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Biomedical subjects

E Uchida

Publications and source records attributed to E Uchida.

At least 37 records · Page 2Linked to original sources
PubMed2001

Immunohistochemical detection of apolipoprotein B-100 and immunoglobulins (IgA, IgM, IgG) in the splenic arteries of aging dogs.

Accumulation of lipids and hyalinosis in the splenic arteries of aged dogs are frequently detected by routine histopathologic examinations. The purpose of this study was to pinpoint the localization of canine apolipoprotein B-100 (CApoB-100) and immunoglobulins (IgA, IgM, IgG) in the splenic arteries of aging dogs (n = 80) through the use of immunohistochemical techniques. CApoB-100 deposits were found in the subendothelial space, extracellular matrix, and atheromatous lesions in the tunica media of the arteries in dogs > or = 6 years of age. Foamy cytoplasm of the infiltrated macrophages was also CApoB-100 immunopositive. In dogs > or = 10 years of age, almost all central arteries were CApoB-100 immunopositive. Hyaline deposits within the wall were characterized by immunopositivity against canine IgA, IgM, IgG, and albumin. Lipid accumulation in splenic arteries may be an age-related lesion and a precursor of the atheromatous plaques associated with splenic hemorrhage and infarcts later in life. In addition, deposition of immunoglobulins, probably mediated by immune complexes, may play an important role in the development of canine vascular diseases similar to human disease.

Age Factors↗
PubMed2000

On the mechanism of plasmin-induced platelet aggregation. Implications of the dual role of granule ADP.

Plasmin-induced platelet aggregation has been considered to be a cause of reocclusion after thrombolytic treatment with plasminogen activators. However, little is known regarding the mechanism and regulation of plasmin-induced platelet aggregation. In this study, we demonstrated that plasmin causes the degranulation of platelets, and that ADP released from granules plays a crucial role in the induction of platelet aggregation. This conclusion is supported by results showing that both ADP antagonists and ADPase can inhibit the effect of plasmin on platelets. We also demonstrated that pretreatment of platelets with ADP makes the platelets more sensitive to plasmin, and plasmin-induced platelet aggregation is, therefore, observed at lower concentrations where no aggregation occurs in quiescent platelets. In other words, it is thought that ADP potentiates the plasmin-induced aggregation. The effect of ADP was inhibited by N(6)-[2-(methylthio)-ethyl]-2-(3,3, 3-trifluoropropyl)thio-5'-adenylic acid, monoanhydride with dichloromethylenebisphosphonic acid (AR-C69931), a selective antagonist for the P2T(AC) subtype of P2 receptor, but not by the P2Y1 receptor-selective antagonist adenosine 3'-phosphate 5'-phosphosulfate (A3P5PS). The P2X1 receptor agonist alpha, beta-methylene adenosine 5'-triphosphate (alpha,beta-MeATP) did not mimic the action of ADP. These data indicate that ADP potentiates plasmin-induced platelet aggregation via the P2T(AC) receptor. In addition, epinephrine, a typical G(i) agonist against platelets, could potentiate the plasmin-induced platelet aggregation, suggesting that the signal via the G(i) protein is involved in potentiating the plasmin-induced platelet aggregation, ADP is secreted from platelet granules, and concomitantly works in conjunction with plasmin in a P2T(AC) receptor-mediated manner.

Adenosine Diphosphate↗
PubMed2000

IRES-dependent second gene expression is significantly lower than cap-dependent first gene expression in a bicistronic vector.

The internal ribosome entry site (IRES) has been widely used to coexpress heterologous gene products by a message from a single promoter. However, little is known about the efficiency of IRES-dependent second gene expression in comparison with that of first gene expression. This study was undertaken to characterize the relative expression of IRES-dependent second gene in a bicistronic vector, which was derived from the 5' untranslated regions of the encephalomyocarditis virus (EMCV). IRES-dependent second gene expression was compared with cap-dependent first gene expression in several cultured cell lines and in mouse liver in vivo. The expression of the IRES-dependent second gene ranged from 6 to 100% (in most cases between 20 and 50%) that of the first gene. Second gene expression in a plasmid without the IRES was 0.1-0.8% (with some exceptions) that of the first gene. These findings have important implications for the use of IRES, i.e., care should be taken regarding the decreased capacity of IRES-dependent downstream gene expression as well as in determining which gene should be positioned as the first or second gene in a bicistronic vector.

