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N Tsuyama

Publications and source records attributed to N Tsuyama.

At least 19 recordsLinked to original sources
PubMed1998

A hVti1 homologue: its expression depends on population doubling levels in both normal and SV40-transformed human fibroblasts.

A cDNA clone was isolated by differential colony hybridization from a cDNA library prepared from life-extended SV40-transformed human fibroblasts. The clone, tentatively named N-10, was 1272 bp in length coding for 232 amino acids. Northern analysis revealed that the expression level of N-10 was increased in normal senescent and life-extended SV40-transformed fibroblasts than in their young counterparts but was not enhanced by growth arrest. The protein fused to GFP (green fluorescent protein) localized in cytoplasmic granule. Enforced expression of N-10 resulted in premature senescence in young fibroblasts. The deduced amino acid sequence of N-10 was identical to the recently reported hVti1 gene except in one amino acid: Asp24(GAC) was ours and Asn24 (AAC) was reported. Additional base differences were found, so we referred to our sequence as the hVti1 homologue. As hVti1 protein was suggested to be involved in the vesicle transport process, the homologue may be concerned with increased secretion of extracellular matrix and various cytokines associated with cellular senescence.

Carrier Proteins↗
PubMed1996

Gain-of-function p53 mutations enhance alteration of the T-cell receptor following X-irradiation, independently of the cell cycle and cell survival.

Missense mutations are by the far the most common types of mutations found in p53 of human tumors, suggesting that mutant p53 proteins function either by abrogating wild-type function or by gaining new oncogenic functions. To distinguish between the dominant-negative effect and gain of new function of p53 missense mutants, we measured the ability of transfected missense mutant p53s in p53-null Jurkat cells to alter T-cell receptor (TCR) surface expression. The TCR is a key signal transduction moiety common to T lymphocytes and is one of the major sites for aberrations in T-cell leukemias/lymphomas. Three p53 mutants (248trp, 249ser, and 273his) enhanced the frequency of TCR mutants after graded doses of X-radiation compared to null p53 parent- and wild-type p53-possessing normal lymphocytes; the parent Jurkat and normal lymphocyte showed no difference. These enhancements were not the results of a change in radiosensitivity or in G1 checkpoint arrest characteristics. Therefore, the creation of this mutator phenotype by missense-type p53 mutations implies that a more direct mechanism, apart from changes of cell cycle kinetics or cell death, may be responsible for the selection of certain p53 point mutations, which eventually result in the tumorigenesis of the cell.

CD3 Complex↗
PubMed1995

Increase in expression levels of interferon-inducible genes in senescent human diploid fibroblasts and in SV40-transformed human fibroblasts with extended lifespan.

The normal human fibroblast line, TIG-3 which senesces at around 80 population doubling levels (PDLs), expressed interferon (IFN)-inducible genes such as 6-16, 2', 5'-oligoadenylate synthetase (2,5-A) and HLA B7 near the end of the proliferative lifespan. Other normal fibroblast line such as MRC-5 also expressed IFN-inducible genes when senesced. Clones transformed with SV40 T-antigen, which extended their proliferative lifespan by about 20-30 PDLs, also expressed IFN-inducible genes during their extended life. Anti-IFN-beta antibodies added in culture medium repressed the expression of IFN-inducible gene in both normal senescent and life-extended SV40-transformed cells. IFN-beta repressed DNA synthesis in normal TIG-3 and induced IFN-inducible genes in both normal and SV40-transformed TIG-3. Conditioned medium recovered from life-extended SV40-transformed cells contained IFN-beta, but not IFN-alpha, IFN-gamma or TNF-alpha and possessed an activity that inhibited DNA synthesis of young TIG-3. Addition of anti-IFN-beta antibodies into the medium enhanced the serum-induced DNA synthesis of near senescent (91% lifespan completed) TIG-3, while it neither induced DNA synthesis in fully senescent TIG-3 nor extended the proliferative lifespan of TIG-3. These results suggest that normal and SV40-transformed human fibroblasts increase expression of IFN-beta with increasing proliferative age especially near the end of their lifespan resulting in induction of IFN-inducible genes and possibly in growth repression.

Cell Line↗
PubMed1995

The usefulness of severe combined immunodeficiency (SCID) mice to study human carcinogenesis.

