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Biomedical subjects

T Watari

Publications and source records attributed to T Watari.

At least 73 records · Page 4Linked to original sources

Molecular cloning of equine interleukin-1 alpha and -beta cDNAs.

Equine interleukin-1 alpha (IL-1 alpha) and IL-1 beta were molecularly cloned to establish a basis for research on inflammatory and immune responses in the horse. Equine peripheral blood mononuclear cells (PBMC) were stimulated with lipopolysaccharide (LPS), and cDNA clones of equine IL-1 alpha and IL-1 beta covering the whole coding sequences were isolated from them. These equine IL-1 alpha and IL-1 beta clones contained open reading frames encoding 271 and 269 amino acids, respectively. The deduced amino acid sequence of equine IL-1 alpha showed 71.6% and 60.2% similarity with that of human and murine IL-1 alpha, respectively. Similarly, the amino acid sequence of equine IL-1 beta showed 66.7% and 61.8% similarity with that of human and murine IL-1 beta, respectively. In both equine IL-1 alpha and IL-1 beta, amino acids at miristoylation sites were well conserved. Dot blot analysis indicated that the expression of IL-1 beta was predominant to that of IL-1 alpha in equine PBMC stimulated with LPS or phorbol myristate acetate.

Amino Acid Sequence↗

Gammopathy with two M-components in a dog with IgA-type multiple myeloma.

A 12-year neutered male mixed-breed dog was referred to hospital for evaluation of chronic diarrhea. Cellulose acetate electrophoresis of its serum revealed two monoclonal peaks in the gamma-globulin fraction. On immunoelectrophoretic analysis, the two monoclonal peaks in the gamma-globulin region were strongly precipitated with anti-dog IgA serum. On sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis, the fractions corresponding to these two peaks were shown to be dimer and trimer or tetramer of immunoglobulin consisting of heavy and light chains. These results indicated that the studied dog had gammopathy with two M-components with dimer and trimer or tetramer of IgA. Accumulations of large amounts of these immunoglobulins with very high molecular weight in the serum were concluded to induce the hyperviscosity syndrome in this dog in the terminal stage.

Animals↗

Erythroleukemia in two cats naturally infected with feline leukemia virus in the same household.

Erythroleukemia was observed in two unrelated cats infected with feline leukemia virus (FeLV) from the same household. Case 1, a 1-year-old neutered male cat developed erythroleukemia (M6) after a diagnosis of myelodysplastic syndrome (MDS-Er) on the criteria of FAB classification of acute leukemias. Case 2, a 1-year-old neutered female cat, which had close contact with Case 1, also developed erythroleukemia (M6Er). In both cases, marked proliferation of erythroid progenitor cells with disproportionally large numbers of immature forms was observed in the bone marrow. In Case 1, neoplastic proliferation of myeloid cells in the bone marrow was also noted at the terminal stage. Combination chemotherapy with daunomycin was partially effective for treatment of these erythroid neoplasias, but did not induce complete remission. Southern blot analysis using exogenous FeLV-specific probes indicated the clonal origin of these hematopoietic tumor cells. Furthermore, the erythroid and myeloid tumor cells in Case 1 were shown to be derived from independent transformed clones. A variant FeLV was shown to be integrated into the tumor cells in Case 1, while a full-length FeLV was found in both cases. Because these erythroid neoplastic diseases occurred in two unrelated cats kept in the same household and these diseases are rare, they may both have been associated with the same FeLV strain.

Animals↗

Two forms of the mb-1 gene transcript in cattle.

We have isolated two forms of bovine mb-1 cDNA clones from a cDNA library of a bovine leukemia virus-infected B-lymphoblastoid cell line. The nucleotide sequence analysis of these cDNA clones indicated that the shorter form of cDNA had a 119-bp deletion that precisely corresponded to the exon III domain of the human and murine mb-1 genes. This deletion would result in a frame shift, producing a premature translation termination leading to the lack of the entire transmembrane and cytoplasmic domains of the normal mb-1 molecule. The existence of two species of mRNA transcripts corresponding to the isolated cDNA clones were confirmed in all of the bovine leukemia cell lines examined by PCR with the use of the primers flanking the deletion site. Furthermore, the shorter mb-1 transcript was induced in normal PBMC by stimulation with mitogens or culture supernatants of bovine leukemia virus-infected cell lines. These findings indicate the presence of the truncated mb-1 gene product which may have some function different from that of the normal mb-1 gene product.

Alternative Splicing↗

Cloning of feline p53 tumor-suppressor gene and its aberration in hematopoietic tumors.

Alterations of the p53 tumor-suppressor gene have been observed in a variety of human and mouse tumors. For investigation of the role of this gene in tumors of cats, feline p53 cDNA was molecularly cloned from a feline lymph-node cDNA library. The cloned cDNA (FF53) contained the whole open reading frame of p53 gene encoding 386 amino acids. The amino-acid sequence of the feline p53 gene showed 82.1% and 74.9% similarities with those of the human and mouse counterparts, respectively, and had structural characteristics in common with the p53 genes of several other species. Aberrations of the p53 gene were investigated by RT-PCR and single-strand conformation polymorphism analyses. Of 10 primary hematopoietic tumors and 3 lymphoma cell lines examined, one lymphoma and one lymphoma cell line had a point mutation of the p53 gene, resulting in single amino-acid substitutions.

