PubMed Health⌕ Search

Biomedical subjects

R Nishimura

Publications and source records attributed to R Nishimura.

At least 91 records · Page 5Linked to original sources
PubMed1999

Selectively induced high MRP gene expression in multidrug-resistant human HL60 leukemia cells.

A subclone HL60/DOX was selected from a human leukemic HL60 cell line for resistance to doxorubicin (DOX) by exposure to stepwise increasing concentrations of the drug and coexposure to a potential P-glycoprotein (P-gp) inhibitor, cepharanthine (a biscoclaurine alkaloid). Compared with the parent HL60 cells, the HL60/DOX cells were 13.0-fold more resistant to DOX and showed multidrug-resistant (MDR) phenotype characterized by 4.6-fold, 2.3-fold, and 5.7-fold cross-resistance to vincristine, pirarubicin, and etoposide, respectively, but no cross-resistance to alkylating agent, cisplatin. Immunocytochemical analyses using the specific monoclonal antibody, MRPr1, and quantitative analyses using a competitive reverse transcription-polymerase chain reaction (CRT-PCR) confirmed overexpression of MRP gene products (about 8-fold determined by CRT-PCR) in this resistant clone. The P-gp expression was not detectable by the monoclonal antibody, C219, in the HL60/DOX cells, and that was consistent with extremely low levels of mdr1 mRNA expression determined by CRT-PCR in this clone. Drug accumulation and efflux studies demonstrated the significantly increased efflux rate of DOX compared to the parent HL60 cells. This enhancement of DOX efflux was reversed by the addition of 10 microM verapamil. To investigate the additional underlying mechanisms contributing to MDR phenotype in the HL60/DOX cells, the levels of DNA topoisomerases (Topo) including Topo I, Topo IIalpha, and Topo IIbeta, and gamma-glutamylcystein synthetase (y-GCS) expression were determined using CRT-PCR techniques. Normal expression of each enzyme at the transcriptional level was demonstrated in this resistant clone. Southern blot analysis of the gene organization in the HL60/DOX cells revealed the amplification of MRP gene. These results indicate that alteration of the drug accumulation from enhanced efflux appears to be a major mechanism(s) of MDR phenotype and attributable to high levels of MRP expression in the HL60/DOX cells. Overexpression of MRP in this clone is regulated by the genomic amplification of DNA and increased levels of the MRP mRNA, independently with the normal expression of Topo I, Topo IIalpha, Topo IIbeta, or gamma-GCS.

ATP-Binding Cassette Transporters↗
PubMed1999

Levels of soluble FasL and FasL gene expression during the development of graft-versus-host disease in DLT-treated patients.

Three patients with different clinical symptoms of graft-versus-host disease (GVHD) who had received donor lymphocyte transfusion (DLT) for the treatment of relapsed leukaemia after an allogeneic bone marrow transplantation (BMT) from HLA-matched sibling donors were analysed for the presence of soluble FasL (sFasL) in the sera and for the expression of the Fas ligand (FasL) gene in the peripheral blood mononuclear cells (PBMNC). Two patients who demonstrated liver damage with increased levels of serum bilirubin showed significantly increased levels of serum sFasL. The increase in the sFasL level was observed prior to the increase in the bilirubin during the clinical courses of both patients. The high dose of methyl predonisolone administered to one of these patients greatly reduced the levels of sFasL in the serum. The bilirubin levels were also reduced thereafter. The third patient (without liver damage) did not show any increase in the serum sFasL level. The expression of the FasL gene in the PBMNC of these three patients was examined. All three patients showed increased levels of the FasL gene expression during their clinical courses. However, only one patient showed a parallel alteration of FasL gene expression with sFasL in the serum. These cases provide evidence that the Fas/FasL system is closely associated with human GVHD, especially in the development of liver GVHD.

Adult↗
PubMed1999

Comparison of sedative effects of medetomidine-midazolam, acepromazine-butorphanol and midazolam-butorphanol in dogs.

Sedative effects of medetomidine at 20 micrograms/kg and midazolam at 0.3 mg/kg (MM), acepromazine at 0.05 mg/kg and butorphanol at 0.2 mg/kg (AB), and midazolam at 0.1 mg/kg and butorphanol at 0.2 mg/kg (MB) were compared in dogs. All dogs given MM were laterally recumbent within 11 min of the administration of the drugs, and this combination also induced deep sedation accompanied by analgesia and muscle relaxation, and strongly depressed arousal reactions to external stimuli. However, undesirable effects such as bradycardia and decreased respiration were also observed in dogs given MM. Six of seven dogs given AB were laterally recumbent within 16 min of the administration of the drugs, and this combination induced relatively deep sedation but only a mild depression of arousal reactions to external stimuli. MB induced mild or moderate sedation with relatively large differences in effects among individuals. The recovery from sedation in each group was smooth and total recovery times were not significantly different.