5' Untranslated Regions↗
PubMed2000

The cytotoxic effects of bile acids in crude bile on human pancreatic cancer cell lines.

Pancreatic cancer frequently causes extrahepatic cholestasis. To identify the direct effects of bile acids in jaundiced serum on pancreatic cancer, the proliferation of PANC-1 and MIA PaCa-2 cells as well as the ultrastructural alteration of PANC-1 cells cultured in crude bile modified media were studied. The growth of these cells in the RPMI-1640 media with or without 1%, 2%, and 4% of the refined crude bile was assessed after 48 and 96 h of incubation. The ultrastructure of PANC-1 cells was investigated by scanning and transmission electron microscopy after 24 and 48 h of incubation. The proliferation of both cell lines in the bile-treated media was greatly inhibited. The inhibitory rates of bile on PANC-1 ranged from 24.1% +/- 3.3% to 66.9% +/- 6.6% (P < 0.01) and those on MIA PaCa-2 ranged from 16.7% +/- 3.8% to 50.7% +/- 5.5%. (P < 0.01). When the bile-added media were replaced, the cells were able to restore their proliferating ability. The PANC-1 cells incubated in the bile-supplied media indicated that the mirovilli, mitochondria, and other organelles had thus been injured. These results suggest that bile acids appear to inhibit the proliferation of PANC-1 and MIA PaCa-2 cells, and the probable inhibitory mechanism is mainly considered to be due to the cytotoxicity of such bile acids.

Bile Acids and Salts↗
PubMed2000

Surface modification of polyethylene balloon catheters for local drug delivery.

Local drug delivery is an attractive approach to the associated problems of percutaneous transluminal coronary angioplasty (PTCA), including arterial injury. The objective of the present research was to deliver a high concentration of a potent anti-thrombin agent, argatroban (ARG), to the vessel wall in order to reduce arterial injury. Local delivery was accomplished by the ionic attachment of drug particles to a modified balloon surface. Surface graft polymerization of ionic monomers to a high-density poly(ethylene) (PE) substrate was performed utilizing ultra-violet (UV) methods. Acrylic acid (AAc) and 2(dimethylamino) ethyl methacrylate (DMAEMA) were successfully grafted onto PE surfaces. Surface grafting was verified by contact angle, X-ray photoelectron spectroscopy, and zeta potential measurements. The amount of ARG adsorbed onto the modified PE surface was highly dependent on the pH of the drug media for both anionic and cationic grafted monomers. The efficacy of local drug delivery to the arterial wall was analyzed using drug-immobilized PE balloon catheters in the rabbit common carotid artery model. High concentrations of ARG (280 nmol/g tissue) were found within the ballooned arterial segment immediately after angioplasty, followed by a decrease after blood flow was restored.

Acrylates↗
PubMed2000

Effect of angiostatin on liver metastasis of pancreatic cancer in hamsters.

The liver is the most common site of metastasis in pancreatic cancer, and there are no promising strategies to treat it. Angiostatin, a kringle-containing fragment of plasminogen, is a potent inhibitor of angiogenesis. The effect of angiostatin on liver metastasis in pancreatic cancer was investigated by using our established hamster model of liver metastasis. Pancreatic cancer cells (PGHAM-1, 1 x 10(6)) derived from N-nitrosobis(2-oxopropyl)amine (BOP)-induced pancreatic tumor in Syrian golden hamsters were transplanted into the spleen of female hamsters, and the animals were subcutaneously injected with angiostatin and saline. Subsequently, the macroscopic appearance of liver surface metastases was evaluated. In addition, histological sections of the liver metastases were analyzed for neovascularization, proliferation, and apoptosis on the basis of von Willebrand factor, argyrophilic nucleolar organizer region (Ag-NOR), and TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining, respectively. The results showed significant tumor growth retardation and inhibition of angiogenesis in metastatic liver tumors in response to treatment with angiostatin. Moreover, the metastases remained in a nearly dormant state due to a balance between apoptosis and proliferation of the tumor, with no detectable side effects. This is the first experimental trial of angiostatin on pancreatic cancer and liver metastasis. The results suggest that angiostatin therapy could be effective against liver metastases of pancreatic cancer.

