pheochromocytomas. The VHL gene was recently isolated by positional cloning. The cDNA encodes 852 nucleotides in 3 exons. The VHL gene is unrelated to any known gene families. We identified germline mutations in 85/114 (75%) of VHL families. Clinical heterogeneity is a well-known feature of VHL. VHL families were classified into 2 types based on the presence or absence of pheochromocytoma. The types of mutations responsible for VHL without pheochromocytoma (VHL type 1) differed from those responsible for VHL with pheochromocytoma (VHL type 2). Fifty-six % of the mutations responsible for VHL type 1 were microdeletions/insertions, nonsense mutations, or deletions; 96% of the mutations responsible for VHL type 2 were missense mutations. Specific mutations in codon 238 accounted for 43% of the mutations responsible for VHL type 2. The mutations identified in these families will be useful in presymptomatic diagnosis. The identification of mutations associated with phenotypes contributes to the understanding of fundamental genetic mechanisms of VHL disease.

Adrenal Gland Neoplasms↗
PubMed1995

Von Hippel-Lindau (VHL) disease with pheochromocytoma in the Black Forest region of Germany: evidence for a founder effect.

We identified a germline missense mutation at nucleotide 505 (T to C) of the VHL tumor suppressor gene in 14, apparently unrelated, VHL type 2A families from the Black Forest region of Germany. This mutation was previously identified in two VHL 2A families living in Pennsylvania (USA). All affected individuals in the 16 families shared the same VHL haplotype indicating a founder effect. This missense mutation at codon 169 (Tyr to His) would probably cause an alteration in the structure of the putative VHL protein. The association of this distinct mutation with the pheochromocytoma phenotype in VHL may help to elucidate the genetic mechanism of carcinogenesis in this multi tumor cancer syndrome.

Adrenal Gland Neoplasms↗
PubMed1995

Diagnosis of preterm premature rupture of the membranes using a newly developed AFP monoclonal antibody test kit.

OBJECTIVE: We have developed a new anti-AFP monoclonal antibody kit which is easier to use than other examination methods for detecting AFP in the leaked amniotic fluid. In this study, we investigate the clinical value of this test in the diagnosis of preterm premature rupture of the membranes (PROM). METHODS: We employed 103 patients of less than 37 weeks of gestational age for this preliminary study. We compared the clinical usefulness of this new AFP test with that of the nitrazine test and measured the concentration of AFP in vaginal fluid or cervical secretion by EIA. RESULTS: The nitrazine test showed a correct diagnostic rate of 62.1%, in contrast the AFP test kit had a 98.0% rate (P < 0.001). The reaction time using the kit is 3 min. CONCLUSION: The AFP test is a simple and non-invasive test which can be easily carried out repeatedly as a bedside examination. This study has confirmed the high clinical efficacy of the newly developed AFP test kit as a method of PROM diagnosis.

Amniotic Fluid↗
PubMed1995

[Prognostic factors for metastatic renal cell cancer].

We examined various prognostic factors of metastatic renal cell carcinoma. Patients who had metastasis at nephrectomy (A group, 38 cases) and those who had metastasis as recurrent tumors after nephrectomy (B group, 38 cases) entered in this study. Five-year survival rate of total cases after confirmation of metastatic foci was 15% and there was no statistical significant difference between A and B groups. Several pathological factors were related to poorer prognosis and included large diameters of original tumors, positive lymph nodes, higher grade tumors and non-clear cell type tumors. Patients who have a solitary lung metastasis showed better prognosis compared to those with multiple lung metastases or metastases of other organs. Two factors related to treatment were shown to contribute to better prognosis. These were the response to interferon alfa (IFN alpha) and the possibility of total resection of visible metastatic tumors. Patients who belong to A group were shown to achieve markedly better therapeutic benefit from IFN alpha or IFN alpha plus anticancer drugs. Five-year survival rate for the responders was 40%, as compared to less than 5% for the non-responders. Ten-year survival rate for patients with metastasis who had undergone complete resection of visible tumor was 50%, and the for patients belonging to B group Showed 80%. We concluded that these prognostic factors should be considered to decide how to select patients with metastatic renal cell cancer.

Adult↗
PubMed1995

Preparation of multi-locus DNA probe cocktail by liquid-phase reassociation.