Amino Acid Sequence↗

Induction of apoptosis in a T lymphoblastoid cell line infected with feline immunodeficiency virus.

The mechanism of cell death induced by feline immunodeficiency virus (FIV) infection was investigated in an interleukin 2(IL-2)-dependent T-lymphoblastoid cell line (MYA-1). DNA extracted from FIV-infected MYA-1 cells showed a ladder of nucleosomal DNA, indicating that the cytopathic effect (CPE) observed in these cells was due to apoptosis. Infection of MYA-1 cells with FIV was associated with suppression of the proliferative response of the cells to exogenous IL-2 prior to DNA fragmentation. These findings suggest that FIV-induced CPE in these T-lymphoblastoid cells is associated with apoptosis possibly due to a defect in the IL-2 signal transduction pathway.

Animals↗

[123I-MIBG scintigraphy in patients with neuroblastoma].

123I-meta-iodobenzylguanidine (123I-MIBG) scintigraphy was performed in 9 patients who were suspected to have a neuroblastoma. Positive images were obtained in 4 patients who had a neuroblastoma. Negative images were obtained in 5 patients who did not have a neuroblastoma. 123I-MIBG scan revealed an active tumor in a patient who had received intensive chemotherapy for the recurrence of neuroblastoma with returning of all tumor markers including neuron specific enolase to normal range. This indicated that 123I-MIBG scan can be used to assess the effect of therapy.

3-Iodobenzylguanidine↗

Accumulation of thallium-201 in medullary thyroid cancer with negative serum calcitonin and carcinoembryonic antigens: a case report.

Thallium-201 (201Tl) scintigraphy has been used in the evaluation of thyroid neoplasm. Even though several authors have shown the usefulness of 201Tl scintigraphy in evaluating recurrent tumor of medullary thyroid cancer, the number of reports on primary thyroid cancer is limited. We presented a case of medullary thyroid cancer which showed 201Tl uptake in both early and delayed scans. This case had normal serum levels of calcitonin and carcinoembryonic antigen.

Calcitonin↗

Thallium-201 scintigraphy in the evaluation of thyroid nodules. A retrospective study of 246 cases.

There is controversy at present concerning the value of Tl-201 scintigraphy in the diagnosis of malignant thyroid nodules. The usefulness of Tl-201 scintigraphy in differentiating benign from malignant nodules was evaluated retrospectively. Tl-201 scintigraphy was performed in 101 patients with malignant thyroid nodules and in 145 with benign nodules. Early and late images were examined to compare Tl-201 uptake in nodules and normal thyroid tissue. The visual method resulted in 74% of sensitivity and 58% of specificity for the detection of malignant nodules including incidental cancers. Excluding incidental cancers, 87% sensitivity and 58% specificity were achieved. Small cancers, including incidental cancers less than 1.5 cm in diameter and small number of viable tumor cells in a large tumor also gave negative Tl-201 scintigraphic results. Forty-two percent of the benign lesions gave false positive Tl-201 scintigraphic results. It is concluded that Tl-201 scintigraphy is a sensitive and useful method to use in diagnosing malignant thyroid nodules, but is not a specific method even with late scan, for the detection of malignant thyroid nodules.

Adenocarcinoma, Follicular↗

Detection of feline immunodeficiency virus proviral DNA in feline peripheral blood mononuclear cells by the nested two-step polymerase chain reaction.

The polymerase chain reaction (PCR) was applied to detect feline immunodeficiency virus (FIV) proviral DNAs in primary peripheral blood mononuclear cells (PBMC). Suitable conditions for PCR amplification were examined to obtain highly sensitive and specific results by simple staining in agarose gel. Specific amplification of FIV proviral DNA in PBMC DNA of FIV-infected cats was achieved by a nested two-step PCR that amplified the DNA first with outer primers and then with inner primers nested within the first primers. PCR amplification using different primers indicated that those based on the gag sequence of the FIV/TM2 strain isolated in Japan were suitable for the detection of FIV genomes in naturally infected Japanese pet cats. By the nested two-step PCR with mixed gag primers of TM2 and Petaluma, isolated in the USA, we could detect FIV genomes in all 11 primary PBMC samples from FIV-seropositive cats tested. The PCR protocol developed here is sensitive and specific for molecular detection of FIV infection in cats.

Animals↗

Apoptosis induced by tumor necrosis factor in cells chronically infected with feline immunodeficiency virus.

Tumor necrosis factor alpha (TNF-alpha) induced morphologic changes such as chromatin condensation and cell shrinkage in a feline fibroblastic cell line (CRFK) chronically infected with feline immunodeficiency virus (FIV) but not in uninfected CRFK cells. DNA extracted from TNF-alpha-treated CRFK cells infected with FIV showed a ladder of nucleosomal DNA, indicating that this cytocidal effect by TNF-alpha was due to programmed cell death, or apoptosis. These findings may have implications for understanding the pathogenesis of FIV infection and for the design of specific therapeutic strategies for AIDS in humans as well as cats.

