Verotoxin-producing Escherichia coli O157:H7 carried by the housefly in Japan.
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Adult↗
Biomedical subjects
A Matsuda
Publications and source records attributed to A Matsuda.
Effects of D-methionine-containing solution on tumor cell growth in vitro.
The effects of a nutrition therapy with D-methionine (Met)-containing solution were investigated in cell cultures of the AH109A cell line. The growth of AH109A hepatoma cells in culture media with D-Met-supplemented medium, L-Met-supplemented medium (control) and Met-free medium was compared. The D-Met-supplemented medium inhibited the cell growth to an extent similar to that manifested in the Met-free medium. The total free amino acid concentrations in the control medium decreased by approximately 40% on day 6 post-culture. However, the free amino acid concentrations in D-Met-supplemented and Met-free media did not change. Furthermore, alanine, which was not added to RPMI-1640, was detected in the control medium on day 6 post-culture. These results suggest the possibility of application of D-Met-containing solution to cancer patients receiving total parenteral nutrition.
Amino Acids↗
Identification and immunohistochemical localization of annexin II in rat cornea.
PURPOSE: We have identified annexin II mRNA expression in the rat cornea and demonstrated immunolocalization of annexin II in normal and injured corneas. Furthermore, to investigate possible interaction between annexin II and its extracellular ligand tenascin during corneal wound healing, we also examined tenascin expression simultaneously. METHODS: Total RNA was extracted from the corneal tissue of male Wistar rats as well as from the cell cultures of corneal epithelial cells and keratocytes. cDNA was obtained by reverse transcription (RT). Annexin II mRNA expression was examined by polymerase chain reaction (PCR). Western blot analysis of annexin II protein was performed with protein samples that were obtained from the corneal tissue and cell cultures. In addition, the localization of annexin II and that of tenascin were clarified by immunohistochemical analysis, using both uninjured and epithelial scraped corneas. RESULTS: RT-PCR analysis revealed that annexin II mRNA was expressed in corneal tissues, epithelial cells and keratocytes. Western blot analysis of corneal epithelium and keratocytes showed a 38kDa band that corresponded to the molecular weight of annexin II. Immunohistochemical study showed that annexin II was present in keratocytes as well as the basal cells of corneal epithelium in the central cornea, and basal/suprabasal cells of the limbal epithelium. Positive annexin II immunoreactivity is translocated from cytoplasm to the cell periphery/extracellular position during the epithelial wound healing process. Annexin II and tenascin are coexpressed on the basal surface of limbal epithelium and the leading edge of the healing epithelium. The reappearance of cytoplasmic annexin II staining at the periphery of the cornea correlated with the epithelial cell proliferation. CONCLUSIONS: Annexin II is abundantly expressed in corneal tissue. The translocalization of annexin II protein suggests its role in the corneal epithelial migration during wound healing. The colocalization of annexin II and tenascin in migrating and proliferating corneal epithelial cells suggests that annexin II-tenascin interaction may play a role in epithelial wound healing.
Animals↗
Synthetic study on carbocyclic analogs of cyclic ADP-ribose, a novel second messenger: an efficient synthesis of cyclic IDP-carbocyclic-ribose.
An efficient synthesis of cyclic IDP-carbocyclic-ribose, as a stable mimic for cyclic ADP-ribose, was achieved. 8-Bromo-N1-carbocyclic-ribosylinosine derivative 10, prepared from N1-(2,4-dinitrophenyl)inosine derivative 5 and an optically active carbocyclic amine 6, was converted to 8-bromo-N1-carbocyclic-ribosylinosine bisphosphate derivative 15. Treatment of 15 with I2 in the presence of molecular sieves in pyridine gave the desired cyclic product 16 quantitatively, which was deprotected and reductively debrominated to give the target cyclic IDP-carbocyclic-ribose (3).
Adenosine Diphosphate Ribose↗
N6-methoxyadenine in damaged DNA has two faces in property of Watson-Crick base pairing.
In order to investigate mutation mechanism with oxyamine, two DNA dodecamers containing 2'-deoxy-N6-methoxyadenosine have been synthesized and their crystal structures have been determined. The dodecamers are associated in B form duplexes. The methoxy groups attached to the adenine bases do not affect the DNA conformation significantly. Electron densities clearly show that N6-methoxyadenine moiety forms Watson-Crick type pairing with both, thymine and cytosine bases. Such two faces in pairing are the origin of pyrimidine transition mutation.
