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Biomedical subjects

T Watari

Publications and source records attributed to T Watari.

At least 55 records · Page 3Linked to original sources
PubMed1993

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↗
PubMed1993

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↗
PubMed1993

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↗
PubMed1993

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↗
PubMed1993

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↗
PubMed1993

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↗
PubMed1993

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↗
PubMed1993

[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↗
PubMed1992

Detection of enhancer repeats in the long terminal repeats of feline leukemia viruses from cats with spontaneous neoplastic and nonneoplastic diseases.

Enhancer duplication in the long terminal repeat of feline leukemia virus (FeLV) was examined in primary cells from naturally FeLV-infected cats with various neoplastic and nonneoplastic diseases using the polymerase chain reaction. In all cases, a 170-bp band, corresponding to a standard exogenous FeLV with one copy of enhancer, was detected. Repeated enhancer sequences were found in all 8 cases of thymic-form lymphosarcoma, in some cases of lymphosarcoma of other forms (3/8) and myeloid tumors (2/3), and in only 1 of 6 cases with nonneoplastic diseases. The copy number of FeLV proviruses with a repeated enhancer seemed higher than that of those with one copy of enhancer in 3 cases of thymic form lymphosarcoma. In 5 cases of thymic form lymphosarcoma and in 1 case of erythroleukemia, coexistent FeLVs with double and triple enhancers of different sizes were found. Of the enhancer elements, only the SV40 core binding site was found in all the enhancer direct repeats of these FeLVs. All the provirus clones with single and duplicated enhancer sequences from a single tumor showed mutations or deletions characteristic to that tumor, indicating that enhancer repeats may arise in individual animals after infection with a single virus clone. The present findings indicate that FeLV with enhancer repeats generated in the cat is associated with the induction of neoplastic diseases in natural conditions.

Animals↗
PubMed1992

Elevation of feline interleukin 6-like activity in feline immunodeficiency virus infection.

The involvement of feline interleukin 6 (IL-6)-like activity in polyclonal B-cell activation in feline immunodeficiency virus (FIV) infection was investigated by using the proliferative response of hybridoma cell clone B3B1. Cats spontaneously infected with FIV had hyperimmunoglobulinemia, as shown by increases in the percentage of gamma-globulin and the plasma IgG concentration and decrease in the albumin/globulin (A/G) ratio. Because IL-6 plays an essential role in the differentiation of activated B cells into Ig-secreting cells, we examined the effect of FIV infection on the plasma IL-6 level. Plasma IL-6-like activity was found to be significantly higher in FIV-infected cats than in healthy controls. Peripheral blood mononuclear cells (PBMC) isolated from FIV-infected cats and cultured without any exogenous activators of IL-6 production released more feline IL-6-like activity than cells from healthy controls. This phenomenon was mainly due to the increase in the production of IL-6 by adherent cells such as monocytes/macrophages, but also partly by nonadherent cells. These results indicate that elevation of feline IL-6-like activity is associated with FIV infection and that overproduction of IL-6 may contribute to the polyclonal B-cell activation seen in FIV infection.

Animals↗
PubMed1992

Altered surface antigen expression on peripheral blood mononuclear cells in cats infected with feline immunodeficiency virus.

Expression of CD4, CD8, IL-2 receptor alpha chain (IL-2R alpha), and MHC class II (MHC-II) on peripheral blood mononuclear cells were examined in cats infected with feline immunodeficiency virus (FIV). CD4/CD8 T cell ratio in FIV-infected cats was slightly decreased, as compared with that in specific-pathogen-free (SPF) cats. However, there was no statistical differences between them. The number of circulating IL-2R alpha+ cells in FIV-infected cats was higher than that in healthy cats, whereas induction of IL-2R alpha expression by concanavalin A (Con A) stimulation was depressed in FIV-infected cats. By using two-color cytofluorometry, Con A-induced enhancement of IL-2R alpha expression was found to be reduced in both CD4+ and CD8+ populations in PBMC from FIV-infected cats. The circulating MHC-II+ cells were also increased in FIV-infected cats. Furthermore, the induction of IL-2R alpha expression on PBMC after Con A-stimulation significantly depressed by FIV inoculation in vitro. These results suggest that FIV activates PBMC in vivo via direct and/or indirect mechanisms, leading to the unresponsive state of T cells to further stimuli in vitro.

Animals↗
PubMed1991

Exercise-induced phospholipid degradation in the equine skeletal muscle and erythrocytes.

To understand the pathogenesis of equine exercise-induced myopathies and hemolysis, changes of phospholipid peroxidation products in the equine middle gluteal muscle and erythrocytes following the high-speed treadmill exercise were studied. In the skeletal muscle, the peroxidized phosphatidylethanolamine (PE) were increased at 24 hours after the exercise. The malondialdehydes (MDAs) were also increased as the protein-bound MDAs following exercise. In the erythrocytes, the peroxidized PE were significantly decreased at 24 hours after the exercise. The protein-bound MDAs were significantly increased at 5 min after the exercise and returned to the base values at 24 hours after the exercise. These findings indicate that the PE is more susceptible to in vivo oxidative effects than the other phospholipid classes, and the accumulation of the protein-bound MDAs is considered to play some cytotoxic roles in the equine skeletal muscle and erythrocytes following exercise.