Angiogenesis Inhibitors↗
PubMed2000

Enzyme-linked immunosorbent assays of canine apolipoproteins B-100 and A-I.

An enzyme-linked immunosorbent assay (ELISA) for canine blood apolipoprotein (apo) B-100 and A-I was developed. The working range for the assay was 1.8 to 28.7 ng/well for apoB-100 and it was 50 to 410 ng/well for apoA-I. The intra- and inter-assay coefficients of variation for the assay for apoB-100 were 5.4 and 6.9%, respectively, and for apoA-I they were 5.8 and 10.6%, respectively. The average concentrations of apoB-100 and A-I in 25 beagles (males, aged 3-4 years) were 0.084 +/- 0.028 (mean +/- SD) mg/ ml and 6.29 +/- 1.55 mg/ml, respectively. The ratios of canine (C) apoB-100 to CapoA-I were 1.41 +/- 0.58%. The respective concentrations in one case of hyperlipidemia with systemic atherosclerosis were 0.454 mg/ml and 11.28 mg/ml (a ratio of 4.03%). These values were larger than those of the controls. These results suggest that the measurements of CapoB-100 and A-I concentrations by this newly developed ELISA are helpful for diagnosis of lipidosis.

Animals↗
PubMed1999

Commitment of neutrophilic differentiation and proliferation of HL-60 cells coincides with expression of transferrin receptor. Effect of granulocyte colony stimulating factor on differentiation and proliferation.

To examine the regulatory mechanisms of proliferation and maturation in neutrophilic lineage cells, we have tried to sort dimethyl sulfoxide (Me(2)SO)-treated HL-60 cells into transferrin receptor (Trf-R) positive (Trf-R(+)) and negative (Trf-R(-)) cells. Differentiated Trf-R(-) cells expressed more formyl-Met-Leu-Phe receptor (fMLP-receptor) and ability of O-(2) genaration, as markers of differentiation, than Trf-R(+) cells, and Trf-R(-) cell differentiation was markedly accelerated by the incubation with granulocyte colony stimulating factor (G-CSF). On the other hand, Trf-R(+) cells had a tendency to proliferate rather than differentiate, and proliferation was enhanced by G-CSF. These results indicate that Trf-R expression coincides with the commitment to proliferate or differentiate of HL-60 cells, and G-CSF accelerates these commitments. G-CSF-induced tyrosine phosphorylation of STAT 3 in Trf-R(-) cells much more than in Trf-R(+) cells. Protein 70 S6 kinase expression was higher in Trf-R(+) cells than in Trf-R(-) cells. Furthermore, p70 S6 kinase was hyperphosphorylated by G-CSF in Trf-R(+) cells, but not in Trf-R(-) cells. Rapamycin, an inhibitor of p70 S6 kinase activity, inhibited G-CSF-dependent proliferation of Trf-R(+) cells and increased fMLP-R expression on these cells. These results suggest that commitment to proliferation and differentiation in Me(2)SO-treated HL-60 cells is preprogrammed and correlated with Trf-R expression, and G-CSF potentiates the cellular commitment. STAT 3 may promote differentiation of Me(2)SO-treated HL-60 cells into neutrophils, while p70 S6 kinase may promote proliferation and negatively regulate neutrophilic differentiation.

Cell Differentiation↗
PubMed1999

The role of STAT3 in granulocyte colony-stimulating factor-induced enhancement of neutrophilic differentiation of Me2SO-treated HL-60 cells. GM-CSF inhibits the nuclear translocation of tyrosine-phosphorylated STAT3.

