Detection of acceptor sites for antisense oligonucleotides on native folded RNA by fluorescence-labeled oligonucleotide.

Pyrene-labeled 2'-O-methyloligoribonucleotide (OMUpy) and 5'-Ru(II) complex labeled oligodeoxyribonucleotie (Ru-probe) were prepared as the fluorescence probes to detect acceptor sites of antisense molecules on native folded RNA under the physiological condition. OMUpy showed the remarkable increase of the fluorescence intensity (334-fold at 375 nm) only when hybridized with complementary oligoRNA. When OMUpy was applied to E.

Synthesis of antisense oligonucleotides containing photocleavable protecting groups on the thymine bases and their photoinduced duplex formation.

Oligonucleotides containing photocleavable protecting groups at thymine bases were synthesized to induce the duplex formation by photo-irradiation. 6-Nitroveratryloxycarbonyl (NVOC) group was used for the photocleavable protecting group at N3 position of thymidine. An oligonucleotide containing NVOC groups (NVOC-ODN2:5'-dATG CAC CAT(NVOC) TCT(NVOC)GTC TGT-3') was synthesized by phosphoramidite method.

Suppression of dextran sodium sulfate-induced colitis in mice by zerumbone, a subtropical ginger sesquiterpene, and nimesulide: separately and in combination.

Ulcerative colitis (UC) and Crohn's disease are inflammatory disorders of unknown cause and difficult to treat, though some synthetic chemicals, including ligands for peroxisome proliferator-activated receptors (PPARs), are anticipated to be useful drugs. In contrast, few food phytochemicals have been reported to suppress colitis in animal models. The present study was undertaken to explore the suppressive efficacy of zerumbone (ZER), a sesquiterpenoid present in the rhizome of Zingiber zerumbet Smith that is used as a condiment in Southeast Asian countries and known to be a potent suppressant of cyclooxygenase (COX)-2 and inducible nitric oxide synthase expression in cell culture systems.

Selection of RNA-binding peptides containing Arg-rich motif.

In order to search the RNA-binding peptides, selection from randomized peptides containing Arg-rich motif was carried out using rRNA-immobilized column. As a result of the selection for 16S- and 23S-rRNA from E. coli, six peptides were identified to be the rRNA-binding peptide by MALDI-TOF MS.

Effect of chemical features and stability of polyplexes on their gene transfer efficiency.

Chemically modified poly-L-lysine (PL) derivatives with two essential features, which we have recently reported on, have been used to study key factors affecting transgene expression efficiency. PL derivatives having both of N epsilon-trimethyl lysine residue and 25 mole % serine residue showed enhanced transfection efficiency. When PL was modified in either way, no marked enhancement in gene expression was observed.

Identification of novel mutations of the CHST6 gene in Vietnamese families affected with macular corneal dystrophy in two generations.

Purpose: To report the clinical and genetic findings of Vietnamese families affected with macular corneal dystrophy (MCD) in 2 generations.

Methods: Two families, including 7 patients and 3 unaffected members, were examined clinically. Blood samples were collected.

Mutation analysis of the carbohydrate sulfotransferase gene in Vietnamese with macular corneal dystrophy.

Purpose: Mutations in a new carbohydrate sulfotransferase gene (CHST6) encoding corneal N-acetylglucosamine-6-sulfotransferase (C-GlcNac-6-ST) have been identified as the cause of macular corneal dystrophy (MCD) in various ethnicities. This study was conducted to examine the CHST6 gene in Vietnamese with MCD.

Methods: Nineteen unrelated families, including 35 patients and 38 unaffected relatives were examined clinically.

Mutations in the CHST6 gene in patients with macular corneal dystrophy: immunohistochemical evidence of heterogeneity.

Purpose: Macular corneal dystrophy (MCD) is an autosomal recessive disorder leading to severe visual impairment. The carbohydrate sulfotransferase 6 (CHST6) gene, which encodes the corneal N-acetylglucosamine 6-O-sulfotransferase on 16q22 has been identified as a causative gene for MCD. The purpose of this study was to identify mutations in CHST6 in Japanese patients with MCD and evaluate them by means of immunohistochemistry.