Adenine↗
Synthesis and structure-activity relationships of novel nikkomycin analogs: inhibitors of the fungal cell wall biosynthesis enzyme chitin synthase.
A series of novel nikkomycin analogs, which inhibited chitin synthase, the fungal cell wall biosynthesis enzyme, has been synthesized and evaluated their inhibitory activities.
Aminoglycosides↗
Cytotoxic mechanism of 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd).
The molecular mechanism of cell death induced by 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd: Figure 1), a potent inhibitor of RNA synthesis, was performed using mouse mammary tumor FM3A cells and human fibrosarcoma HT1080 cells. ECyd induced the characteristics of apoptosis on these cells, such as morphological changes, DNA fragmentations (Figure 2), and caspase-3-like protease activation. General caspases inhibitor (Z-Asp-CH2-DCB) inhibited these changes and cell death. We also found that ECyd induced DNA and 28S ribosomal RNA (rRNA) fragmentations. Though the mechanisms of rRNA fragmentations haven't revealed, it suggests that translational function of the treated cells should be disturbed. These results indicate that antitumor mechanism of ECyd are characteristics of apoptosis on the cells and rRNA fragmentations is one of the death events resulted inhibition of RNA synthesis.
Animals↗
The first synthesis of herbicidin B, a tricyclic-sugar adenine nucleoside antibiotic, using samarium diiodide-promoted aldol-type C-glycosidation reaction as a key-step.
A first total synthesis of the nucleoside antibiotic herbicidin B (1) was achieved in which a novel aldol-type C-glycosidation reaction promoted by samarium diiodide (SmI2) was used as a key step. Construction of the desired stereochemistry of the tricyclic-sugar moiety was successfully achieved by conformational restriction strategy based on repulsion between adjacent bulky protecting groups on the pyranose ring.
Anti-Bacterial Agents↗
X-ray analysis of a DNA dodecamer containing 2'-deoxy-N4-methoxycytidine.
The crystal structure of DNA dodecamer with the sequence of d(CGCAAATTXGCG), where X is 2'-deoxy-N4-methoxycytidine, has been determined by X-ray analysis. The dodecamers form a double helix with B-form conformation. The electron density indicates that the two modified cytosine bases respectively make a pair with the adenine bases on the opposite strand in a manner of Watson-Crick geometry and that the methoxy groups are in anti conformation to the N3 atom.
Base Pairing↗
Antisense oligodeoxynucleotides: useful tool for search and assessment of new targets for anti-malarial drugs.
We investigated about targeting for new antimalarial drugs using antisense (AS) oligodeoxynucleotides (ODNs). Synthetic nuclease-resistant ODNs (phosphorothioate (PS) ODNs and ODNs containing 4'alpha-C-(2-aminoethyl)thymidines (4'-amino ODNs)) which target mitochondrial succinate dehydrogenase (SDH) iron-sulfur subunit (IP), had antimalarial activity (EC50; about 1.0 microM). Furthermore we showed that intra-parasitic SDH IP mRNA levels, which were detected using quantitative RT-PCR assay, were decreased 13% of control after the 24 h expose to SDH IP AS. From the results, we conclude that SDH has potential as the target for novel antimalarials, and AS ODNs is effective for search and assessment of targets for new antimalarial drugs.
Animals↗
Crystallization and preliminary analysis of a DNA dodecamer of d(CGCGmo6AATCCGCG) containing 2'-deoxy-N6-methoxyadenosine: change in crystal packing with different humidity.
The DNA dodecamer of (CGCGmo6AATCCGCG) containing 2'-deoxy-N6-methoxyadenosine has been crystallized for X-ray analysis in order to investigate the effects of the modified adenosine on base pairing. It has been found that the crystal changes from one form to another during data collection in a manner similar to a phase transition. The two crystal structures show that this phenomenon, ascribed to differences in humidity, is correlated with a change in the contact angle between the two duplexes.
Crystallization↗
Isolation of deoxycytidine kinase from Ehrlich carcinoma cells by affinity chromatography based on a substrate analog, 2'-C-cyano-2'-deoxy-1-beta-D-arabinofuranosyl-N4-palmitoylcytosine++ +.
