Biomedical subjects

H Shimada

Publications and source records attributed to H Shimada.

At least 289 records · Page 16Linked to original sources

PubMed1998

Matrilysin-specific antisense oligonucleotide inhibits liver metastasis of human colon cancer cells in a nude mouse model.

Human colon cancer frequently develops liver metastasis. Matrilysin (MMP-7), the smallest member of the matrix metalloproteinase (MMP) family, is commonly produced by human colon carcinoma cells and has been suggested to be involved in the progression and metastasis of this type of cancer. In the present study, we tested the effect of a matrilysin-specific antisense phosphorothioate oligonucleotide on liver metastasis of the human colon carcinoma cell line WiDr in nude mice. In culture, the antisense oligonucleotide moderately inhibited the secretion of matrilysin by WiDr cells. Injection of WiDr cells into the spleen of nude mice produced many metastatic tumor nodules in the liver. When the antisense oligonucleotide was injected daily into the mice for 11 days, the formation of the metastatic tumor nodules was strongly inhibited in a dose-dependent manner. An inhibition of liver metastasis of over 70% was obtained at a dose of 120 micrograms of the oligonucleotide per mouse. The antisense oligonucleotide did not inhibit tumor growth in spleen and in liver. A scrambled control oligonucleotide had no effect on liver metastasis of WiDr cells. Our results demonstrate an important role of matrilysin in liver metastasis of human colon cancer and the therapeutic potential of matrilysin antisense oligonucleotides for the prevention of metastasis.

Aged↗

PubMed1998

Cadmium suppresses apoptosis induced by chromium.

Cadmium and chromium are both well-known human carcinogens, and common exposures to these metals are not infrequent. Recent studies have shown that hexavalent chromium induces apoptosis in Chinese hamster ovary (CHO) cells, suggesting an association of apoptosis with carcinogenesis. In contrast, induction of apoptosis by cadmium has been inconsistently observed. The present study was designed to determine if cadmium could induce apoptosis in CHO cells and if common exposure to cadmium and chromium would modify any apoptotic response. Apoptosis was evaluated by both agarose gel and in situ end-labeling methods. Apoptosis was observed at 48 h after treatment with 300 microM chromium (Na2CrO4) for 2 h. Cadmium alone at concentrations of 1, 5, or 10 microM (as CdCl2) did not induce apoptosis in these cells even at times up to 72 h after treatment. However, when CHO cells were concurrently exposed to cadmium and chromium, chromium-induced apoptosis was markedly suppressed in a cadmium concentration-related fashion. Cadmium did not consistently modify the cytotoxic effects of chromium, and significant increases in metallothionein were not induced by these metal treatments. These findings indicate that cadmium can block chromium-induced apoptosis. The suppression of apoptosis by cadmium may be a significant aspect of its carcinogenic mechanism.

Animals↗

PubMed1998

Talipexole protects dopaminergic neurons from methamphetamine toxicity in C57BL/6N mouse.

The effect of protection of dopaminergic neurons by talipexole, a dopamine (DA) agonist, is investigated on a methamphetamine (MA)-induced parkinsonism model of mice (C57BL/6N). The reduction of tyrosine hydroxylase activity in the striatum 72 h after MA (5 mg/kg every 2 h, four times) treatment was attenuated by the administration of talipexole (0.25 mg/kg or 1.0 mg/kg) prior to the administration of MA. In an in vitro experiment, talipexole inhibited the adduction reaction of hydroxyl radicals to salicylate. Taken together, these data suggest that the protective effect of talipexole on DA neurons is, in part, caused by the hydroxyl radical-scavenging action of the drug.

Animals↗

PubMed1998

Matrix metalloproteinases-2 and -9 are expressed in human neuroblastoma: contribution of stromal cells to their production and correlation with metastasis.

