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Biomedical subjects

Y Nagashima

Publications and source records attributed to Y Nagashima.

At least 109 records · Page 6Linked to original sources
PubMed1998

Inhibitory effect of matrilysin antisense oligonucleotides on human colon cancer cell invasion in vitro.

In colorectal cancer, matrilysin (matrix metalloproteinase-7) is mainly produced by the tumor cells themselves and is thought to play an important role in tumor invasion and metastasis. In the study reported here, we examined the effects of matrilysin antisense phosphorothioate oligonucleotides on both the expression of matrilysin and the invasive potential of the human colon cancer cell line CaR-1 in vitro. To select the most specific and potent oligonucleotide sequence, we performed extensive analyses of the binding specificities of all antisense candidates in the GenBank database by using a computer program we developed. As a result, a 15-mer matrilysin-specific antisense oligonucleotide that hybridizes to the coding region of matrilysin mRNA (AS-1) and a random control oligonucleotide (CL-1) were designed. Reverse transcription-polymerase chain reaction and western blot analysis demonstrated that 10 microM AS-1 suppressed matrilysin expression at both the mRNA level (92%) and protein level (64%). In vitro invasion assays demonstrated that this same concentration of AS-1 inhibited the ability of cells to invade a reconstituted basement membrane by 50% as compared with the ability of untreated cells to do so. On the other hand, CL-1, which had the same length and GC content as AS-1, did not show any inhibitory effect. These results demonstrate that the antisense oligonucleotide AS-1 inhibits matrilysin activities in a sequence-specific manner and suggest that AS-1 has the potential to be used as an anti-metastatic agent in an in vivo experimental model of colon cancer.

Antineoplastic Agents↗
PubMed1998

[A case of pulmonary arteriovenous fistula beneath the surface of the lung].

A 19-year-old woman complaining of exertional dyspnea was admitted to our hospital with an abnormal shadow on the left side of the chest. Laboratory examination revealed polycythemia and hypoxemia. Pulmonary angiogram demonstrated a pulmonary arteriovenous fistula beneath the surface of the left S10. Partial resection of the left S10 was performed. The wall of the arteriovenous fistula was flimsy and seemed to rupture easily. The sister of this patient also had a peripheral pulmonary arteriovenous fistula and suffered from repeating epistaxis. Rendu-Osler-Weber disease was diagnosed in both, and the sister underwent partial resection of the right S7, which contained the fistula. Their postoperative courses were uneventful. Hemothorax and hemoptysis are lethal complications of arteriovenous fistulae. In order to avoid the rupture of fistulae, surgical resection is the most reliable treatment. Pulmonary arteriovenous fistulae beneath the surface of the lung should be resected.

Adolescent↗
PubMed1998

[Japan Coma Scale as a grading scale of subarachnoid hemorrhage: a way to determine the scale].

BACKGROUND: The grading scale for subarachnoid hemorrhage (SAH) with inter-grade outcome differences is essential for evaluating the effectiveness of newly developed therapeutic modalities. Although Hunt's grade and WFNS scale have been widely used, these grading scales do not meet this requirement. We previously proposed a revised WFNS scale based solely on the Glasgow Coma Scale (GCS) that has intergrade outcome differences of high-level significance. The Japan Coma Scale (JCS) has been long and widely used in Japan. The purpose of this study is to show whether it is possible to determine a reasonable SAH grading scale based on the JCS and to show a way to determine an SAH grading scale. PATIENTS AND METHODS: We retrospectively analyzed 1398 consecutive cases of aneurysmal SAH operated on within Day 7 of the latest onset. The preoperative JCS and GCS were evaluated just before the surgery and the Glasgow Outcome Scale (GOS), analyzed with numerical transformation (1 = dead to 5 = good recovery), was estimated at 6 months after the onset. All 510 possible combinations of scores of JCS were statistically tested under the following 2 assumptions; (1) JCS = 0 and JCS = 100 fall into a single independent grade. (2) No other single JCS score should fall into a single grade. RESULTS: The outcome differences between JCS 0 and 1, and 100 and 200 are significant. The outcome difference between JCS 30 and 100 is relatively higher than any other set of 2 scores of JCS. Only 5 combinations are practical among the candidates to be analyzed. Out of 510 combinations, the following combination shows the highest inter-grade outcome differences; I (JCS = 0, n = 375, mean GOS = 4.78) II (JCS = 1, 2; n = 310; mean GOS = 4.47) III (JCS = 3-30; n = 476; mean GOS = 3.96) IV (JCS = 100; n = 96; mean GOS = 3.10) V (JCS = 200, 300; n = 141; mean GOS = 2.33). In JCS, the mean outcome of JCS = 3 is worse than those of JCS = 10, 20, and 30. The outcome difference between JCS 0 and 1 is only significant in patients over 60 years old. CONCLUSION: Taking all the 510 possible combinations of JCS into consideration, we obtained a reasonable combination containing 5 grades. Although this grading scale showed good inter-grade outcome differences, JCS is not preferable to GCS as a consciousness evaluation system in the acute phase of SAH. We emphasize the importance of this way to determine a grading scale with a combinatorial approach, which can be applicable for re-evaluating the grading scales in the future.