Deoxycytidine kinase from Ehrlich carcinoma cells was purified 10400-fold by ammonium sulfate fractionation and affinity chromatography using Sepharose 4B coupled to 2'-C-cyano-2'-deoxy-1-beta-D-arabinofuranosyl-N4-palmitoylcytosine , with a yield of 45%. The purified enzyme preparation showed a single major band with a molecular weight of 32000 on SDS-PAGE. The enzyme phosphorylated deoxyadenosine, deoxyguanosine, cytidine, and several deoxycytidine analogues as well as deoxycytidine. Also, the kinetic parameters of the enzyme for the substrates were estimated.
Animals↗
Nucleosides and nucleotides. 186. Synthesis and biological activities of pyrimidine carbocyclic nucleosides with a hydroxyamino group instead of a hydroxymethyl group at the 4'-position of the sugar moiety.
Pyrimidine carbocyclic nucleosides with a hydroxyamino group instead of a hydroxymethyl group at the 4'-position of the sugar moiety were designed as potential antitumor and/or antiviral agents. Pd (O)-catalyzed reactions of enantiomerically pure (+)-(1R,4S)-4-[(tert-butyldiphenylsilyl)oxy]-1-(ethoxycarbonylo xy)-2- cyclopentene (9) with N3-benzoylthymine and -uracil gave carbocyclic nucleosides 10 and 11. Subsequent Pd (O)-catalyzed reactions of N3-benzoyl-1-[(1R,4S)-4-(ethoxycarbonyloxy)-2-cyclopenten-1- yl]thymine (14) and -uracil (15) with O-benzylhydroxylamine smoothly gave the hydroxyamino-substituted carbocyclic nucleosides 16 and 17. From these nucleosides, the target compounds were prepared after deprotection or further reactions. The 2',3'-didehydro-2',3'-dideoxythymidine (D4T) analogue 20 was the most effective compound, with IC50 values of 27.3 and 34.5 microM against KB and L1210 cells in vitro. Carbocyclic analogues of uridine and cytidine (29 and 32) were less effective than 20 against both cell lines.
Animals↗
Synthesis of conformationally restricted analogs of baclofen, a potent GABAB receptor agonist, by the introduction of a cyclopropane ring.
Conformationally restricted analogs of baclofen (2), i.e., 5, 6, and their enantiomers ent-5, and ent-6, the conformations of which were restricted by introducing a cyclopropane ring, were designed as potential GABAB receptor ligands. Reaction of (R)-epichlorohydrin [(R)-7] and (4-chlorophenyl)acetonitrile in the presence of NaNH2 in benzene/tetrahydrofuran gave chiral cyclopropane derivatives 11 and 12, which were then converted into the target compounds 5 and 6, respectively. Their corresponding enantiomers, ent-5 and ent-6, were also synthesized starting from (S)-epichlorohydrin [(S)-7].
Animals↗
Germline MEN1 mutations in sixteen Japanese families with multiple endocrine neoplasia type 1 (MEN1).
OBJECTIVE: Multiple endocrine neoplasia type 1 (MEN1) is a syndrome of endocrine tumors involving the parathyroids, anterior pituitary and enteropancreatic neuroendocrine tissues, and is inherited in an autosomal dominant manner. Recently, the gene responsible for this syndrome, MEN1, was positionally cloned in 11q13. We aimed to assess the significance of MEN1 gene diagnostics in families with MEN1. DESIGN: Sixteen probands of familial MEN1 and their 40 family members were subjected to the study. METHODS: Full-length sequencing of the open reading frame and exon-intron boundaries in the MEN1 gene was performed with probands of familial MEN1. Family members were examined for the identified mutation in the proband. RESULTS: We identified heterozygous germline mutations of the MEN1 gene in all of 16 Japanese MEN1 families examined, achieving the highest detectability of MEN1 mutations in familial MEN1 among studies that examined more than 10 families. Eleven kinds of the identified MEN1 germline mutations were novel. More than half were nonsense or frameshift mutations resulting in a premature stop codon (9/15; 60%), and no mutation hot spots or no apparent genotype-phenotype relationships were observed, in support of the results of other studies. We identified 40 mutant MEN1 gene carriers and 16 non-carriers in the course of the present study in those families. CONCLUSIONS: Analysis of the germline mutations in the MEN1 gene, providing significantly useful clinical information to probands and family members of MEN1, should be considered as a standard procedure and categorized as belonging to Group 1 cancer predisposition testing by the American Society of Clinical Oncology.