Neuroblastoma, the second most common solid childhood tumor, can be a highly invasive and metastatic form of cancer. To assess the role of matrix-degrading proteases in this cancer, we have examined the expression of matrix metalloproteinases (MMPs) and their corresponding tissue inhibitors of metalloproteinases (TIMPs) in 7 human neuroblastoma cell lines and 24 primary untreated tumors. MMP-2 (gelatinase A) and MMP-9 (gelatinase B) were the only two MMPs expressed. MMP-2 was detected predominantly in an inactive proform in all tumor cell lines and tumor tissue extracts. The lack of MMP-2 activation in cell lines was attributed to the absence of expression of a membrane-type MMP (MT1-MMP), which activates proMMP-2, and to the abundant expression of TIMPs, particularly TIMP-2. Immunohistochemical analysis of tumor tissue samples indicated that MMP-2 was present in both tumor cells and stromal cells. In contrast, MMP-9 was not expressed by neuroblastoma cell lines but was present in inactive and active forms in extracts from tumor tissues. Immunohistochemical analysis of positive specimens indicated that MMP-9 was predominantly present in stromal, vascular, and perivascular cells surrounding nests of tumor cells. There was no correlation between the levels of these MMPs and the MYCN copy number or the histopathological phenotype. However, there were higher levels of MMP-2 and MMP-9 in stage IV (metastatic) disease when compared with stages I and II (noninvasive and nonmetastatic) or IV-S (P < 0.05).

Collagenases↗

PubMed1998

Mitochondrial NADH-quinone oxidoreductase of the outer membrane is responsible for paraquat cytotoxicity in rat livers.

We investigated the existence of an NADH-dependent paraquat (PQ) reduction system in rat liver mitochondria (Mt) in respect to the cytotoxic mechanisms of PQ. The outer membrane fractions, free from the contamination of inner membranes but with a few microsomes, catalyzed rotenone-insensitive NADH, but not NADPH, oxidation by menadione or PQ. Anti-NADH-cytochrome b5 reductase antibody and its inhibitor p-hydroxymercuribenzonate did not inhibit the NADH-PQ reduction activity. Therefore, the respiratory systems of the inner membranes and microsomal cytochrome P450 systems could not have been responsible for the reaction. Dicoumarol, an inhibitor of NAD(P)H-quinone oxidoreductase (NQO), dose dependently suppressed the NADH oxidation in the outer membrane via PQ as well as menadione, with I50 values of 190 (for menadione) and 150 microM (for PQ). Because of a lower sensitivity to NADPH and the higher doses of dicoumarol required for its inhibition, the activity in the outer membrane may be an "NADH-quinone oxidoreductase" which partly differs from the NQO previously reported. This outer membrane enzyme produced superoxide anions in the presence of both NADH and PQ and was too tightly membrane-bound to be extracted by Triton X-100 and deoxycholate. From these results, we concluded that the free radical-producing mitochondrial NADH-quinone oxidoreductase is a novel oxidation-reduction system participating in PQ toxicity. This is in good agreement with our previous results showing that PQ selectively damaged Mt in vivo and in vitro, resulting in cell death (K.-I. Hirai et al., 1992, Toxicology 72, 1-16).

Animals↗

PubMed1998

Enzymic confirmation of reactions involved in routes to astaxanthin formation, elucidated using a direct substrate in vitro assay.

An in vitro assay procedure for the carotenoid (beta-ionone ring) 3,3'-hydroxylase and 4,4'-oxygenase has been developed that enables efficient conversion of non-radiolabeled carotenoid substrates added directly into aqueous solution. The following enzymic conversions were demonstrated and apparent kinetic constants (Vmax, Km, and specificity constants) obtained: (a) 3,3'-hydroxylase (from Agrobacterium aurantiacum and Alcaligenes sp. strain PC-1) converted phoenicoxanthin (adonirubin) to astaxanthin, 3-hydroxyechinenone to 4-ketozeaxanthin (adonixanthin), 3'-hydroxyechinenone to 4-ketozeaxanthin, as well as echinenone to 4-ketozeaxanthin via 3- and 3'-hydroxyechinenone; (b) 4,4'-Oxygenase (from A. aurantiacum, Alcaligenes sp. strain PC-1 and Haematococcus pluvialis) converted 4-ketozeaxanthin to astaxanthin, 3-hydroxyechinenone to phoenicoxanthin, 3'-hydroxyechinenone to phoenicoxanthin, and echinenone to canthaxanthin. Determination of substrate specifities allowed assessment of biosynthetic routes to astaxanthin formation and demonstrated that pathways via mono-hydroxylated and ketolated products are enzymically feasible.