Age Factors↗
PubMed1998

Determination of Na channel blockers in paralytic shellfish toxins and pufferfish toxins with a tissue biosensor.

The biosensor consisted of a sodium electrode and covered with the frog bladder membrane within a flow cell was tested for the estimation of tetrodotoxin (TTX) and saxitoxin (STX). This sensor was applied to detect very low amounts of the Na+ channel blockers, STX and TTX, in different shellfishes and swellfishes. A good agreement was obtained between TTX activities determined by mouse assay and amounts of Na+ channel blockers estimated by frog membrane sensor. The lowest level of TTX (fg) that can be determined by frog membrane sensor does not cause human poisoning. The channel blockers in short-necked clam, which was assumed to be STX, were monitored by this sensor continuously every week for one year. It was discovered that the STX content increased from July until September and then decreased from October until March. The biosensor proposed here may be used for the estimation of STX and TTX conventionally in the future.

Animals↗
PubMed1998

[Two cases of solitary peripheral pulmonary arterial aneurysm].

We report two very rare cases of solitary peripheral pulmonary arterial aneurysm. Case 1: An 83-year-old man treated for myocardial infarction died of multiple organ failure and hemoptysis. Autopsy disclosed rupture of a pulmonary arterial aneurysm 1.5 cm in diameter in the right A3. Case 2: A 75-year-old man was found to have a pulmonary arterial aneurysm, 3 cm in diameter of the right A1. Surgical treatment was not indicated, because of severe pulmonary emphysema. The aneurysms in these two cases were idiopathic: (1) there were no histologic findings that would implicate specific chronic inflammation. (2) the period from the occurrence of secondary pulmonary hypertension to hemoptysis was short, and there was no (3) history of trauma.

Aged↗
PubMed1998

Elevated expression of membrane-type 1 and 3 matrix metalloproteinases in rat vascular smooth muscle cells activated by arterial injury.

Matrix metalloproteinases (MMPs) play critical roles in tissue remodeling under various physiologic and pathologic conditions. We recently reported the expression of three membrane-type MMPs (MT-MMPs) by cultured vascular smooth muscle cells (SMCs) of rats (Shofuda et al, 1997). To investigate the roles of the MT-MMPs in the matrix remodeling of blood vessels, expression of MT1-MMP and MT3-MMP was examined in normal and balloon-injured rat carotid arteries by in situ hybridization and immunohistochemistry. Both MT-MMP mRNAs were detected in the intimal-dedifferentiated SMCs, but were negligible in the medial SMCs or in any of normal vascular cells. To elucidate the regulatory mechanism for the MT-MMPs expression, effects of various factors on cultured rat SMCs were also examined. MT1-MMP mRNA was constantly expressed at a high level, and its expression was weakly increased by treatment with interleukin-1beta or tumor necrosis factor-alpha. When the cells were incubated with type IV collagen, the MT1 -MMP expression was markedly decreased. On the other hand, expression of MT3-MMP mRNA was strongly increased by platelet-derived growth factor and fibronectin. These results suggest that type IV collagen may act as a negative regulator for the expression of MT1-MMP in the medial SMCs, whereas platelet-derived growth factor and fibronectin may up-regulate MT3-MMP expression under pathologic conditions. Furthermore, the elevated expression of MT1-MMP and MT3-MMP in SMCs was well associated with their dedifferentiated phenotype.

Animals↗
PubMed1997

Regulation and immunohistochemical analysis of stress protein heme oxygenase-1 in rat kidney with myoglobinuric acute renal failure.