Animals↗
PubMed1990

Immune mediated skin lesions in a dog with autoimmune hemolytic anemia.

A canine case of Coombs' test positive and antinuclear antibody-negative hemolytic anemia was examined because of the development of skin lesions after 18 months treatment with prednisolone. Histopathological examination of biopsy specimens obtained from skin and oral mucosa revealed the acantholysis, edematous lesions of the stratum basale and mononuclear cell accumulation in the dermis. Deposits of immunoglobulin G and complement factor 3 were detected at the intercellular and dermoepidermal junction by the direct immunofluorescent test. From these results, the case was considered to be an autoimmune disease caused by distinct antibodies against different organs.

Anemia, Hemolytic, Autoimmune↗
PubMed1990

[Mixed type total anomalous pulmonary venous connection with the left upper pulmonary vein draining into the innominate vein and the other pulmonary veins into the coronary sinus--surgical correction in 2 infants].

Surgical repair of the mixed type total anomalous pulmonary venous connection (TAPVC) in infancy is known to be difficult. In this report, experience of two infants with the mixed type TAPVC with the left upper pulmonary vein (PV) draining into the innominate vein and the other PVs into the coronary sinus (IIa + Ia (left upper PV) type TAPVC) is presented. In such cases, it has been said that the left upper PV should be left uncorrected, but we anastomosed the left upper PV, that was thin and fragile, to the left atrium and corrected all the other anomalous PVs. The results were satisfactory. Recently, Extra-Corporeal circulation, microsurgery, and other techniques have significantly progressed, so we consider that the anastomosis is possible in most infants with IIa + Ia (left upper PV) type TAPVC. In the future, careful clinical follow-up and repeat catheterizations should be done, and we'd like to examine the long term patency of the left upper PV.

Anastomosis, Surgical↗
PubMed1990

[Subtotal cor triatrium with left partial anomalous pulmonary venous connection--successful correction in an infant].

This report concerns a 10-month-old infant with subtotal cor triatrium associated with left sided partial anomalous pulmonary venous connection to the innominate vein. In the operation, we found that the fossa ovalis existed between the right atrium and the accessory atrial chamber. We performed a radical operation which consisted of a resection of the intra-left-atrial diaphragma and a anastomosis of the vertical vein to the left atrium. His postoperative course was uneventful, and the result was satisfactory. We have presented and reviewed our case, and then discussed the embryogenesis and the hemodynamics. To our knowledge, our case represents the first successful surgical repair of this combination of defects in Japan.

Brachiocephalic Veins↗
PubMed1989

Evaluation and comparison of the axis of the interventricular septum by two-dimensional echocardiography among the patients with congenital heart disease (ASD, VSD, PDA, PS, TOF) and the normal subjects.

A two-dimensional echocardiographic method was applied to determine the axis of interventricular septum (IVS) in the horizontal plane. This study comprised 19 subjects with ASD, 15 subjects with VSD(I), 20 subjects with VSD(II), 13 subjects with PDA, 16 subjects with PS, 15 subjects with TOF and 99 normal children. Parasternal left ventricular short axis view was taken, and the IVS was recorded by using the polaroid prints. The axis of the IVS in the horizontal plane was measured from the recorded polaroid prints. The value of the angularity of IVS (IVS-A) expressed as means +/- one standard deviation (mean +/- 1S.D.) is 52.0 +/- 13.7 degrees in ASD, 42.2 +/- 9.7 degrees in VSD(I), 43.7 +/- 9.1 degrees in VSD(II), 41.6 +/- 11.4 degrees in PDA, 38.6 +/- 10.9 degrees in PS, 61.0 +/- 8.5 degrees in TOF and 40.1 +/- 8.2 degrees in normal controls. There was no significant difference among VSD(I), VSD(II), PDA, PS and normal controls but a highly significant difference was noted in ASD and TOF as compared to other groups. The correlation coefficient of the IVS-A with (1) the Qp/Qs ratio, (2) the magnitude of shunt (%), (3) the right ventricular pressure and (4) the hematocrits were evaluated among the patient groups. The IVS-A of ASD and VSD(II) had a good correlation with the shunt flow (r = 0.921 and 0.88 respectively) and/or the Qp/Qs ratio (r = 0.782 and 0.955 respectively); while that of VSD(I), PDA, and PS had a good correlation with the right ventricular pressure (r = 0.956, 0.953, 0.842 respectively) and that of TOF was mostly concerned with the hematocrits (r = 0.911). Besides, the IVS-A in each severe CHD subgroup was significantly (p less than 0.005 or a better value) higher than the normal control group. Thus an alternative method was validated for evaluating the severity of congenital heart disease by using a noninvasive two-dimensional echocardiographic technique.

Child↗