Alcaligenes↗

PubMed1998

Arylsulfatase exists as non-enzymatic cell surface protein in sea urchin embryos.

The physiological role of arylsulfatase (Ars) and its function during development have yet to be satisfactorily defined in any species, though the proteins are widely distributed and the genes have been cloned from various organisms. Here we report the dual location of two types of Ars in sea urchin embryos. The majority of sea urchin Ars does not exhibit enzyme activity and is extracellularly distributed in aboral ectoderm cells (nonenzymatic Ars). Only a small portion has enzyme activity and is localized in lysosomal vesicles (enzymatic Ars). The elution pattern of Ars proteins processed by DEAE-cellulose or analytical gel-column chromatography reveals that although the molecular radius of enzymatic Ars differs from that of nonenzymatic Ars, they have the same charge. Furthermore, sedimentation analysis shows that purified Ars of sea urchin embryos is soluble in the absence of divalent cations but becomes insoluble in the presence of Ca2+ or Mg2+. Taken together, the present results suggest that non-enzymatic Ars is a new member of the cell surface component or extracellular matrix. It is possible that this cell surface Ars plays an important role in morphogenesis of sea urchin embryos.

Animals↗

PubMed1998

Membrane conformations and their relation to cytotoxicity of asimicin and its analogues.

Certain plant species belonging to the family Annonaceae produce Annonaceous acetogenins, which are a unique class of long-chain fatty acid derivatives with potent cytotoxicity. Putative protein targets of the acetogenins are membrane-associated proteins, including complex I. Asimicin and its analogues constitute a class of Annonaceous acetogenins containing two tetrahydrofuran (THF) rings with hydrocarbon chains tethered to each ring; an alpha,beta-unsaturated gamma-lactone ring is terminal to one of the alkyl chains. The compounds examined in this study differ in the length of the alkyl chain between the THF rings and the lactone ring. The positions of both the THF and the lactone rings within liposomal membranes were determined by proton (1H) nuclear magnetic resonance spectroscopy. The depth of membrane penetration of acetogenins, coupled to membrane diffusion, controls the conformation of acetogenins as they diffuse to an active site. Based on 1H intermolecular nuclear Overhauser effects (NOEs), the THF rings of all acetogenins studied reside near the polar interfacial head group region of the DMPC. This was corroborated by 1H two-dimensional NOE spectroscopy and differential scanning calorimetry studies. The 1H difference NOE spectra indicated that the lactone rings of asimicin and parviflorin, the latter of which has two fewer carbons in its alkyl chain, are located below the glycerol backbone in the membrane. In contrast with asimicin and parviflorin, the lactone ring of longimicin B, an asimicin analogue with an alkyl chain four carbons shorter, resides close to the midplane in the membrane. This was corroborated by manganese-induced broadening studies. Since the THF rings are located near the center of the acetogenin molecules and the lactone ring is terminal to a long alkyl chain, these observations indicate that an asimicin-type acetogenin can be in either sickle-shaped or U-shaped conformations, depending on the length of the alkyl chain between the THF rings and the lactone ring. Interestingly, longimicin B does not exhibit significant cytotoxicity, but parviflorin is as cytotoxic as asimicin. The cytotoxicity of the asimicin-type of acetogenins would seem to be strongly related to the membrane conformation. This is the first report elucidating the conformation of Annonaceous acetogenins in membranes.