We developed a simple, rapid method for the preparation of a DNA-fingerprinting probe cocktail, and tested its usefulness in paternity testing. Exploiting the property of tandemly repetitive DNA segments to be rapidly renatured after heat denaturation, we enriched restriction fragments of a child's genomic DNA for minisatellites by liquid phase reassociation followed by capture with immobilized streptavidin. We amplified and simultaneously labeled the reassociation product by anchored PCR using a digoxigenin-labeling mixture. Using this probe cocktail, we were able to detect fingerprints of paternity case trios, and the results were corroborated by DNA fingerprinting with a commercially available probe as well as by conventional phenotyping. Our method enables one to prepare a fresh cocktail of probes from the DNA sample under study during the overnight electrophoresis and Southern transfer steps in DNA fingerprinting, and eliminates the need of having an expensive probe of limited shelf life. If one has a practical outlook on DNA fingerprinting and regard it as a preliminary test, one does not have to use a cloned DNA probe. The present study demonstrates that a multi-locus probe cocktail serves such a practical purpose.

Base Sequence↗
PubMed1995

Retinoblastoma gene mutation in primary human renal cell carcinoma.

We searched for possible mutations in the E2F-binding region of retinoblastoma gene in primary human renal cell carcinomas, using polymerase chain reaction and single-strand conformational polymorphism analysis of RNA. Retinoblastoma gene mutation was detected in 1 of 21 cases (5%). DNA sequencing of the polymerase chain reaction product verified that this case had a 6-base deletion at the beginning of exon 8. Our findings suggest that mutation of the retinoblastoma gene is involved in only a subgroup of sporadic human renal cell carcinomas.

Base Sequence↗
PubMed1995

Diagnosis of premature rupture of membranes with an improved alpha-fetoprotein monoclonal antibody kit.

We developed a new kit for detecting alpha-fetoprotein (AFP) in leaked amniotic fluid (Eur J Obstet Gynecol Reprod Biol 1995;58:67-72). Later, we developed an improved AFP kit utilizing the same AFP monoclonal antibody. We compared this improved AFP test with the nitrazine test for 137 patients. The nitrazine test correctly diagnosed 62.1% of the cases, but the improved AFP kit diagnosed 98.0% for < 37 weeks of gestation (P < 0.001). The nitrazine test showed a specificity of 58.3%, whereas the AFP kit showed a 100% rate for detecting > or = 37 weeks of gestation (P < 0.01). The reaction time with the AFP kit is 90 s. This study has confirmed a high clinical efficacy of the improved AFP test kit as a method of diagnosis of premature rupture of fetal membranes.

Amniotic Fluid↗
PubMed1994

[Definite diagnosis of premature rupture of the membranes (PROM) using intra-amniotic dye injection method and development of a new kit (AFP-test kit) for PROM diagnosis].

In the diagnosis of premature rupture of the membranes (PROM) in the mid-trimester of pregnancy, it is often difficult to obtain reliable information from only an analysis of the amniotic fluid, especially in equivocal cases. In cases where we have not been able to obtain a definite diagnosis of PROM by conventional methods including amnioscopy, we have employed the intra-amniotic dye injection method (PSP method) by abdominal amniocentesis. We have established the method to be good for discrimination of high-leak and low-rupture PROM as well as the rupture of the pseudo-amniotic cavity, and have preliminarily reported on the clinical usefulness and safety of the PSP method since 1981. In this study, through examination of 64 equivocal PROM cases, we have investigated the clinical efficacy of the PSP method for cases in their 14th to 33rd week of gestation. The conventional methods showed a correct diagnostic rate of 63.9%-70.5%, in contrast the PSP method had a 100% rate. We have reconfirmed the clinical efficacy of the PSP method. The PSP method has been proven to be a reliable PROM diagnostic method, but employed for a selected doubtful cases. A non-invasive, reliable and rapid method which can be employed in a repeated manner as a bed-side examination, has been needed for many years. Amniotic fluid contains a high concentration of alpha-fetoprotein(AFP), especially in mid-trimester, and undetectable levels are determined in urine, vaginal fluid and seminal fluid. We have developed a new anti-AFP monoclonal antibody kit for PROM diagnosis. In the present study, we investigated the fundamental ability of AFP-test kit and the clinical efficacy of this kit for 71 cases in their 11th to 40th week of gestation with PROM or suspected PROM. The AFP-test kit method showed a correct diagnostic rate of 100%. The reaction time of this kit is approximately 3 minutes. It is a simple and non-invasive test which can be easily carried out repeatedly as a bed-side examination. This study has confirmed the high efficacy of the AFP-test kit as a method of PROM diagnosis.