Intramuscular injection of hypertonic glycerol solution to rats results in acute renal injury. In this model, the proximal tubules are characteristically damaged. After glycerol injection renal glutathione (GSH) levels drastically decreased. On the other hand, stress protein heme oxygenase-1 (HO-1) was induced. When N-acetyl cysteine was administered to rats before 1 h glycerol injection, renal function was obviously improved. In this condition, the renal GSH content were sustained in the normal levels and HO-1 protein levels were decreased compared with those of glycerol-treated rats. Induction of HO-1 was accompanied by reduced renal GSH content. In addition, to investigate whether the location of HO-1 protein induced by glycerol injection is restricted to injured region or not in the kidney, we determined the localization of HO-1 protein using immunohistochemical staining. HO-1 protein was identified in the epithelia of the distal tubules, Henle's loop and collecting ducts, but not in the injured proximal tubules.

Acetylcysteine↗
PubMed1997

PACAP-27 causes negative and positive dromotropic effects in anesthetized dogs.

While pituitary adenylate cyclase-activating polypeptide (PACAP) has been identified radioimmunologically in the rat heart, the physiological role of PACAP has not been elucidated in the regulation of the atrioventricular conduction in the heart. We, therefore, determined the dromotropic effects of PACAP-27 injected into the cannulated atrioventricular node artery in the autonomically decentralized heart of the open-chest, anesthetized dog. PACAP-27 caused transient positive followed by negative dromotropic responses in a dose-dependent manner, whereas vasoactive intestinal peptide (VIP) caused only a positive dromotropic response. Atropine and tetrodotoxin blocked the negative dromotropic response to PACAP-27 and after blockade PACAP-27 caused only a positive dromotropic response. Tetrodotoxin and propranolol did not affect the positive dromotropic response to PACAP-27 in atropine-treated dogs. PACAP-27 altered the atrio-His bundle interval but did not alter the His-ventricle interval. These results demonstrate that PACAP-27 prolongs the atrio-His bundle interval due to the liberation of acetylcholine from parasympathetic nerves and decreases it by a non-adrenergic mechanism in the dog heart in situ.

Animals↗
PubMed1997

Uterine carcinosarcoma is derived from a single stem cell: an in vitro study.

Expression of intermediate filaments (IFs) has been suggested to be a reliable marker for differentiating epithelial and non-epithelial tumors. Moreover, the c-erbB-2 and p53 genes are considered to be involved relatively early in the process of human carcinogenesis. In order to elucidate the origin of uterine carcinosarcomas, we analyzed IF, c-erbB-2 and p53 expression in and the ultrastructural characteristics of clones derived from a human uterine-carcinosarcoma cell line, EMTOKA. The expression of IFs and other proteins in the EMTOKA clones was identical to that in the EMTOKA cell line. It and its 7 clones all expressed cytokeratins 8, 17, 18 and 19, vimentin, epithelial membrane antigen, S-100, myoglobin, type-II collagen, alpha-smooth-muscle actin, placental alkaline phosphatase and epidermal-growth-factor receptor. The c-erbB-2 and p53 expression levels of all the cell types of the EMTOKA cell line and its clones were the same. Interestingly, an ultrastructural study showed that the EMTOKA cell line and its clones at early and late passages possessed the characteristics of epithelial cell types without either transitional forms between the epithelial and stromal components or differentiation into sarcomatous components. The results of this study lend particular support to the combination tumor hypothesis that a precursor (stem) cell gives rise both to epithelial and to mesenchymal components during the histogenesis of uterine carcinosarcoma, the epithelial component of which appears to be dominant, suggesting that the established cell lines derived from a common stem cell.

Biomarkers, Tumor↗
PubMed1997

Expression of matrilysin in vascular endothelial cells adjacent to matrilysin-producing tumors.

Matrilysin is believed to play important roles in tumor progression and metastasis. In the present study, we analyzed matrilysin-producing cells in various human cancer tissues by immunohistochemistry and in situ hybridization. Tumor cells in colorectal carcinomas, pancreatic carcinomas, transitional-cell carcinomas of the kidney and small-cell lung carcinomas were frequently positive for matrilysin. In addition, we found that endothelial cells of arterioles and venules adjacent to matrilysin-positive tumors expressed matrilysin mRNA and protein. The endothelial cells adjacent to matrilysin-negative tumors and those in normal tissues were negative for matrilysin. Furthermore, analyses by casein zymography, Western blotting and RT-PCR showed that matrilysin was weakly expressed by cultured human umbilical vein endothelial cells. Our results suggest that the expression of matrilysin in vascular endothelial cells and in tumor cells may be regulated by common soluble factors, and that endothelial cell-derived matrilysin may contribute to tumor angiogenesis and tumor metastasis.