In the present study, we engrafted normal colonic epithelial and histologically diagnosed colonic adenomas from a familial adenomatous polyposis (FAP) patient into severe combined immunodeficient (SCID) mice and subsequently examined them histologically and molecular biologically. Successful engraftment and metastasis was observed. The facts that human normal colonic epithelium and adenomatous polyps can take in SCID mice indicates the possibility that this human SCID mouse system will be useful for investigating the dynamics of human carcinogenesis in various tissues.

Adenomatous Polyposis Coli↗
PubMed1994

Preparation of a subtractive cDNA library enriched in cDNAs which expressed at a high level in cultured senescent human fibroblasts.

Subtracted cDNA library was prepared by subtracting [cDNA from young growing SV40-transformed human fibroblasts] from [cDNA from growing SV40-transformed fibroblasts in extended lifespan]. Isolated cDNA clones which expressed at high level in life-extended transformed cells also expressed at high level in normal senescent fibroblasts but did at low level in growing and growth-arrested young cells. Neither fibronectin nor procollagen cDNA was isolated. This cDNA library is useful for isolation of senescent-specific cDNA species which express at high level in normal senescent cells but at low level in growing and growth-arrested young cells, avoiding growth-arrest-specific cDNAs.

Animals↗
PubMed1993

In vitro irradiation is able to cause RET oncogene rearrangement.

Elevated risk of thyroid cancers among the atomic bomb survivors as compared to the nonexposed population suggests that some genetic events related to thyroid cancer must be caused by ionizing radiation. Accordingly, inducibility of RET oncogene rearrangements, i.e., the generation of the RET-PTC oncogene, specific for thyroid cancer, was investigated among human undifferentiated thyroid carcinoma cells (8505C), which do not have RET oncogene rearrangement, after 0, 10, 50, and 100 Gy of in vitro X-irradiation by means of reverse transcription polymerase chain reaction. After testing 10(8) cells at each dose point, 3 independent samples obtained with 50 Gy of X-irradiation and 6 independent samples obtained with 100 Gy of X-irradiation showed a rearranged RET oncogene amplified band. No rearranged transcripts were obtained from cells irradiated with 0 or 10 Gy. All of the transcripts were sequenced and found to contain the D10S170 and RET sequence. Interestingly, two types of rearrangements were included in these transcripts: one is specific for thyroid cancer and the other, which contains a 150-base pair insert, is atypical, not usually seen in vivo. This insert was found to be the exon of D10S170. Furthermore, in fibrosarcoma cells (HT1080), X-irradiation also induced RET oncogene rearrangements, which included the same two types of rearrangements observed in the X-irradiated thyroid cells (8505C). These results are in favor of the hypothesis that some radiation-induced thyroid cancers, including those among atomic bomb survivors, might have developed when a growth advantage was obtained through a specific form of RET oncogene rearrangement induced by radiation exposure.

Aged↗
PubMed1993

DNA-DNA subtractive cDNA cloning using oligo(dT)30-Latex and PCR: identification of cellular genes which are overexpressed in senescent human diploid fibroblasts.

We previously reported an efficient method for subtractive cDNA cloning using oligo(dT)30-Latex and polymerase chain reaction (PCR) (E. Hara et al., Nucleic Acids Res. 19, 7097-7104, 1991). The subtraction was performed by hybridization between mRNA of cell type B and the cDNA made from mRNA of cell type A using an oligo(dT)30 primer covalently linked to Latex particles in an Eppendorf tube. The mRNA common to both types of cells could be removed by a brief centrifugation. In the present paper, the method was improved by using the sense strand DNA instead of mRNA for hybridization to cDNA covalently linked to the particles to minimize mRNA degradation and by optimizing the hybridization condition. The sense strand DNA was made from cDNA-oligo(dT)30-Latex by asymmetric PCR. Using the improved method, a subtractive cDNA library with longer cDNA inserts was successfully constructed with higher probability than the original method.

Animals↗
PubMed1993

Genetic alterations in thyroid tumor progression: association with p53 gene mutations.

