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Biomedical subjects

K Uchida

Publications and source records attributed to K Uchida.

At least 757 records · Page 42Linked to original sources
PubMed1995

Antiproliferation and colony-forming inhibition activities of recombinant feline interferon (rFeIFN) on various cells in vitro.

The antiproliferative and colony inhibiting activities of recombinant feline interferon (rFeIFN Type I) against various cells in vitro were examined. Feline and canine cells were both sensitive to rFeIFN. To inhibit the growth of feline cells by 50% approximately 5 x 10(2) to 1 x 10(3) U/mL rFeIFN was required and maximum activity was achieved at a concentration of 1 x 10(5) U/mL. Approximately 5 x 10(3) to 5 x 10(4) U/mL rFeIFN was necessary to inhibit the growth of canine cells by 50%. The antiproliferative and colony inhibiting activities of rFeIFN on canine cells appeared to be cell-specific and dose-dependent. However, human, monkey and hamster cells were resistant to rFeIFN. We suggest that rFeIFN might be useful for treatment of feline and some canine neoplastic conditions.

Animals↗
PubMed1995

Expression of intercellular adhesion molecule-1 on cultured glomerular endothelial cells by pro-inflammatory cytokines and lipopolysaccharide.

Infiltration of leukocytes and mononuclear cells into the glomeruli is an early pathological finding in human and experimental glomerulonephritis. However, the cellular and molecular basis for cell infiltration into the glomeruli is not yet completely understood. In addition, there is little information on the expression of intercellular adhesion molecule-1 (ICAM-1) on glomerular cells. In the present study, we investigated the expression of ICAM-1 on cultured bovine glomerular endothelial cells (GEN) and its regulation by the pro-inflammatory cytokines, interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1) and lipopolysaccharide (LPS). Immunocytochemical staining showed that ICAM-1 molecules were constitutively expressed on the surface of GEN. In flow cytometric and ELISA analyses, ICAM-1 molecule expression on GEN was significantly upregulated by IL-1 beta, MCP-1 and LPS in a dose-dependent manner, but not by IL-6. LPS was the most potent inducer of ICAM-1 molecule expression on GEN. The effects of IL-1 beta, MCP-1 and LPS were observed as early as 4 h and reached a maximal level by 18 h. These results suggest that ICAM-1 on GEN can participate in the infiltration of mononuclear cells into glomeruli in human and experimental glomerulonephritis.

Animals↗
PubMed1995

Granulation in amine-storage organelles of mouse megakaryocytes: X-ray microprobe analysis and radioautography.

The mechanisms and the processes of the storage of bivalent cations, ATP and 5-hydroxytryptamine (5HT) in the precursors of the amine-storage organelles of megakaryocytes were studied at the electron microscopic level. Although the precursors of the amine-storage organelles in the megakaryocytes fixed with glutaraldehyde and osmium tetroxide were empty, the electron opaque granules were observed in these organelles of the freeze-substituted megakaryocytes cut onto ethylene glycol. X-ray microprobe analysis demonstrated that they contained P, Mg and Ca. Quantitative differences in bivalent cations in the granules were not observed between megakaryocytes and blood platelets. Electron opaque uranaffin-reaction products were observed in the precursors of the amine-storage organelles of the megakaryocytes after treatment with the uranaffin reaction for ATP. However, few chromaffin positive granules were observed in the precursors of the amine-storage organelles after the chromaffin reaction for monoamines. Radioautographic analysis demonstrated that blood platelets avidly took up 3H-5HT but megakaryocytes were not able to accumulate 3H-5HT in vivo. These results indicate that megakaryocytes do not yet acquire the well developed uptake system of 5HT in vivo, while they readily accumulate cations and ATP in the precursors of the amine-storage organelles.

Adenosine Triphosphate↗
PubMed1995

The forms and the levels of fecal PMN-elastase in patients with colorectal diseases.