Acepromazine↗
PubMed1999

Effects of permanent tracheostomy on respiratory reflexes to lung inflation and capsaicin in sevoflurane-anaesthetized dogs.

The aim of this study was to evaluate the effects of permanent tracheostomy on baseline breathing, reflex responses to lung hyperinflation (1.5 kPa) and right atrial capsaicin injection (5 micrograms/kg) before and at 3 and 5 weeks after tracheostomy in sevoflurane-anaesthetized spontaneously breathing dogs (n = 6). In all observation periods, apnoeic responses, represented by an increase in expiration time after lung inflation and right atrial capsaicin injection, were consistently observed to be a result of the Hering-Breuer inflation reflex and the pulmonary C-fibre chemoreflex. Investigation at 3 and 5 weeks after tracheostomy revealed no significant evidence of changing baseline breathing pattern or reflex responses to lung inflation and capsaicin in any ventilatory variable (inspiration time, expiration time, tidal volume, expired ventilation, and end-tidal PCO2) from levels recorded before surgery. These results indicate that permanent tracheostomy, at least up to 5 weeks, does not affect the baseline breathing pattern, the Hering-Breuer inflation reflex or the pulmonary C-fibre chemoreflex. Further, our investigation provides a useful canine model in respiratory physiology.

Anesthetics, Inhalation↗
PubMed1999

Comparison of cardiopulmonary effects of medetomidine-midazolam, acepromazine-butorphanol and midazolam-butorphanol in dogs.

Cardiopulmonary effects of intramuscular administration of medetomidine-midazolam at 20 micrograms/kg and 0.3 mg/kg, respectively (MM), acepromazine-butorphanol at 0.05 and 0.2 mg/kg, respectively (AB), and midazolam-butorphanol at 0.1 and 0.2 mg/kg, respectively (MB), were compared in dogs. MM produced relatively large cardiovascular changes such as bradycardia, hypertension, a decrease in cardiac output and vasoconstriction. AB produced moderate cardiovascular changes. Hypotension was observed within 5 min after administration and the mean arterial blood pressure decreased by 30% at 20 min after administration. MB produced decreases in arterial blood pressure and cardiac output, but these changes were small and, of the combinations evaluated in this study, MB had the mildest cardiovascular effects.

Acepromazine↗
PubMed1999

Microsatellite analysis of childhood leukemia: correlation of 9p and 12p chromosome abnormalities with expression of related genes.

BACKGROUND: Loss of heterozygosity (LOH) and DNA replication error (RER) have been thought to be involved in carcinogenesis, but have not been investigated in childhood leukemia and lymphoma. METHODS: Eighty samples from 65 patients with childhood leukemia and lymphoma were examined using seven different microsatellite markers for RER analysis. Additionally, LOH in two chromosome regions (9p and 12p) was investigated. Furthermore, expression of the TEL, TEL/AML1 and p27(KIP1) genes on 12p and the p16 gene on 9p were detected by reverse transcriptase polymerase chain reaction. RESULTS: Replication errors were detected in 5/65 patients (7.7%). Most (4/5 patients) RER were preferentially located in the 9p and 12p regions. There were two patients who had DNA abnormalities in both 9p and 12p, one with common acute lymphoblastic leukemia (ALL) showed 9p LOH and the TEL/AML1 fusion gene on 12p and the other with common ALL and 12p RER had diminished expression of both the p27(KIP1) gene on 12p and the p16 gene on 9p. CONCLUSIONS: Combined DNA alterations on 9p and 12p, involving LOH, RER and/or gene mutation and chromosomal translocation, were found in childhood acute leukemia, especially in common ALL.

Child↗
PubMed1999

Effects of volatile anesthetics on the activity of laryngeal 'drive' receptors in anesthetized dogs.