The role of granulocyte colony-stimulating factor (G-CSF) on neutrophilic differentiation of Me2SO-treated HL-60 cells was studied. G-CSF augmented the functional maturation of Me2SO-treated HL-60 cells in terms of both O-2-generating ability and expression of the formyl-methionyl-leucyl-phenylalanine receptor. G-CSF induced enhancement of cell growth in Me2SO-treated HL-60 cells. These results indicate that G-CSF is a potent enhancer for the differentiation and proliferation of Me2SO-treated HL-60 cells. G-CSF caused the activation of p70 S6 kinase but not mitogen-activated protein (MAP) kinase. On the other hand, G-CSF rapidly induced tyrosine phosphorylation of signal transducers and activators of transcription-3 (STAT3), but did not induce serine727 phosphorylation. From the analysis of confocal laser scanning fluorescence microscopy and differential centrifugation, it was clearly demonstrated that G-CSF induced nuclear translocation of tyrosine-phosphorylated STAT3. The G-CSF-dependent enhancement of neutrophilic differentiation in Me2SO-HL-60 cells was reversely inhibited by granulocyte-macrophage colony-stimulating factor (GM-CSF). Notably, in the presence of GM-CSF, G-CSF induced the tyrosine phosphorylation of STAT3 but failed to induce the nuclear translocation of tyrosine-phosphorylated STAT3. GM-CSF induced activation of not only p70 S6 kinase, but also of MAP kinase. Furthermore, GM-CSF caused the rapid serine727 phosphorylation of STAT3, both in the presence and absence of G-CSF. PD98059, an MEK1 inhibitor, inhibited the G-CSF-dependent serine727 phosphorylation of STAT3 and blocked the inhibitory effect of GM-CSF on G-CSF-dependent nuclear translocation of STAT3. These results suggest that G-CSF-dependent nuclear translocation of STAT3 coordinates with the promotion of neutrophilic differentiation in Me2SO-treated HL-60 cells.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMed1999

Target-cell specificity of fusogenic liposomes: membrane fusion-mediated macromolecule delivery into human blood mononuclear cells.

Fusogenic liposome, a unique vector prepared by fusing ultraviolet-inactivated Sendai virus and liposome, is known to efficiently deliver content into various animal cells through membrane fusion. In this study, we examined the target-cell specificity of fusogenic liposome (FL)-mediated macromolecule delivery into human blood cells using diphtheria toxin fragment A (DTA) as a probe. Among the peripheral blood mononuclear cells (PBMC), FL was able to deliver its encapsulates into CD14+ monocytes and CD4-/CD8- T-cells, but not into CD19+ B-lymphocytes, CD4+ T-cells or CD8+ T-cells. The susceptibility of human leukemia cell lines to FL was similar to that of PBMC; the order of the reactivity was U937 (monoblastic leukemia)>MOLT4, Jurkat (T-lymphoma)>Daudi, BALL1 (B-lymphoma)>K562 (erythroblastic leukemia). Interestingly, FL showed similar binding activity to all of these leukemia cell lines. These findings indicate that, among blood cells, monocytes, monoblastic leukemia cells, CD4-/CD8- T-cells and T-lymphoma cells are preferable targets for FL-mediated macromolecule delivery. This is the first demonstration of the existence of non-permissive cells against FL. Our results also suggest that some molecules on target-cells other than the binding targets of SV-derived protein may participate in fusion between FL and cells.

Diphtheria Toxin↗
PubMed1999

Assessment of transmural coronary blood flow with intraoperative transesophageal color Doppler echocardiography during coronary revascularization.

Intraoperative color Doppler transesophageal echocardiography with a 4- to 7-MHz transducer was performed on 28 consecutive patients who underwent coronary artery bypass grafting to image and evaluate the transmural coronary blood flow before and after cardiopulmonary bypass. The transmural coronary flow was visualized in 26 (92.8%) of 28 patients in the inferior wall and in 13 (46.4%) of 28 patients in the lateral wall. The peak diastolic flow velocity of the transmural coronary artery in the inferior and lateral wall was significantly increased after coronary revascularization in patients with a successful bypass graft to the right coronary artery (from 34.0 +/- 19.7 to 64.9 +/- 30.9 cm/s, P <.001, n = 10) and to the left circumflex coronary artery (from 35.1 +/- 18.6 to 62.1 +/- 21.1 cm/s, P <.001, n = 10). No significant changes were observed in patients with no bypass graft to the right or left circumflex coronary artery. Coronary blood flow can be mapped and the velocity measured with Doppler transesophageal echocardiography with a high-frequency (4- to 7-MHz) transducer. Assessment of the transmural coronary flow may provide valuable information and aid in decision making during surgical revascularization.