Animals↗
PubMed1997

Expression of trypsin in vascular endothelial cells.

Proteinases produced by vascular endothelial cells are expected to play important roles in many biological processes. Here we report that human vascular endothelial cells express trypsinogen-2 mRNA and its protein product in culture. The trypsinogen production was stimulated by a tumor promoter and associated with cell growth. In situ hybridization analysis showed that the trypsinogen gene was significantly expressed in vascular endothelial cells around gastric tumors and in patients with disseminated intravascular coagulation (DIC). These results suggest the possible roles of endothelial cell-derived trypsin in tumor angiogenesis and abnormal blood coagulation.

Cells, Cultured↗
PubMed1997

Novel polypeptide toxins with crab lethality from the sea anemone Anemonia erythraea.

The sea anemone Anemonia erythraea was found to contain polypeptide toxins with crab lethality as well as hemolysins. Three polypeptide toxins (AETX I, II and III) were isolated by gel filtration on Sephadex G-50 and reverse-phase HPLC on TSKgel ODS-120T. A geographic variation in toxin composition was suggested. The LD50 against crabs of AETX I, II and III were estimated to be 2.2, 0.53 and 0.28 microg/kg, respectively, but none of the toxins showed lethality in mice. The amino acid sequences of the three toxins were deduced from sequencings of the whole molecules and their enzymatic fragments. Amino acid analyses and molecular mass determinations supported the accuracy of the deduced sequences. AETX I, comprising 47 amino acid residues including 6 half-Cys residues, is an analog of sea anemone type I toxins. On the other hand, AETX II and III, which are highly homologous with each other, are quite distinct from the known sea anemone polypeptide toxins in that they are composed of 59 residues including 10 half-Cys residues. Interestingly, both toxins have sequence similarities with neurotoxins isolated from the Brazilian 'armed' spider Phoneutria nigriventer.

Amino Acid Sequence↗
PubMed1997

Halcurin, a polypeptide toxin from the sea anemone Halcurias sp., with a structural resemblance to type 1 and 2 toxins.

The aqueous extract of the sea anemone Halcurias sp. belonging to the suborder Endocoelantheae was found to be potently lethal to crabs, although it showed neither lethal activity in mice nor hemolytic activity. A polypeptide toxin (named halcurin) with a LD50 of 5.8 micrograms/kg against crabs was isolated by gel filtration on Sephadex G-50 and reverse-phase high-performance liquid chromatography on TSKgel ODS-120T. The complete amino acid sequence of halcurin comprising 47 residues was elucidated by sequence analysis of the native molecule and its enzymatic fragment. Comparison with the known sea anemone polypeptide toxins (types 1-3), which are all from members of the suborder Nynantheae, revealed a high sequence homology (49-74%) of halcurin with type 2 toxins. Also, halcurin has several residues conserved for only type 1 toxins. These results, together with the fact the Halcurias sp. is a more primitive species than members of Nynantheae, suggest that type 1 and 2 toxins have evolved from a common ancestor with a sequence more similar to halcurin.

Amino Acid Sequence↗
PubMed1997

Expression of gelatinase A, tissue inhibitor of metalloproteinases-2, matrilysin, and trypsin(ogen) in lung neoplasms: an immunohistochemical study.