Calorimetry, Differential Scanning↗

PubMed1998

Structure and function of a sea urchin orthodenticle-related gene (HpOtx).

Two distinct types of orthodenticle-related proteins (HpOtxE/L) have been implicated as transcription activators of the aboral ectoderm-specific arylsulfatase (Ars) gene. Here, we describe the structure of HpOtx gene and present evidence that mRNAs of HpOtxE/L are transcribed from a single HpOtx gene by altering the transcription start site and by alternative splicing. By transactivation experiments, we have also demonstrated that HpOtxL activates the Ars promoter in the gastrula-stage embryo.

Animals↗

PubMed1998

Bone marrow and liver mutagenesis in lacZ transgenic mice treated with hexavalent chromium.

The mutagenic effects of the hexavalent chromium compound K2CrO4 in lacZ transgenic mice (Muta Mouse) were investigated at two sampling times. K2CrO4 was administered intraperitoneally to five male mice per treatment group at a single dose of 40 mg/kg. The animals were sacrificed on days 1 and 7 after the treatment. Mutant frequencies in the bone marrow and liver were analyzed by the positive selection method using Escherichia coli C (galE-) strain and phenyl beta-D-galactoside. K2CrO4 induced a significant increase in mutant frequency in the bone marrow on day 1, but not on day 7 after the treatment. In the liver, on the other hand, a significant induction in the mutant frequency was seen on day 7, whereas no induction was observed on day 1. The reason for the different responses to the mutagenic activity of K2CrO4 between these organs may be related to their cell turnover rates. The mutations induced by K2CrO4 in the bone marrow may have occurred in more differentiated cells than stem cells, and the rapid proliferative activity may have caused a rapid decrease in mutated cells by day 7. These results suggest that experiments on mutagenesis should be done with more than one sampling point, a short expression time in addition to a longer one, so as to detect mutations induced in organ with high cell proliferation.

Animals↗

PubMed1998

Assessment of the proliferative activity of superficial esophageal carcinoma using MIB-1 immunostaining for the Ki-67 antigen.

BACKGROUND AND OBJECTIVES: Lymph node metastasis or vascular invasion may occur in superficial esophageal squamous cell carcinoma when it invades to or into the muscularis mucosae. Therefore, the correlation between histopathological characteristics and the proliferative activity of superficial esophageal carcinoma was investigated. METHODS: Thirty-eight cases of esophageal squamous cell carcinoma, including 14 cases of mucosal carcinoma and 24 cases of submucosal carcinoma, who underwent surgical resection without preoperative treatment, were studied using monoclonal antibody MIB-1 for the Ki-67 antigen immunohistochemically. The labeling index (LI) was calculated with a computed image analyzer. RESULTS: The LI of MIB-1 at the invasive tip of m3 carcinoma was significantly higher than that of m1 or m2 carcinoma (P < 0.01). The LI at the invasive tip was significantly higher than that at the core of sm2 (P < 0.05) and submucosal carcinoma overall (P < 0.01). The LI values at both the invasive tip and core of poorly differentiated carcinoma in submucosal carcinoma were higher than that of well or moderately differentiated carcinoma with a significant difference (P < 0.05). The LI at the invasive tip of submucosal carcinoma with lymph node metastasis or lymphatic invasion was significantly higher than that without them (P < 0.05). CONCLUSION: Proliferative activities of cancer cells in superficial esophageal squamous cell carcinoma, immunostaining with the MIB-1, were related to the depth of invasion, differentiation, lymph node metastasis, and lymphatic invasion with a significant difference.

Antibodies, Monoclonal↗

PubMed1998

The localisations in liposomal membranes of the tetrahydrofuran ring moieties of the annonaceous acetogenins, annonacin and sylvaticin, as determined by 1H NMR spectroscopy.