Adult↗
PubMed1999

Analysis of K-ras codon 12 point mutations using duodenal lavage fluid for diagnosis of pancreatic carcinoma.

We evaluated the diagnostic significance of the K-ras point mutations at codon 12 in duodenal lavage fluid (DLF) compared with pure pancreatic juice (PPJ). The DLF was easily and safely collected by injecting distilled water into the duodenum and then aspirating through the working channel of the endoscope during endoscopic retrograde cholangiopancreatography. Two types of DLF are collected this way: DLF 1 is collected just after insertion of the endoscope into the duodenum and DLF 2 is collected after cholangiopancreatography and/or collection of the PPJ using secretin. Analysis of K-ras mutations was performed using enriched polymerase chain reaction. In patients with pancreatic carcinoma (PC), K-ras mutations were detected in 14 of 23 (60.9%) in DLF 1, 16 of 21 (76.2%) in DLF 2, 14 of 20 (70.0%) in PPJ, and 19 of 21 (90.5%) in either DLF 1 or DLF 2. In patients with noncancerous pancreatic diseases consisting of pancreatic cystic diseases and chronic pancreatitis, the incidence of K-ras mutations was 2 of 21 (9.5%) in DLF 1 and 7 of 19 (36.8%) in DLF 2. These values were lower than that in PPJ, and there was significant difference between the incidence in DLF 1 and PPJ. These results suggested that DLF may provide a new and useful material for analysis of K-ras codon 12 point mutations in the diagnosis of PC.

Adenocarcinoma↗
PubMed1999

[An evaluation of non-clinical animal data from the point of view of a clinical pharmacologist].

The development of a new chemical entity for human use is a stepwise process based on an assessment of both animal and human data on efficacy and safety of the drug. Clinical pharmacologists always refer to animal data through an Investigator's Brochure (IB) when planning and performing a clinical trial(s). The IB should provide the investigator(s) with useful information to select doses, dosing intervals, and safety monitoring procedures and also to support the clinical management of subjects during the trial(s). Non-clinical animal studies contained in the IB, however, lack a relationship to the pharmacological and toxicological findings of the investigated product(s). Most of the non-clinical animal studies address the methodology and the results obtained, but are lacking in a discussion of the relevance of the findings. The IB should include not only a summary of the findings in each field of animal study but also relationships of the findings through some indicator(s) such as blood and tissue concentrations of the parent drug and/or metabolites. I do hope Pharmaceutical Companies will provide much useful information about their product(s) through the improvement of their system of research and development.

Animals↗
PubMed1999

Two types of peritoneal dissemination of pancreatic cancer cells in a hamster model.

Peritoneal dissemination has an unfavorable impact on the prognosis of pancreatic cancer, and a peritoneal dissemination model was created in hamsters by using an experimental pancreatic cancer to clarify its pathological characteristics. PGHAM-1, a cancer cell line we established from BOP induced pancreatic cancer in Syrian golden hamsters, was inoculated into the abdominal cavity of Syrian golden hamsters. After inoculation, sequential changes in the diaphragm, omentum, and parietal peritoneum, and the metastatic patterns of the PGHAM-1 cells were morphologically investigated by macroscopical, microscopical, and ultrastructural observation. The cancer cells were easily absorbed at the stomata in the diaphragm and milky spots in the omentum, which were absorptive lymphatic structures, and lymphatic metastasis occurred 4 days after inoculation. In the parietal peritoneum, however, the cancer cells attached to and proliferated on the parietal peritoneum where mesothelial cells had exfoliated and the basement membrane was exposed. This process was comparatively time-consuming, and metastasis occurred in the parietal peritoneum at 7 days after inoculation. This study suggested that there might be two patterns of peritoneal dissemination of hamster pancreatic cancer. One route is lymphatic metastasis via stomata in the diaphragm and milky spots in the omentum, and the other is direct metastasis on the parietal peritoneum; each metastasis forms independently.