OBJECTIVES: To compare the form of polymorphonuclear leukocyte (PMN)-elastase in feces with that in plasma and to investigate the usefulness of measuring fecal PMN-elastase levels in patients with colorectal diseases. METHODS: We examined PMN-elastase complexed with alpha 1-antitrypsin (alpha 1-AT), chymotrypsin, and alpha 2-macroglobulin by ELISA in feces and plasma. Fecal levels of total PMN-elastase were determined in patients with colonic polyp (N = 19), colonic cancer (N = 20), ulcerative colitis (UC; N = 36), colonic Crohn's disease (CD; N = 26), and in control subjects (N = 20). RESULTS: Most PMN-elastase was not complexed with alpha 1-AT, chymotrypsin, or alpha 2-macroglobulin in feces, whereas most plasma PMN-elastase was complexed with alpha 1-AT. Fecal concentrations and daily fecal excretion of PMN-elastase were significantly increased in patients with active UC (medians 54.8 micrograms/g, 15.14 mg/day) and active CD (41.5 micrograms/g, 10.24 mg/day) compared to those values in control subjects (0.6 micrograms/g, 0.11 mg/day) and in patients with colonic cancer (2.5 micrograms/g, 0.33 mg/day). In inactive UC and CD, these values (3.4 micrograms/g, 0.52 mg/day and 5.2 micrograms/g, 0.59 mg/day, respectively) were significantly lower than in active UC and CD, respectively. In UC, all patients whose rectal biopsies showed infiltration of PMN had high fecal PMN-elastase levels. CONCLUSIONS: Our results suggest that the measurement of fecal PMN-elastase concentrations are useful for monitoring the disease activity of UC and CD, especially when evaluating whether intestinal inflammation has disappeared completely.

Adult↗
PubMed1995

[Correlation between pulmonary vascular changes and hemodynamic parameters during exercise before and after mitral valve surgery].

Sixteen cases with mitral valvular disease were studied with regard to the correlation between pathological changes in the pulmonary vasculature and pulmonary hemodynamics during exercise before and after surgery. In muscular pulmonary arteries obtained by open lung biopsy, medial wall thickness (MWT) was 13.8 +/- 3.2% and intimal thickness (IT) was 37.3 +/- 13.5%. MWT was correlated with IT (r = 0.60, p < 0.05). Wall thickness in pulmonary veins was 8.21 +/- 1.8%. Preoperative mean pulmonary arterial pressure (MPAP) during exercise was 45.9 +/- 9.4 mmHg, and decreased significantly to 38.1 +/- 11.3 mmHg postoperatively. However, in 4 patients, MPAP during exercise increased after surgery. Pulmonary vascular resistance (PVR) during exercise was unchanged before and after surgery (2.74 +/- 1.90 U.M2-->2.69 +/- 1.3 U.M2). MWT was significantly correlated with preoperative MPAP at rest only. In patients showing an increase of MPAP during exercise postoperatively, mean MWT was 16.2% and mean IT was 47.4%. In conclusion our data suggest that postoperative pulmonary hemodynamics does not improve in patients whose MWT exceeds 15% and IT exceeds 40% and that irreversibility may be due to pulmonary vascular lesions.

Adult↗
PubMed1995

Histopathology and pathophysiology of secondary hyperparathyroidism due to chronic renal failure.

Between 1973 and 1992, 300 patients underwent parathyroidectomy for secondary hyperparathyroidism due to chronic renal failure in our departments. Using parathyroid glands obtained at operation, histopathological studies were performed, and to estimate pathophysiology DNA analysis of parathyroid cell nuclei and calcium-regulated parathyroid hormone (PTH) secretion in vitro were estimated. PTH mRNA expression was evaluated by in situ hybridization. The typical histopathological findings were asymmetric enlargement, nodularities and increased number of oxyphil cells. Secondary hyperplasia was divided into 2 types: diffuse and nodular type hyperplasia. In the histopathological study nodular hyperplasia indicated more aggressive proliferation. In DNA analysis the relative number of scattered cells in the DNA synthesis phase was significantly greater in nodular than in diffuse hyperplasia. The half of the maximal inhibition of PTH secretion for calcium (the set-point) in the cells from nodular hyperplasia was higher than in the cells obtained from diffuse hyperplasia. However, there was no difference in expression of PTH mRNA in nodular and diffuse hyperplasia. These data suggested that nodular hyperplasia was more progressively hyperplastic, had more aggressive proliferative activities and showed more abnormal regulation of PTH secretion. These results imply that to prevent graft-dependent recurrent hyperparathyroidism after parathyroidectomy, the nodular hyperplastic tissue should not be autografted.