Effects of halothane, isoflurane and sevoflurane on laryngeal drive receptor activity were studied in the afferent activity of the superior laryngeal nerve in anesthetized spontaneously breathing dogs. Of 40 single units recorded, most of them (65%) responded to the volatile anesthetics applied to the isolated larynx at a concentration of 5%. The exposure to the anesthetics resulted in either an inspiratory increase (15%), both inspiratory and expiratory decrease (54%), or both inspiratory increase and expiratory decrease (31%) responses. The average discharge frequency of the receptors tended to be decreased on inhalation of the anesthetics, where significant decreases were observed in both respiratory phases for halothane and at expiration for isoflurane, but in neither respiratory phase for sevoflurane. These results support an advantage of sevoflurane over halothane and isoflurane for induction of anesthesia to minimize the influence of the activity of laryngeal drive receptors on the breathing pattern and airway stability.

Anesthetics, Inhalation↗
PubMed1999

Measurement of telomerase activity in dog tumors.

Telomeres are specific structures present at the end of liner chromosomes. DNA polymerase can not synthesize the end of liner DNA and, as a result, the telomeres become progressively shortened by successive cell divisions. To overcome the end replication problem, telomerase adds new telomeric sequences to the end of chromosomal DNA. The enzyme activity is undetectable in most normal human adult somatic cells, in which shortening of the telomere is thought to limit the somatic-cell life span. In contrast to normal somatic cells, many human tumors possess telomerase activity. The present study looked at whether telomerase activity might serve as a marker for canine tumors. Telomerase activity was measured using the telomeric repeat amplification protocol assay. Normal dog somatic tissues showed little or no telomerase activity, while normal testis exhibited a high level of telomerase activity. We measured telomerase activity in tumor samples from 45 dogs; 21 mammary gland tumors, 16 tumors developed in the skin and oral cavity, 7 vascular tumors and 1 Sertoli cell tumor. Greater than 95% of the tumor samples contained telomerase activity (3-924 U/2 micrograms protein). The results obtained in this study indicated that telomerase should be a useful diagnostic marker for a variety of dog tumors, and it may serve as a target for antitumor chemotherapy.

Adenocarcinoma↗
PubMed1999

1,25-Dihydroxyvitamin D3, recombinant human transforming growth factor-beta 1, and recombinant human bone morphogenetic protein-2 induce in vitro differentiation of canine osteosarcoma cells.

Effects of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), recombinant human transforming growth factor (rhTGF)-beta 1 and recombinant human bone morphogenetic protein (rhBMP)-2 on differentiation in four different canine osteosarcoma cell lines (POS53B, 53C, 53D and 14A) were examined using markers specifically expressed by phenotypic osteoblasts. 1,25(OH)2D3 increased alkaline phosphatase (ALP) activity in one cell line, osteocalcin production in two lines and type I collagen production in three lines. RhTGF-beta 1 increased ALP activity in one clonal cell, osteocalcin production in one clonal cell and type I collagen production in two clonal cells. RhBMP-2 increased ALP activity in all clonal cells, osteocalcin production in two clonal cells and type I collagen production in three clonal cells. Thus, these agents induced differentiation in osteosarcoma cells at different efficacies. Electron microscopic study revealed that these agents increased cellular activity in all cell lines with no evidence of degeneration of cell organelle by drug cytotoxicity. In some cultures treated with either 1,25(OH)2D3 or rhBMP-2, apoptotic cells were observed. Based on the change in markers, rhBMP-2 and 1,25(OH)2D3 seemed to be more effective than rhTGF-beta 1. These agents are potential inducers of apoptosis.

Alkaline Phosphatase↗
PubMed1999

The posterior nasal nerve plays an important role on cardiopulmonary reflexes to nasal application of capsaicin, distilled water and l-menthol in anesthetized dogs.

The sensory innervation of the cardiopulmonary reflexes to nasal application of capsaicin (CAPS), distilled water (DW) and l-menthol (LM) was studied in anesthetized dogs breathing through tracheostomy. A marked cardiopulmonary reflex was observed by CAPS and DW into the nasal cavity, while a prolongation of expiration was induced by LM. All these reflexes were significantly decreased by bilateral section of the posterior nasal nerve (PNN) and completely abolished by topical nasal anesthesia with lidocaine. Responses of the whole nerve activity of the PNN to these substances corresponded to the magnitude of the reflexes. These results indicate that PNN afferents play an important role on the reflex elicitation of the noxious, water and cold stimuli from the nasal cavity.

Administration, Intranasal↗
PubMed1999

Nasal sensory receptors responding to capsaicin, water and tactile stimuli in sevoflurane-anesthetized dogs.