Animals↗
PubMed1998

Systemic atherosclerosis in dogs: histopathological and immunohistochemical studies of atherosclerotic lesions.

Histopathological and immunohistochemical studies were carried out on five cases of canine systemic atherosclerosis. The five animals were male, and showed hypercholesterolaemia and hypertriglyceridaemia on biochemical analysis of plasma. Histopathologically, atherosclerotic lesions were seen in the aorta and muscular arteries in many organs, including the heart, spleen, kidneys, lungs, pancreas, alimentary tract, urogenital organs, eyes, prostate and urinary bladder. The lesions were characterized by the deposition of lipids and infiltration of lipid-laden foamy cells in the tunica intima and tunica media, sometimes forming fibrofatty plaques, containing abundant sudanophilic material, cholesterol clefts and mineralized material. The lesions started in the tunica intima and extended to the tunica media and tunica adventitia. Immunohistochemical examination with canine apolipoprotein B-100 (CApoB-100) antibody identified the lipids containing low density lipoprotein. Immunoreactivity to CApoB-100 antibody was recognized in the tunica intima, lipid-laden foamy cell cytoplasm and smooth muscle cells in the tunica media, and fibrofatty plaque. These histopathological and immunohistochemical features were similar to those of human atherosclerotic lesions.

Age Factors↗
PubMed1998

Immunohistochemical localization of apolipoprotein B-100 (ApoB-100) and expression of glutathione peroxidase (GSH-PO) in canine atherosclerotic lesions.

We used immunohistochemistry to localize canine Apolipoprotein B-100 (CApoB-100) and glutathione peroxidase (GSH-PO) in canine atherosclerotic lesions. CApoB-100 was deposited in the tunica intima and cytoplasms of infiltrating macrophages in early atherosclerotic lesions. In advanced atherosclerotic lesions, the cystic space of the lesions contained a large amount of CApoB-100 immunoreactive material. Expression of GSH-PO was recognized in the foamy cytoplasm of macrophages and smooth muscle cells in the early and advanced atherosclerotic lesions. These results indicate that expression of GSH-PO is closely associated with the deposition of CApoB-100. In addition, they suggest that, as in human atheromas, low-density lipoprotein (LDL) is peroxidized and changed into modified LDL. Deposition of modified LDL (oxidized or acetylated) may be a critical step in the formation of canine atherosclerotic lesions.

Animals↗
PubMed1998

Clinicopathological studies on association of gallbladder carcinoma and pancreaticobiliary maljunction.

During the past 17 years, 1,722 of 4,832 consecutive patients investigated with endoscopic retrograde cholangiopancreatography (ERCP) were assessed by the radiological criteria of the Japanese Study Group of Pancreaticobiliary Maljunction (PBM). Out of these 1,722 patients, PBM was found in a total of 52, representing 3.0%, of which gallbladder carcinoma was associated with 14. These 14 with gallbladder carcinoma consisted of 10 (62.5%) of the 16 with PBM without association of congenital bile duct dilatation (CBDD) and 4 (11.1%) of the 36 PBM with CBDD. The relationship between PBM and gallbladder carcinoma was closely examined; PBM was noted in 14 (32.6%) of a total of 43 patients with gallbladder carcinoma compared to 38 (2.3%) among the 1,679 patients with various diseases excluding gallbladder carcinoma. Similarly, it was revealed that gallbladder carcinoma was predominantly noted in the 14 (26.5%) of the 52 patients with PBM in contrast to an incidence of 1.7% (29) among the 1,670 patients without PBM. As we studied the characteristic clinical features of the 14 gallbladder carcinoma patients with PBM when compared with 29 of those without PBM, the following was disclosed: on average, the patients with PBM were 10 years younger (49.4 vs 61.4 years in mean age); there was a preponderance of women (0/14 vs 12/17, male-female ratio); there existed a significantly lower incidence of associated gallstone disease (7.1% vs 72.4%). These figures were shown to be statistically significant. We concluded that the results prove a link between the crucial features of gallbladder carcinoma and PBM, and also suggest the promotive role of PBM in carcinogenesis of the gallbladder.

Adenocarcinoma↗