Animals↗

Malignant histiocytosis with multiple skin lesions in a dog.

A 7-year-old male Yorkshire terrier was examined for multiple plaques and nodules on the skin. The clinical, cytological and histopathological features indicated a malignant histiocytosis. Cytoreductive chemotherapy produced moderate clinical improvement, but died at day 90 after the first admission. Pathological examination revealed the neoplastic histiocytes in the skin as well as in the myocardium and skeletal muscles.

Animals↗

Rearrangements of immunoglobulin and T-cell receptor genes in canine lymphoma/leukemia cells.

Tumor cells from 15 canine lymphoma/leukemia cases were examined for genetic rearrangements of immunoglobulin and T-cell receptor (TCR) genes in parallel with cell surface antigens. Ten of these 15 cases showed rearrangements of the immunoglobulin heavy chain (IgH) gene, while 4 cases displayed TCR beta-chain gene rearrangements on Southern blot analysis. All the cases with IgH gene rearrangements had multicentric form lymphoma, and 6 of the 10 cases were cell surface immunoglobulin-positive. On the other hand, the cases with TCR gene rearrangements included atypical lymphoma/leukemia cases, and 3 of the 4 cases were Thy-1 antigen-positive. Although the tumor cell lineage of a considerable number of lymphoma/leukemia cases could not be determined by phenotypic analysis, examination of the IgH and TCR gene rearrangements disclosed the lineages of 14 of 15 cases. Genetic analysis demonstrated that the tumor cells in most canine multicentric lymphomas were formed by clonal expansion of B-lymphocyte. These findings show that studies on the rearrangements of immunoglobulin and TCR genes are very useful for understanding the cellular origin, clonality and hierarchy of canine lymphoma/leukemia cells.

Animals↗

Molecular cloning and chromosomal mapping of feline p53 tumor suppressor gene.

Alterations of the p53 tumor suppressor gene have been observed in a variety of human and mouse tumors. For investigation of the role of this gene in tumors in cats, feline p53 cDNA was molecularly cloned by PCR amplifications using primers based on the sequences conserved among several species. The cloned cDNA appeared to cover approximately 90% of the open reading frame of the feline p53 gene and had characteristic structures in common with the p53 genes of several other species. The amino acid sequence similarities of the feline p53 with the human, mouse, rat and chicken counterparts were 82.9%, 75.6%, 76.5% and 57.2% respectively. Moreover, using a panel of feline x rodent somatic cell hybrids, the feline p53 gene was assigned to feline chromosome E1. These data will be useful for determining the role of the p53 tumor suppressor gene in feline tumors.

Amino Acid Sequence↗

Establishment and characterization of a feline large granular lymphoma cell line expressing interleukin 2 receptor alpha-chain.

A feline large granular lymphocyte (LGL) cell line was established from a cat with an alimentary-form lymphoma. This cell line, designated as FGL, had many large azurophilic granules in the cytoplasm, which were typical to LGL cells. Proviral genome of feline leukemia virus was detected in the chromosomal DNA of FGL cells, and reverse transcriptase activity was also demonstrated in the culture supernatant. Furthermore, we found expression of interleukin 2 (IL-2) receptor alpha-chain on the cell surface of FGL and its natural killer activity against human erythroblastic leukemia cell line, K562.

Animals↗

Molecular cloning of feline interleukin-6 cDNA.

We cloned and sequenced feline interleukin-6 (IL-6) cDNA using the polymerase chain reaction (PCR) technique to obtain information on regulation of the IL-6 gene. The feline IL-6 cDNA cloned in this study was 752 bp long and included all the amino acid coding region. At the amino acid level, the overall identities of the feline IL-6 to the human and murine counterparts were 58% and 39%, respectively. After transfection with the expression vector containing the cDNA, CRFK cells produced biologically active proteins that showed hybridoma growth promoting activity.

Amino Acid Sequence↗

[Evaluation of renal uptake on 111InCl3 bone marrow scintigraphy in patients with aplastic anemia and myelodysplastic syndrome].

High renal uptake on bone marrow scan with indium-111 chloride is often shown in patients with bone marrow abnormality. We evaluated the renal uptake on bone marrow scan in 27 cases with aplastic anemia, 20 cases with myelodysplastic syndrome (MDS) and 10 cases with other diseases. The high renal uptake was observed in patients not only with aplastic anemia but also with MDS. The renal uptake correlated with blood transfusion units, unsaturated iron binding capacity (UIBC), blood pool imaging and bone marrow uptake. The renal uptake correlated with UIBC better than with the blood transfusion units. Following mechanism of the renal uptake is proposed that frequent blood transfusion makes low UIBC, and the low UIBC causes the failure to chelate indium with transferrin. The fast blood clearance of un-chelated indium via kidneys is followed. Hypoplastic bone marrow may also play an important role for the high renal uptake because all patients with the high renal uptake could not be explained by above mentioned mechanisms. Caution should be paid to the scans with the high renal uptake because both aplastic anemia and MDS patients show the high renal uptake.

Anemia, Aplastic↗