Responses of nasal receptors to capsaicin and water were studied from afferent recordings of the posterior nasal nerve (PNN) in 12 anesthetized dogs. Out of 12 non-respiration-modulated nasal receptors, 7 responded only to capsaicin, 3 responded to both water and capsaicin, and 2 to neither of them. All the fibers showed a rapid adaptation to mechanical probing of the nasal mucosa. These results indicate that the presence of sensory receptors responding to capsaicin and water are involved in PNN afferents of the dog.

Anesthesia, Inhalation↗
PubMed1999

Generalized dermal dysplasia and perifollicular mucinosis in a dog.

Multiple nodules 0.5-2 cm in diameter were observed in the dorsal skin of a 1.5-year-old male castrated mongrel dog. Histopathologically, perifollicular mucinosis and clusters of small cells with a dense, round nucleus and scant cytoplasm were seen. The small cell clusters were present in the dermis near the epidermal basal layer or surrounding hair follicles. These cells demonstrated different staining patterns for vimentin and keratin depending on their location; ultrastructurally, they were poor in organelles and connected to each other with cytoplasmic projections. These findings suggested that they were immature epithelial cells. The case was diagnosed as generalized dermal dysplasia and perifollicular mucinosis.

Animals↗
PubMed1999

MR imaging of squamous cell carcinoma of the floor of the mouth. Appearance of the sublingual and submandibular glands.

PURPOSE: To determine the diagnostic value of MR imaging for tumors of the floor of the mouth and the effects of the tumors on the sublingual and submandibular glands. MATERIAL AND METHODS: Thirty-seven patients with proven squamous cell carcinoma of the floor of the mouth underwent MR imaging, including unenhanced T1-weighted, T2-weighted, dynamic, and contrast-enhanced T1-weighted images. The appearance of the tumor and the sublingual and submandibular glands was assessed qualitatively and quantitatively. RESULTS: All tumors demonstrated replacement of the normal signal intensity in the adjacent sublingual gland. Twenty-one patients (57%) had abnormal signal intensity of the submandibular gland without tumor invasion, presumably secondary to submandibular duct obstruction by the tumor. Unenhanced T1-weighted images provided high contrast between tumor and sublingual gland. Tumors limited within the gland were well detected on unenhanced T1-weighted images. Large tumors extending beyond the gland were well delineated on dynamic images, but no better than on T2-weighted images. CONCLUSION: At MR imaging for tumor of the floor of the mouth, one must carefully evaluate the appearance of the sublingual and submandibular glands. Contrast-enhanced studies are unnecessary when the tumor is limited within the sublingual gland on precontrast MR images.

Adult↗
PubMed1999

[Baroreflex function in patients with Binswanger's encephalopathy].

To analyze the baroreflex function in Binswanger's encephalopathy, we performed head-up tilting (HUT) in three steps (20 degrees, 40 degrees, and 60 degrees for 5 min.) and evaluated changes in blood pressure, heart rate, plasma noradrenaline (NA) and plasma arginine vasopressin (AVP) in 15 patients with Binswanger's encephalopathy (mean age; 70.1 +/- 5.6 years), 10 patients with lacunar infarction (mean age; 72.3 +/- 6.3 years) and eight normal controls (mean age; 69.0 +/- 4.3 years). In the patients with Binswanger's encephalopathy, the resting blood pressure was higher than that in the other two groups and the blood pressure decreased by more than 20 mmHg in five cases and by 15 mmHg in one case after HUT. In all groups, the heart rates and NA increased significantly after HUT, but the AVP levels were not significantly increased after HUT in the patients with Binswanger's encephalopathy. In conclusion, central baroreceptor pathways involved in the function of AVP release, brainstem-hypothalamic pathways are impaired in Binswanger's encephalopathy.

Aged↗
PubMed1998

Optimal image resolution for digital storage of radiotherapy-planning images.

PURPOSE: To evaluate the quality of digitized radiation-planning images at different resolution and to determine the optimal resolution for digital storage. METHODS AND MATERIALS: Twenty-five planning films were scanned and digitized using a film scanner at a resolution of 72 dots per inch (dpi) with 8-bit depth. The resolution of scanned images was reduced to 48, 36, 24, and 18 dpi using computer software. Image qualities of these five images (72, 48, 36, 24, and 18 dpi) were evaluated and given scores (4 = excellent; 3 = good; 2 = fair; and 1 = poor) by three radiation oncologists. An image data compression algorithm by the Joint Photographic Experts Group (JPEG) (not reversible and some information will be lost) was also evaluated. RESULTS: The scores of digitized images with 72, 48, 36, 24, and 17 dpi resolution were 3.8 +/- 0.3, 3.5 +/- 0.3, 3.3 +/- 0.5, 2.7 +/- 0.5, and 1.6 +/- 0.3, respectively. The quality of 36-dpi images were definitely worse compared to 72-dpi images, but were good enough as planning films. Digitized planning images with 72- and 36-dpi resolution requires about 800 and 200 KBytes, respectively. The JPEG compression algorithm produces little degradation in 36-dpi images at compression ratios of 5:1. CONCLUSION: The quality of digitized images with 36-dpi resolution was good enough as radiation-planning images and required 200 KBytes/image.