The positions of the tetrahydrofuran (THF) ring moieties of annonacin (a mono-THF ring Annonaceous acetogenin) and sylvaticin (a non-adjacent bis-THF ring Annonaceous acetogenin) within liposomal membranes made of dimyristoylphosphatidylcholine (DMPC) were determined by proton (1H) nuclear magnetic resonance (NMR) spectroscopy. Based on 1H intermolecular nuclear Overhauser effects (NOEs), the THF rings of both acetogenins studied reside near the polar interfacial head group region of the DMPC. Recently, we have reported that the THF rings of a series of asimicin type of Annonaceous acetogenins (with adjacent bis-THF rings) also reside near the interfacial head group of DMPC. We can now conclude that the Annonaceous acetogenins, containing either mono-, adjacent bis-, or non-adjacent bis-THF ring moieties, have their THF ring moieties at the interfacial region of membranes, i.e., the THF ring moiety seems to serve a role as an anchor in the lipid membranes. This may be related to the uniquely potent bioactivities that Annonaceous acetogenins exhibit at their enzyme-inhibitory sites within mitochondrial and plasma membranes.

Antineoplastic Agents, Phytogenic↗

PubMed1998

Evaluation of a new staging system by the Japanese Research Society for Gastric Cancer.

The feasibility of the new classification of stage grouping by the Japanese Research Society for Gastric Cancer was evaluated. During the 22-year period between January 1975 and December 1996, a total of 1294 patients with primary gastric cancer underwent laparotomy at the Department of Surgery, Chiba University; 1222 had their lesions removed during the gastrectomy and 72 remained nonresected. Cases of direct operative death totaled 17 (1.3%). Five hundred patients (38.6%) died of a relapse of the original cancer and 42 (3.2%) died of other diseases within the followup period. Six patients (0.5%) were lost during the followup. The 5-year cumulative patient survival rates of the seven stages of the new stage grouping were distinctly proportional, and the differences were also statistically significant except between stages IIIb and IVa. The two major revised points in the new stage grouping, new classification of the depth of cancer invasion, and new stage grouping by a mosaic combination of the degree of invasion and lymph node metastasis were thus found to be reasonable based on the actuarial 5-year survival rates of the subgroups in the same stage. The present study also showed that the classification of stage IV still requires further discussion.

Evaluation Studies as Topic↗

PubMed1998

Micrometastatic colorectal cancer lesions in the liver.

A surgical resection of metastatic liver lesions from colorectal cancer contributes to an improved prognosis. However, the postoperative recurrence rate remains high, particularly in the residual liver. This is probably the result of the failure to detect small lesions. In the present study, we histologically examined the presence of intrahepatic micrometastases, which are considered to be related to recurrence in the residual liver. Intrahepatic micrometastases were histologically examined in 31 resected specimens of 25 patients undergoing a hepatic resection because of metastasis to the liver from colorectal cancer. Micrometastases were found in 14 of 25 cases (56.0%). They were located in the portal veins, central veins, sinusoid, and bile ducts. The longest distance from the main metastasis was 38.2 (mean 7.5 +/- 8.0) mm. The size of the macrometastases became larger, and the frequency of micrometastases and the distance of micrometastases from macrometastases had a tendency to increase. Continuous invasion of the macrometastases into the micrometastases through the vasculature or bile duct was also observed. These results suggested that some micrometastases observed in the metastatic liver from colorectal cancer were thus seeded from the primary lesions, while other micrometastases originated from the macrometastatic lesions as satellite lesions.

Colorectal Neoplasms↗

PubMed1998

Rate pressure product and oxygen saturation in tourists at approximately 3000 m above sea level.