Amniocentesis↗
PubMed1994

Universal probe system for DNA fingerprinting.

A universal probe system is a combination of an unlabeled primary probe that has been inserted in a cloning vector and a labeled secondary probe that is specific to the vector. This system is time- and labor-saving in that one does not need to label probes every time one uses them, so long as they are inserted in the same vector. As a first step in the preparation of a universal probe system for DNA fingerprinting, we isolated multi-locus probes from a subgenomic library that had been constructed by insertion into the phagemid pUC118 of 1-2 kb fragments of DNA from human myeloma cells. Next, we isolated single-stranded DNAs of recombinant phagemids, and hybridized them with Southern blots of DNAs from mother-child-father trios or unrelated individuals. As with double-stranded DNA probes, we were able to detect DNA fingerprints, with a commercially available, alkaline phosphatase-labeled secondary probe or a digoxigenin-labeled universal sequencing primer.

DNA Fingerprinting↗
PubMed1992

A preliminary study on the effect of alpha-interferon treatment on the joint inflammation and serum calcium in rheumatoid arthritis.

A 12-week, double-blind controlled study comparing low dose alpha-interferon (5 x 10(5) iu, twice a week) and placebo was carried out to determine whether this drug has an immunomodulatory effect in rheumatoid arthritis. There were statistically significant improvements in the patients' joint score, C-reactive protein value and platelet count, without any side effects. Interferon-alpha caused an increase of serum calcium and a decrease of alkaline phosphatase. Thus, alpha-interferon requires further investigation as a possible additional useful treatment for rheumatoid arthritis.

Adult↗
PubMed1992

An endoscopic study of antibiotic-associated hemorrhagic colitis.

Colonoscopy was performed on acute stage within 72 hours from onset in 48 patients with antibiotic-associated hemorrhagic colitis (AAHC). From our observations we have developed a new endoscopic classification of AAHC and investigated the incidence of each finding in further detail. We have classified in principle the endoscopic findings of the 48 subjects into two groups: major and minor ones respectively. Category of major findings were: 1) diffuse mucosal hemorrhage (100%); 2) spotty mucosal hemorrhage (100%), and 3) linear mucosal hemorrhage (22.9%), while minor findings were: 1) irregular ulcers in 10.4%; 2) aphthoid ulcers in 6.3%, and 3) linear erosions or ulcers in 4%. Minor findings were ulcers or erosions present over the hemorrhagic mucosa associated with the moderate degree of inflammation. A histopathologic study of colon biopsy specimens from 24 patients with AAHC showed hemorrhage and inflammatory cell infiltration in the lamina propria mucosae varying from mild to moderate in extent. It was concluded that AAHC was a colonic mucosal hemorrhagic disease caused by the destruction of mucosal vessels from unknown causes and in this disease mild to moderate inflammation was partially followed by ulceration over the edematous and hemorrhagic mucosa of the colon.

Adolescent↗
PubMed1992

Pulmonary infiltration and eosinophilia associated with sulfasalazine therapy for ulcerative colitis: a case report and review of literature.

We report a 52-yr-old man with ulcerative colitis who developed sulfasalazine-induced pulmonary infiltration with eosinophilia (PIE syndrome), which resolved completely after withdrawal of this drug. Desensitization to sulfasalazine was successful, and allowed the patient to receive this drug without recurrence of the pulmonary toxicity. This is the first case of the sulfasalazine-induced PIE syndrome in Japan; a review of the world literature found no previous cases of successful desensitization following sulfasalazine-induced PIE syndrome.

Colitis, Ulcerative↗
PubMed1992

Efficient methods for production of monoclonal antibodies for forensic applications.

To simplify the labor-intensive process of making hybridomas, we fused popliteal lymph node cells, used an autoclavable serum-free culture medium throughout, and cloned hybridomas with human blood cell feeders. To make the best possible use of the serum-free medium, we adapted mouse myeloma cells to it. Using as little as 0.2 ml of human blood per culture plate, we successfully cloned hybridomas and established a hybrid cell line producing anti-peroxidase antibody. Our protocol affords a considerable saving on time, labor, and cost, and hopefully will encourage the forensic scientist to undertake the production of monoclonal antibodies.

Animals↗