Humans↗
PubMed1998

Cadherin-6 mediates the heterotypic interactions between the hemopoietic osteoclast cell lineage and stromal cells in a murine model of osteoclast differentiation.

Osteoclasts are multinucleated cells of hemopoietic origin that are responsible for bone resorption during physiological bone remodeling and in a variety of bone diseases. Osteoclast development requires direct heterotypic cell-cell interactions of the hemopoietic osteoclast precursors with the neighboring osteoblast/stromal cells. However, the molecular mechanisms underlying these heterotypic interactions are poorly understood. We isolated cadherin-6 isoform, denoted cadherin-6/2 from a cDNA library of human osteoclast-like cells. The isolated cadherin-6/2 is 3,423 bp in size consisting of an open reading frame of 2,115 bp, which encodes 705 amino acids. This isoform lacks 85 amino acids between positions 333 and 418 and contains 9 different amino acids in the extracellular domain compared with the previously described cadherin-6. The human osteoclast-like cells also expressed another isoform denoted cadherin-6/1 together with the cadherin-6. Introduction of cadherin-6/2 into L-cells that showed no cell-cell contact caused evident morphological changes accompanied with tight cell-cell association, indicating the cadherin-6/2 we isolated here is functional. Moreover, expression of dominant-negative or antisense cadherin-6/2 construct in bone marrow-derived mouse stromal ST2 cells, which express only cadherin-6/2, markedly impaired their ability to support osteoclast formation in a mouse coculture model of osteoclastogenesis. Our results suggest that cadherin-6 may be a contributory molecule to the heterotypic interactions between the hemopoietic osteoclast cell lineage and osteoblast/bone marrow stromal cells required for the osteoclast differentiation. Since both osteoclasts and osteoblasts/bone marrow stromal cells are the primary cells controlling physiological bone remodeling, expression of cadherin-6 isoforms in these two cell types of different origin suggests a critical role of these molecules in the relationship of osteoclast precursors and cells of osteoblastic lineage within the bone microenvironment.

Amino Acid Sequence↗
PubMed1998

Smad5 and DPC4 are key molecules in mediating BMP-2-induced osteoblastic differentiation of the pluripotent mesenchymal precursor cell line C2C12.

Since the bone morphogenetic proteins (BMPs) are members of the transforming growth factor-beta (TGF-beta) superfamily that induce the differentiation of mesenchymal precursor cells into the osteogenic cells, we identified the relevant signaling molecules responsible for mediating BMP-2 effects on mesenchymal precursor cells. BMP-2 induces osteoblastic differentiation of the pluripotent mesenchymal cell line C2C12 by increasing alkaline phosphatase activity and osteocalcin production. As recent studies have demonstrated that cytoplasmic Smad proteins are involved in TGF-beta superfamily signaling, we plan to isolate the relevant Smad family members involved in osteoblastic differentiation. We identified human Smad5, which is highly homologous to Smad1. BMP-2 caused serine phosphorylation of Smad5 as well as Smad1. In contrast, TGF-beta failed to cause serine phosphorylation of Smad1 and Smad5. We found Smad5 is directly activated by BMP type Ia or Ib receptors through physical association with these receptors. Following phosphorylation, Smad5 bound to DPC4, another Smad family member, and the complex was translocated to the nucleus. Overexpression of point-mutated Smad5 (G419S) or a C-terminal deletion mutant DPC4 (DPC4 delta C) blocked the induction of alkaline phosphatase activity, osteocalcin production, and Smad5-DPC4 signaling cascades upon BMP-2 treatment in C2C12 cells. These data suggest that activation of Smad5 and subsequent Smad5-DPC4 complex formation are key steps in the BMP signaling pathway, which mediates BMP-2-induced osteoblastic differentiation of the C2C12 mesenchymal cells.

Amino Acid Sequence↗