OBJECTIVES: Using modern transportation technology, many travelers easily access moderate altitudes of approximately 3000 m above sea level. In the present study the effects of this altitude on cardiovascular parameters were studied among office workers dwelling at sea level. METHODS: Heart rate, blood pressure, arterial oxygen saturation (SpO2), and electrocardiography were monitored before and after Master's double-step exercise at 2700 and 3700 m. The test consisted of stepping onto and off of two 23-cm steps for 3 min at a predefined rate. RESULTS: The resting values recorded for the heart rate and mean blood pressure at 2700 and 3700 m did not statistically significantly differ from those noted at sea level. However, the increases in these values after exercise were significantly greater at high altitude. The rate pressure product more than doubled after exercise at 3700 m. Electrocardiographic abnormalities were observed in some cases. The postexercise blood lactate concentration was significantly higher at 3700 m than at sea level or at 2700 m, suggesting that the oxygen supply-demand relationship was not balanced at this altitude. Furthermore, exercise provoked an acute reduction in SpO2 at 2700 and 3700 m but showed no effect at sea level. CONCLUSION: These observations suggest that the oxygenation status of the heart might be at risk in many travelers and workers during and after exercise load at an altitude of approximately 3000 m.

Adult↗

PubMed1998

New bioactive triterpenoids from Melia volkensii.

Bioactivity-directed fractionation of the root bark of Melia volkensii resulted in the isolation of two new natural products, meliavolkinin (1) and melianin C (3), together with two known compounds, 1,3-diacetylvilasinin (2) and melianin B (4). Jones oxidation of 4 gave compounds 3, 23,24-diketomelianin B (5), and 16,23,24-triketomelianin B (6). The structures of the new compounds were elucidated by spectral and chemical data. Compounds 1-6 all showed marginal cytotoxicities against certain human tumor cell lines, while 5 showed selective cytotoxicities for the human prostate (PC-3) and pancreatic (PACA-2) cell lines with potencies comparable to those of adriamycin.

Antineoplastic Agents, Phytogenic↗

PubMed1998

Detection of regional lymph node metastases in colon cancer by using RT-PCR for matrix metalloproteinase 7, matrilysin.

Lymph node metastasis is the most important prognostic factor in colon cancer. However, more accurate screening for metastasis than that afforded by conventional pathology remains elusive. We have employed a reverse transcriptase-polymerase chain reaction (RT-PCR) assay for a matrix metalloproteinase (MMP), 'matrilysin', because this gene is epithelial-specific and consistently expressed in colorectal cancer cells. The sensitivity of this assay was examined with the matrilysin-producing rectal cancer cell line 'CaR-1'. Matrilysin mRNA was detected in this system when more than 10(4) matrilysin-positive cells existed in a lymph node of ordinary size. Fourteen of 15 (93%) primary colon cancers and none of the surrounding normal tissues expressed matrilysin. All 10 histologically-positive lymph nodes were positive for matrilysin, while of 60 histologically-negative lymph nodes, eight were positive for matrilysin. When the additional sequential sectioning and histological re-examination was performed on five of these eight 'matrilysin-positive, but histologically-negative' lymph nodes, micrometastases were detected in three. Only one of the lymph nodes that were histologically-positive, but negative by matrilysin assay was from a patient with colon cancer in which matrilysin was not detected. In conclusion, RT-PCR assay for matrilysin is a sensitive method for detecting occult metastases in patients with colon cancer, and may complement histologic examination.

Adult↗

PubMed1998

Proximal cis-regulatory elements of sea urchin arylsulfatase gene.

Expression of the arylsulfatase (HpArs) gene in the sea urchin, Hemicentrotus pulcherrimus, is regulated in spatially, as well as temporally, during development. To address the cis-regulatory elements involved in this regulation, we performed reporter assays using variously deleted or mutated promoter and regulatory elements of the HpArs gene, accompanied by gel mobility shift assay and foot printing. Results show that two regions, PU1 (-72 b.p. to -56 b.p.), which is similar to SpZ12-1 and/or Oct-1 motif, and the PD1 site (+133 b.p. to +142 b.p.), which is homologous to the binding sites of Rel family transcription factors and/or AGIE-BP1, are related to the regulation of expression of the HpArs gene. Furthermore, an HpArs enhancer element called C15, which is located 3 kb.p. downstream from the transcription start site, activates the HpArs promoter. We also report that the enhancer activity of the C15 fragment was mediated by elements, PU1 and PD1.

Animals↗