Adult↗
[Rapid-acting insulin analogues].
Two rapid-acting insulin analogues, Lys(B28), Pro(B29)-human insulin (insulin lispro) and Asp(B28)-human insulin (insulin aspart) are developed and introduced into the clinical trials or applications recently. The replacement of the natural amino acid sequence at the position 28 of the B-chain of human insulin results in an insulin molecule with reduced self association. Proline at position B28 near the COOH terminal of the B chain of human insulin is important for the dimerization of insulin molecules. These two insulin analogues exist in solution as a monomeric form and display faster pharmacodynamic action than human regular insulin. In healthy volunteers when injected subcutaneously, serum insulin concentration peaks more than twice higher and in less than half the time, compared to regular insulin. The glucose infusion rate to maintain the euglycemia level peaked in about half the time, but the total glucose infused during the euglycemic clamp is not significantly different from that of regular insulin. These rapid absorbed insulin analogues significantly inhibited the postprandial plasma glucose excursion more effectively even injected just before each meal in type 1 diabetic patients. The shorter duration of insulin action reduced the risk of severe hypoglycemia in nighttime or late postprandial time. The long-term treatment with this analogues improved HbA1c levels. Use of these insulin analogues as a preprandial bolus injection improves the quality of life of insulin-treated patients.
Diabetes Mellitus, Type 1↗
[Atypical chronic myeloid leukemia presenting with trilineage dysplasia and IgG (lambda) type monoclonal gammopathy].
A 78-year-old man was diagnosed as leukocytosis in February 1994. Physical examination revealed marked hepatosplenomegaly. A peripheral blood examination disclosed 95,090/microliter leukocytes without hiatus leukemicus, 6.5 g/dl Hb, and 15.0 x 10(4)/microliter platelets. The neutrophil alkaline phosphatase score was 27, and serum VB12 was above 1,600pg/ml. IgG was identified as monoclonal immunoglobulin of type lambda. Bone marrow specimens demonstrated marked granulocytic hyperplasia. Neither the Philadelphia chromosome (Ph1) nor BCR gene rearrangement was detected; hence, the diagnosis of Ph1 (-) chronic myeloid leukemia (CML) was made. The patient was treated with hydroxyurea and low-dose VP-16 with no improvement, and died of pneumonia and sepsis in June 1995. This case was considered to be consistent with atypical CML (aCML) according to the FAB classification because monocytosis was not observed. It seems likely and interesting that the coexistent monoclonal gammopathy and aCML might have arisen from common abnormal hematopoietic stem cells.
Aged↗
Corneal wound healing in tenascin knockout mouse.
PURPOSE: Tenascin (TN) is a large hexameric extracellular matrix glycoprotein that is expressed in developing organs and tumors. It has also been reported that TN is expressed in the embryonic cornea and during corneal wound healing. However, the role of TN in the cornea is not fully known. In this study, the role of TN in corneal wound healing was examined using the TN knockout (KO) mouse. METHODS: Two different injuries (a linear perforation wound and two 10-0 nylon suture wounds) were made separately on the corneas of both TNKO and congenic wild-type mice. The corneal wound healing was compared histologically, and the expression of TN and fibronectin (FN) on the injured cornea was examined immunohistochemically and by immunoblot analysis. RESULTS: Based on histologic analysis, there was no significant difference in the wound healing process between wild-type and TNKO mice in the linear incision experiment. However, the corneal stromata of TNKO mice were compressed prominently and devoid of migrating keratocytes in suture injury, which induced a more significant amount of TN than perforation wounds. Although FN expression on the sutured corneas of TNKO mice was upregulated during suture injury, the amount of FN protein was smaller than that of wild-type mice at the same time points after injury. CONCLUSIONS: In suture wounds, TN appears to enhance the amount of FN expression, and a lack of TN may impair stromal cell migration. TN plays a significant role in corneal wound healing, especially for wounds with mechanical stress.
Animals↗