To identify the genetic events that must be involved in thyroid tumor progression, we initially investigated p53 gene alterations in 10 papillary adenocarcinomas, 4 follicular adenocarcinomas, and 8 undifferentiated carcinomas. Base substitutional mutations in exons 5 to 8 and loss of heterozygosity (LOH) of the p53 gene were not detected in papillary or follicular adenocarcinomas. However, 7 of 8 undifferentiated carcinomas were carrying base substitutional mutations, and LOH was detected in 3 of 5 informative cases. Furthermore, to verify that the p53 gene alterations are truly involved in tumor progression, DNA from individual foci of the four undifferentiated carcinomas coexisting with a differentiated focus and from one follicular adenocarcinoma with an undifferentiated focus was analyzed by direct sequencing and polymerase-chain-reaction-restriction-fragment-length polymorphism (PCR-RFLP). Base substitutional mutations in the p53 gene from exons 5 to 8 were identified exclusively in the undifferentiated foci, but not in the differentiated foci. LOH was observed in 3 of 4 informative undifferentiated foci. In one of these positive cases, LOH was observed in both papillary adenocarcinoma and undifferentiated carcinoma. However, a p53 gene mutation at codon 248 was detected in the undifferentiated carcinoma but not in the papillary adenocarcinoma. The results imply that LOH occurs first in papillary adenocarcinoma followed by a p53 mutation during the transition from papillary adenocarcinoma to undifferentiated carcinoma. Maintenance of LOH during tumor progression excludes the possibility that these different histological foci are derived from different origins and represents molecular evidence that undifferentiated carcinoma is very likely derived from preexisting papillary adenocarcinoma. Furthermore, these results strongly suggest that the mutated p53 gene plays a crucial role in de-differentiation during the progression of thyroid tumors.

Adenocarcinoma↗
PubMed1992

Unique association of p53 mutations with undifferentiated but not with differentiated carcinomas of the thyroid gland.

Thyroid neoplasms show a wide variety of lesions varying from slowly growing differentiated adenocarcinomas to rapidly proliferating undifferentiated carcinomas. There has been some histopathological evidence that the undifferentiated thyroid carcinomas are derived from differentiated carcinomas. Moreover, it is suspected that some genetic events might be associated with such changes. In the present study, mutations in the p53 gene were investigated by direct sequencing analysis after polymerase chain reaction amplification of exons 5 to 8, using paraffin-embedded primary tumors and cultured cells. No mutations in exons 5 to 8 were detected in 10 differentiated papillary adenocarcinomas, whereas 6 of 7 undifferentiated carcinomas were found to carry base substitution mutations. Sequencing analysis confirmed mutations at codons 135 (TGC----TGT), 141 (CCC----CCT), 178 (CAC----GAC), 213 (CGA----TGA), 248 (CGG----CAG, CGG----TGG), and 273 (CGT----TGT). The spectrum of mutations (G:C to A:T transitions in 7 of 8) might be a specific feature of the spontaneous cancers. The results strongly suggest that, in human thyroid glands, p53 mutations play a crucial role in the progression of differentiated carcinomas to undifferentiated ones.

Adenocarcinoma, Papillary↗
PubMed1991

SV40 T-antigen is required for maintenance of immortal growth in SV40-transformed human fibroblasts.

Two lines of immortal human fibroblasts were isolated following transfection of TIG-3 cells with plasmid DNA, pMT-1ODtsA, that contained SV40 early gene with a deletion in replication origin and ts mutation in coding sequence for T-antigen. These cells continued proliferation at 34 degrees C, over 565 population doubling level (PDL) which is far over the limited division potential of untransformed normal TIG-3 of 70-80 PDL. When the culture temperature was shifted to 40 degrees C after 70 PDL, they ceased proliferation immediately. One of these immortal clones, SVts8, lost its ts phenotype after retransformation with wtT-antigen gene. These results indicated that the function of intact T-antigen is required for maintenance of immortal proliferation, at least in one of the SV40 transformed immortal clones.

Antigens, Polyomavirus Transforming↗
PubMed1990

[Spinal flexibility and activities of daily living for the thoracic and lumbar spinal cord injured--comparison of a non operated group with an internal spinal fused group].

There has been a great argument in selecting conservative treatment or surgical repair for the spinal cord injury as an early stage treatment. The purpose of medical treatment is to bring a patient back to the society as soon as possible by early intervention and rehabilitation, while preventing complications of the injury. We are faced with a difficulty in determining the superiority between conservative treatment and surgical repair, since no comparative statistical analysis has been available among various rehabilitation methods for the spinal cord injury. We treated 171 patients in the past with traumatic thoracic or lumbar spinal cord injury who were submitted to our hospital for the ADL training purposes. We selected and studied 34 complete paraplegic cases with no complications who started receiving ADL training within 2 months period after the injury from among these cases. (1) Non-ope group was superior in spinal flexibility to internally spinal fused group. (2) Non-ope group reached ADL independence 1.4 months earlier than I.S.F. in case of upper thoracic injury. ADL independence was reached almost at the same time in the lower thoracic injury cases by either treatment. Above all, it is concluded that conservative treatment is more helpful to establish rehabilitation in shorter length of time than surgical repair.