OPA1 gene mutations in Japanese patients with bilateral optic atrophy unassociated with mitochondrial DNA mutations at nt 11778, 3460, and 14484.

Purpose: To report mutations in the OPA1 gene in Japanese patients with bilateral optic atrophy unassociated with mitochondrial DNA mutations at nt 11778, 3460, and 14484.

Methods: Twelve unrelated patients with bilateral optic atrophy and 100 healthy controls were examined. Each exon of the OPA1 gene was amplified by polymerase chain reaction (PCR).

Psoralen-conjugated oligonucleotide with hairpin structure as a novel photo-sensitive antisense molecule.

Hairpin type psoralen-cojugated oligonucleotides cross-linked with RNA when they hybridized with a perfectly complementary RNA.

Combinatorial effects of nonsteroidal anti-inflammatory drugs and food constituents on production of prostaglandin E2 and tumor necrosis factor-alpha in RAW264.7 murine macrophages.

Combinatorial chemopreventive strategies, in contrast to those with individual agents, show potential in terms of potentially lower toxicity and higher efficacy. In this study, we combined several agents and examined their suppressive effects on the combined lipopolysaccharide (LPS)- and interferon(IFN)-gamma-induced formation of proinflammatory mediators, including prostaglandin (PG) E2 and tumor necrosis factor (TNF)-alpha, in RAW264.7 murine macrophages.

Detection of acceptor sites for antisense oligonucleotides on native folded RNA by fluorescence spectroscopy.

Antisense strategy has high potential for curing diseases and studying gene functions by suppressing the translation step. For the strategy, it is essential to detect acceptor sites of antisense molecules on mRNA under physiological conditions. We propose a new analytical method for the detection of acceptor sites of antisense molecules with high sensitivity.

Multiplex polymerase chain reaction for detection of herpes simplex virus type 1, type 2, cytomegalovirus, and varicella-zoster virus in ocular viral infections.

Purpose: To detect simultaneously herpes simplex virus type 1 (HSV-1), type 2 (HSV-2), cytomegalovirus (CMV), and varicella-zoster virus (VZV) in ocular specimens suspected of indicating viral infection, and to compare the results of multiplex polymerase chain reaction (PCR) with those of uniplex PCR.

Methods: Forty specimens, collected from 33 patients with clinically suspected herpes virus ocular infection, were tested. DNA was extracted from the specimens and amplified by multiplex and uniplex PCR.

A novel mutation of the TGFBI gene found in a Vietnamese family with atypical granular corneal dystrophy.

Background: Mutation of the human transforming growth factor beta-induced (TGFBI) gene causes granular corneal dystrophy (GCD) in various ethnic groups. In this report, we identify the genetic defect on the TGFBI gene in a Vietnamese family with atypical GCD .

Cases: The patient and her relatives were examined clinically.

Effects of selected food factors with chemopreventive properties on combined lipopolysaccharide- and interferon-gamma-induced IkappaB degradation in RAW264.7 macrophages.

Degradation of IkappaB (IkappaB) is a key step for nuclear factor-kappaB (NF-kappaB)-induced transcription of certain proinflammatory genes, including inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. We selected seven chemopreventive agents and examined their effects on combined lipopolysaccharide- and interferon-gamma-induced IkappaB degradation in RAW264.7 murine macrophages.

Search for Naturally-occurring Antioxidative Chemopreventors on the Basis of the Involvement of Leukocyte-derived Reactive Oxygen Species in Carcinogenesis.

Chemoprevention with food phytochemicals is currently regarded as one of the most attractive strategies for cancer control. We have been continuously working on the identification and characterization of chemopreventive phytochemicals extracted from a diverse range of edible plants. Recently, we have utilized a convenient assay, the tumor promoter-induced superoxide generation test, with differentiated HL-60 cells for primary screening, and performed chemical studies of antioxidative food phytochemicals.

Suppressive Effects of Edible Thai Plants on Superoxide and Nitric Oxide Generation.

We screened ethanol extracts from a total of 134 species of edible Thai plants for their suppressive effects on superoxide (O2(-)) generation using a xanthine (XA)-xanthine oxidase (XOD) assay system. When the extracts were tested at a concentration of 500 &mgr;/g/ml, 28.4% significantly suppressed O2(-) generation.