Lung cancer is a heterogeneous tumor in terms of clinical and biological behavior, and its aggressiveness depends on its invasive and metastatic properties. Matrix metalloproteinases and serine proteinases are believed to play a crucial role in invasion and metastasis of malignant tumor cells. In the present study, the authors evaluated immunohistochemically the expression of gelatinase A; tissue inhibitor of metalloproteinases-2 (TIMP-2), an inhibitor of gelatinase A; matrilysin; and trypsin(ogen) in 67 lung tumors from a variety of histological types including 17 squamous cell carcinomas, 16 adenocarcinomas, 15 small cell carcinomas, and 12 carcinoids. Interestingly, normal bronchial, bronchiolar, and alveolar epithelial cells expressed gelatinase A, TIMP-2, matrilysin, and trypsin(ogen) at varying frequencies and intensities. Bronchial smooth muscle cells and cartilage cells expressed gelatinase A alone, whereas endothelial cells, fibroblasts, and macrophages expressed gelatinase A and TIMP-2. Gelatinase A was expressed at high levels in most lung tumors examined (47% to 80%). TIMP-2 was also expressed at high levels except in the small cell carcinomas, which showed TIMP-2 expression at a lower frequency (60%) compared with other types of lung tumors (80% to 100%). Although matrilysin was expressed by tumor cells of all the histological types at various frequencies (13% to 63%), its expression was most common in adenocarcinomas. Expression of trypsin(ogen) was observed almost exclusively in adenocarcinomas (56%); other types of lung tumors expressed trypsin(ogen) far less frequently (0% to 12%). The present results, taken together with those of previous studies, suggest that gelatinase A is associated with malignant behavior of all the types of lung tumors, whereas its activity may be controlled by the endogenous inhibitor TIMP-2. The aggressive clinical behavior of small cell carcinoma may be attributable, at least in part, to a loss of the inhibitory effect of TIMP-2, as a significant proportion of these tumors showed negative or low levels of TIMP-2 expression. Matrilysin and trypsin(ogen) expressions are unlikely to be correlated with the aggressiveness of lung tumors. The expression of trypsin (ogen) may rather reflect the differentiation of adenocarcinoma cells toward normal airway epithelial cells.

Adult↗
PubMed1997

Isolation and amino acid sequences of two Kunitz-type protease inhibitors from the sea anemone Anthopleura aff. xanthogrammica.

Two protease inhibitors (AXPI-I and -II) were isolated from the sea anemone Anthopleura aff. xanthogrammica by a combination of acetone precipitation, gel filtration on Sephadex G-75, cation-exchange fast protein liquid chromatography (FPLC) on Mono S and reverse-phase HPLC on TSKgel ODS-120T. Both inhibitors are basic polypeptides, and their amino acid compositions are characterized by the presence of six half-Cys residues and the absence of Met and Trp. They are potently active against trypsin; inhibition of other serine proteases (alpha-chymotrypsin and elastase) is also displayed by only AXPI-I. However, the inhibitors show no affinity for metallo-proteases and cysteine proteases. Analyses of the N-terminal portion and enzymatic fragments established their complete amino acid sequences comprising 58 residues. The overall sequence homology and the conserved location of all half-Cys residues confirmed that the A. aff. xanthogrammica inhibitors belong to the Kunitz-type family.

Amino Acid Sequence↗
PubMed1997

Adult ganglioneuroblastoma of the anterior mediastinum.

A case of ganglioneuroblastoma occurring in the anterior mediastinum of a 79 year-old man is reported. The tumor was mainly composed of neuroblasts with occasional ganglion cells. Foci of melanin-laden cells were also identified. Immunohistochemistry revealed that the tumor cells showed both schwannian and melanocytic differentiation with immunoreactivity to anti-S100 protein and anti-HMB45 antibodies. In addition, the tumor contained several microcysts lined by squamous epithelial and one lymphoid tissue abundant in T lymphocytes, which appeared to be derived from thymic tissue. This case is unique in that neuroblastoma group tumors including ganglioneuroblastoma is uncommon in the elderly and in the thymic region, and rarely shows melanocytic differentiation. To the best of our knowledge, this case is a tumor of neuroblastoma group occurring in the eldest patient.

Aged↗
PubMed1997

Effects of PACAP-38 on the SA nodal pacemaker activity in autonomically decentralized hearts of anesthetized dogs.

Pituitary adenylate cyclase-activating polypeptide (PACAP) receptors exist, but the physiologic role of PACAP is unclear in the heart in situ. We investigated effects of PACAP-38 on sinus rate and on the negative chronotropic response to acetylcholine (ACh) or stimulation of the intracardiac parasympathetic nerve fibers to the sinoatrial nodal region in the automatically decentralized heart of the open chest, anesthetized dog. PACAP-38 (0.1-1 nmol) injected directly into the sinus node artery caused transient positive followed by negative chronotropic responses. Both pretreatment with atropine and tetrodotoxin inhibited the negative chronotropic responses to PACAP-38. However, hexamethonium did not block the negative responses to PACAP-38. After treatment with PACAP-38 (0.1-1 nmol), ACh induced atrial fibrillation significantly (p < 0.01). On the other hand, the negative chronotropic responses to intracardiac parasympathetic stimulation were not changed. These results suggest that (a) PACAP-38 induces negative chronotropic responses and liberates ACh from intracardiac postganglionic parasympathetic nerves, and that (b) PACAP-38 reduces ACh-induced atrial fibrillation threshold in the dog heart in situ.

Acetylcholine↗