Calcium↗
PubMed1994

Molecular analysis of T cell receptor V beta chain to detect leukemia cell clonality in patients by adaptor ligation-mediated polymerase chain reaction.

We have developed a simple and rapid method to analyze the clonality of leukemia cells. After three rounds of amplification by adaptor-ligation polymerase chain reaction (PCR), the cDNA is cut with AluI, HaeIII, RsaI, and Sau3AI, and analyzed by polyacrylamide gel electrophoresis. The size of the restriction fragments is compared to that of the published restriction fragments size each TCR-beta subfamily V region. The sensitivity of adaptor-ligation PCR restriction enzyme analysis (AL-PCR-REA) was 10(-4) MOLT-4 T-ALL cell population in the normal peripheral blood lymphocytes (PBL). Application of AL-PCR-REA to PBL and bone marrow (BM) cells from eight clinical leukemia samples indicated that a detection sensitivity was rather low, but revealed the clonality of all eight clinical samples. This AL-PCR-REA method can detect clonality without the need for either radioisotopes or sequencing procedures.

Base Sequence↗
PubMed1994

Michael addition-type 4-hydroxy-2-nonenal adducts in modified low-density lipoproteins: markers for atherosclerosis.

It has been proposed that plasma low-density lipoprotein (LDL) undergoes oxidative modification before it can give rise to foam cells in atherosclerosis. Oxidation of LDL generates a variety of reactive aldehyde products including 4-hydroxy-2-nonenal (HNE), which may covalently attach to the LDL apolipoproteins. We here present direct evidence that HNE derivatization of LDL forms Michael addition-type adducts of HNE with histidine and lysine residues of apolipoprotein B-100 (apoB) and also demonstrate the utility of an antibody specific to the HNE adducts generated in the LDL treated with HNE or oxidatively modified by Cu2+ or cultured endothelial cells. HNE adducts present in the LDL that had been treated with HNE were attested to be Michael addition-type adducts on the basis of the fact that incubation of LDL with 1 mM HNE (2 h, 37 degrees C) resulted primarily in the formation of Michael addition-type HNE-histidine (39.9 mol/mol of LDL) and HNE-lysine (19.3 mol/mol of LDL) adducts. An enzyme-linked immunosorbent assay (ELISA) and an SDS-polyacrylamide gel electrophoresis (SDS-PAGE)/immunoblot analysis of HNE-modified LDL demonstrated that these HNE adducts were detectable with the HNE-specific antibody affinity-purified with the Michael adduct (HNE-histidine) as a ligand. The following lines of evidence indicated the presence of Michael addition-type HNE adducts in the oxidatively modified LDL in vitro: (i) Amino acid analysis of LDL that had been treated with Cu2+ (24 h, 37 degrees C) demonstrated the presence of a Michael addition-type HNE-histidine adduct (7-9 mol/mol of LDL).(ABSTRACT TRUNCATED AT 250 WORDS)

Aldehydes↗
PubMed1994

Formation of 8-hydroxy-2'-deoxyguanosine and 4-hydroxy-2-nonenal-modified proteins in human renal-cell carcinoma.

To study the possible involvement of reactive oxygen species (ROS) in the tumor biology of human renal-cell carcinoma (RCC), we analyzed 35 cases of RCC for 2 parameters of oxidative damage: 8-hydroxy-2'-deoxyguanosine (8-OHdG), a mutation-prone DNA-base-modified product, was measured by means of high-performance liquid chromatography (HPLC) with an electrochemical (EC) detector, and 4-hydroxy-2-nonenal (HNE)-modified proteins were measured with a polyclonal antibody against HNE-modified proteins. A 54% higher content of 8-OHdG was found in RCC than in the corresponding non-tumorous kidney, suggesting that the DNA of RCC is more exposed to ROS than is the DNA of non-tumorous kidneys. Immunohistochemistry for HNE-modified proteins showed a distinct staining pattern of fine to coarse granularity in the cytoplasm of RCC (n = 15), implying that lipid peroxidation products are located in cytoplasmic organelles. These results suggest that RCC constitutionally elaborates more ROS than is produced by the non-tumorous parts of kidneys. No correlation was found between clinical stage, histology, age or sex and the 2 parameters examined.

8-Hydroxy-2'-Deoxyguanosine↗