Synergistic suppression of superoxide and nitric oxide generation from inflammatory cells by combined food factors.

In contrast to chemopreventive strategies using individual agents, a combination of specified compounds may be effectual to achieve desirable results with higher efficacy and lower toxicity. In the present in vitro study, we examined combinations of agents and assessed which concentrations were appropriate to yield notable synergism. L-N(G)-Monomethyl-L-arginine (L-NMMA), a synthetic inducible nitric oxide synthase (iNOS) inhibitor, and zerumbone, a natural sesquiterpene that suppresses iNOS de novo synthesis, were combined at various concentrations, with the aim to diminish combined lipopolysaccharide- and interferon-gamma-induced nitric oxide generation in a murine macrophage line, RAW264.

A phase II detoxification enzyme inducer from lemongrass: identification of citral and involvement of electrophilic reaction in the enzyme induction.

We have developed a simple system for the sensitive detection and measurement of glutathione S-transferase (GST) activity that detoxifies polycyclic aromatic hydrocarbons using the cultured rat normal liver epithelial cell line, RL34 cells. Citral (3,7-dimethyl-2,6-octadienal) was isolated from the methanol extract of lemongrass (Cymbopogon citratus) and identified as a novel inducer of GST. Citral, a mixture of the two stereoisomers geranial and neral, dose- and time-dependently induced the total and pi-class-specific activities of GST.

A novel mutation of M1S1 gene found in a Vietnamese patient with gelatinous droplike corneal dystrophy.

To identify the genetic defect in the M1S1 gene responsible for gelatinous droplike corneal dystrophy (GDLD) in a Vietnamese family.Experimental study. Blood samples were collected from a patient and the unaffected members of a GDLD-affected family.

Mutagenicity of reactive oxygen and nitrogen species as detected by co-culture of activated inflammatory leukocytes and AS52 cells.

Activated inflammatory leukocytes generate a variety of reactive oxygen and nitrogen species (RONS) that may have roles in mutagenesis and carcinogenesis. The purpose of the present study was to explore the relationship between inflammatory leukocyte activation and mutagenesis using co-culture systems. We investigated the mutagenic potentials of 12-O-tetradecanoylphorbol-13-acetate (TPA)-stimulated differentiated HL-60 (human promyelocytic leukemia cells), and RAW 264.

Heterozygous Ala137Pro mutation in keratin 12 gene found in Japanese with Meesmann's corneal dystrophy.

Purpose: To report the molecular genetic analysis of a Japanese pedigree with Meesmann's corneal dystrophy (MCD).

Methods: Sequencing of the keratin 3 and keratin 12 genes was performed in 2 patients who were siblings and in an unaffected individual in the same family. The patients had the typical corneal microcysts and recurrent erosions with mild photophobia.

Identification of a stable chymase inhibitor using a pharmacophore-Based database search.

In general, serine protease chymase inhibitors readily decompose in plasma. We previously found that thiazolidine-2,4-dione and thiadiazole derivatives are also unstable. Using a pharmacophore-based database search, we identified a benzo[b]thiophen-2-sulfonamide derivative as a stable chymase inhibitor.

Further investigation of the modifying effect of various chemopreventive agents on apoptosis and cell proliferation in human colon cancer cells.

Purpose: Recent preclinical assays using animal models have shown that naturally-occurring and synthetic chemicals such as auraptene (AUR), nobiletin (NOB), hesperidin (HE), diosmin (DIO), indole-3-carbinol (I3C), 1'-acetoxychavicol acetate (ACA), 2,5-di-O-acetyl-D-1,4-glucaro-6,3-dilactone (ACE), D-glucuronic acid gamma-lactone (GL), chlorogenic acid (CGA), protocatechuic acid (PA), and sinigrin (SIN) are possible preventive agents against the development of cancer. However, the mode of action of such preventive agents remains to be elucidated. The current study, therefore, was conducted to analyze whether these agents induce apoptosis and/or inhibit DNA synthesis in human colorectal cancer cell lines.