Activities of Daily Living↗
PubMed1989

Usefulness of myelography in brachial plexus injuries.

Ninety brachial plexus lesions have been examined by myelography and the results classified into six types. These were compared against the level of lesion found at exploration of the brachial plexus with electrophysiological investigations carried out during the operation. The results show that myelography can be a reliable and useful pre-exploratory measure to assess the level of the lesion of each injured root.

Adult↗
PubMed1989

Direct nerve crossing with the intercostal nerve to treat avulsion injuries of the brachial plexus.

One hundred seventy-nine patients with root avulsion brachial plexus injuries were treated with direct nerve crossing with the intercostal nerve and 159 cases were followed more than 1 1/2 years after the operation. When suture was done to the musculocutaneous nerve, 90% of 10 children who had operation within 7 months of injury and 81.8% of 110 adults, younger than 40 years with operation within 6 months of injury regained grade 3 or 4 elbow flexion power. This direct method seems to produce better results than those of nerve crossing, which uses intermediary nerve grafts.

Adolescent↗
PubMed1987

Expiratory activity in transferred intercostal nerves in brachial plexus injury patients.

Involuntary activity of transferred intercostal motor units was examined in patients with brachial plexus injury. Since the internal intercostal nerves were detached from the thorax to reinnervate the musculus biceps brachii, it was possible to record pure intercostal motor activity in humans. Respiratory activity was seen in the latter part of the expiratory phase, thus dividing the phase into two substages (E1 and E2) by the onset of the activity. CO2 rebreathing prolonged the duration of the intercostal motor activity and increased the tidal activity as determined from the integration curve. There was a close linear correlation between these two variables. These observations indicate that expiratory activity and its duration are actively controlled in humans.

Adult↗
PubMed1984

Brachial plexus injuries. Prognosis of postganglionic lesions.

One hundred ninety-eight cases of postganglionic brachial plexus injury with no signs of combined damage to preganglionic sites were treated conservatively. Among them, 50 whole, 32 upper and 13 lower type injuries were followed up for at least 2 years and the course of the progression of the individual muscle was observed. In the whole type, 44% showed good functional recovery, although some of them needed reconstructive surgery. Thirty-eight percent recovered usefully only in the muscles innervated by upper roots. Three out of 50 cases did not recover at all. There were no particular differences between the whole and upper types with regard to the recovery rate of the arm muscles. However, the upper type showed better functional recovery than the whole type. If the muscles innervated by upper roots could gain a strength of M1 before 9 months after injury, and muscles innervated by lower roots reached M1 before 12 months, their final results were more than M3. But muscles with a strength of less than M2 at 18 months did not recover to more than M3. The muscular power at 24 months was almost the same as the final results. About half of the postganglionic brachial plexus injuries which did not combine preganglionic lesions showed favorable spontaneous recovery with conservative treatment. At the exploration of the brachial plexus, if the somatosensory evoked potential test is positive, which negates the possibility of root avulsion, and also if there is no definite discontinuity of the postganglionic nerves, it should be treated conservatively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1984

Ossification of the posterior longitudinal ligament of the spine.

Ossification of the posterior longitudinal ligament of the spine (OPLL) is a newly recognized entity. As the incidence of this disease was exceptionally high in Japan, the Japanese Ministry of Public Health and Welfare instituted a special commission for the investigation of this perplexing disease; since 1975 this committee has performed an intensive study of 2100 patients with OPLL in Japan. An epidemiologic study was conducted by this group in Japan and in eastern Asiatic countries. Symptoms and disabilities caused by the disease were described. Roentgenographic findings were classified as continuous, segmental, mixed, or localized. OPLL at the thoracic and lumbar levels combined with ossification of the yellow ligament was described, and the risk of spinal cord damage as well as the importance of tomography and computerized tomographic scanning were stressed. No conclusions were reached concerning etiology, but common findings included a generalized hyperostotic tendency, a tendency for abnormal glucose metabolism, and low enteral calcium absorption. A relatively high hereditary occurrence was noted. Conservative and surgical treatment methods were described, with particular reference to spinal canal-widening operations.

Aged↗
PubMed1984

Recent trends in orthopaedics in Japan.

Orthopedic surgery in Japan has developed markedly since the end of the Second World War and at present many subspecialty study groups cover various aspects of basic orthopedic and clinical research. The main trends and topics of orthopedic surgery in Japan are presented as examples of recent or representative works.

Animals↗