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1.
J Biomed Sci ; 22: 64, 2015 Aug 04.
Article in English | MEDLINE | ID: mdl-26239837

ABSTRACT

BACKGROUND: Osteoarthritis (OA) is a common joint disease that causes disabilities in elderly. However, few agents with high efficacy and low side effects have been developed to treat OA. In this study, we evaluated the effects of the alginate extract named CTX in OA cell and rabbit models. RESULTS: CTX was formulated by hydrolyzing sodium alginate polymers with alginate lyase and then mixing with pectin. HPLC was used to analyze the CTX content. Human chondrosarcoma SW1353 cells treated with interleukin-1ß were used as OA model cells to investigate the effects of CTX on chondrocyte inflammation and anabolism. CTX at concentrations up to 1000 µg/ml exerted low cytotoxicity. It inhibited the gene expression of proinflammatory matrix metalloproteinases (MMPs) including MMP1, MMP3 and MMP13 in a dose-dependent manner and increased the mRNA level of aggrecan, the major proteoglycan in articular cartilage, at 1000 µg/ml. Thirteen-week-old New Zealand White rabbits underwent a surgical anterior cruciate ligament transection and were orally treated with normal saline, glucosamine or CTX for up to 7 weeks. Examinations of the rabbit femur and tibia samples demonstrated that the rabbits taking oral CTX at a dosage of 30 mg/kg/day suffered lesser degrees of articular stiffness and histological cartilage damage than the control rabbits. CONCLUSIONS: The gene expression profiles in the cell and the examinations done on the rabbit cartilage suggest that the alginate extract CTX is a pharmaco-therapeutic agent applicable for OA therapy.


Subject(s)
Alginates/administration & dosage , Chondrocytes/drug effects , Osteoarthritis/drug therapy , Pectins/administration & dosage , Polysaccharide-Lyases/administration & dosage , Administration, Oral , Alginates/chemistry , Animals , Cartilage, Articular/drug effects , Cartilage, Articular/pathology , Chondrocytes/pathology , Gene Expression Regulation/drug effects , Glucuronic Acid/administration & dosage , Glucuronic Acid/chemistry , Hexuronic Acids/administration & dosage , Hexuronic Acids/chemistry , Humans , Interleukin-1beta/toxicity , Matrix Metalloproteinase 1/biosynthesis , Matrix Metalloproteinase 13/biosynthesis , Matrix Metalloproteinase 3/biosynthesis , Osteoarthritis/chemically induced , Osteoarthritis/pathology , Pectins/chemistry , Polysaccharide-Lyases/chemistry , Rabbits
2.
J Microbiol Biotechnol ; 25(5): 718-22, 2015 May.
Article in English | MEDLINE | ID: mdl-25502828

ABSTRACT

Techniques for immobilizing effective enzymes on nanoparticles for stabilization of the activity of free enzymes have been developing as a pharmaceutical field. In this study, we examined the effect of three different pH conditions of phosphate buffer, as a dissolving solvent for lysosomal enzymes, on the direct immobilization of lysosomal enzymes extracted from Hen's egg white and Saccharomyces cerevisiae. Titanium(IV) oxide (TiO2) nanoparticles, which are extensively used in many research fields, were used in this study. The lysosomal enzymes immobilized on TiO2 under each pH condition were evaluated to maintain the specific activity of lysosomal enzymes, so that we can determine the degree of melanin treatment in lysosomal enzymes immobilized on TiO2. We found that the immobilization efficiency and melanin treatment activity in both lysosomal enzymes extracted from Hen's egg white and S. cerevisiae were the highest in an acidic condition of phosphate buffer (pH 4). However, the immobilization efficiency and melanin treatment activity were inversely proportional to the increase in pH under alkaline conditions. In addition, enhanced immobilization efficiency was shown in TiO2 pretreated with a divalent, positively charged ion, Ca(2+), and the melanin treatment activity of immobilized lysosomal enzymes on TiO2 pretreated with Ca(2+) was also increased. Therefore, this result suggests that the immobilization efficiency and melanin treatment activity of lysosomal enzymes can be enhanced according to the pH conditions of the dissolving solvent.


Subject(s)
Enzymes, Immobilized/chemistry , Lysosomes/chemistry , Melanins/metabolism , Titanium/chemistry , Animals , Calcium/metabolism , Chickens , Enzymes, Immobilized/metabolism , Hydrogen-Ion Concentration , Lysosomes/metabolism , Melanins/chemistry , Solvents
3.
J Agric Food Chem ; 60(16): 4053-9, 2012 Apr 25.
Article in English | MEDLINE | ID: mdl-22352834

ABSTRACT

For the identification of a novel insecticidal protein, a two-dimensional liquid chromatography (PF-2D) system was used in a quantitative proteomic analysis of Xenorhabdus nematophila CBNU strain isolated from entomophagous nematode Steinernema carpocapsae . Protein patterns obtained from minimum and maximum insecticidal activities during cultivation were contrasted, and a novel toxin protein (Txp40) was identified by MALDI-TOF/MS. The DNA sequence of the cloned toxin gene (1089 bp) has an open reading frame encoding 363 amino acids with a predicted molecular mass of 41162 Da. The txp40 identified in this study is most closely related to the known txp40 cloned from X. nematophila EB (ADQ92844) with 94.4% identical sequence residues. Following the expression of the newly identified toxin gene in Escherichia coli , the insecticidal activity of the recombinant toxin protein was determined against Plutella xylostella larvae; a 56.7% mortality rate was observed within 24 h.


Subject(s)
Bacterial Proteins/genetics , Bacterial Toxins/genetics , Insecticides/metabolism , Moths/drug effects , Proteomics , Rhabditida/microbiology , Xenorhabdus/metabolism , Amino Acid Sequence , Animals , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Bacterial Proteins/pharmacology , Bacterial Toxins/chemistry , Bacterial Toxins/metabolism , Bacterial Toxins/pharmacology , Insecticides/chemistry , Insecticides/pharmacology , Larva/drug effects , Larva/growth & development , Molecular Sequence Data , Moths/growth & development , Sequence Alignment , Xenorhabdus/chemistry , Xenorhabdus/genetics , Xenorhabdus/isolation & purification
4.
Environ Sci Technol ; 43(24): 9335-40, 2009 Dec 15.
Article in English | MEDLINE | ID: mdl-20000526

ABSTRACT

Single-stranded DNA aptamers were generated from a random library to remove arsenic from Vietnamese groundwater. On the basis of significant arsenic contamination levels, three areas in Ha Nam province (Vinh Tru, Bo De, and Hoa Hau) and five areas near the Mekong River Delta (MR1-5) were selected as study areas. The aptamers were in vitro selected using an arsenic aptamer affinity column created by immobilizing arsenic on Affi-gel 10 resin. Quantitative analyses of the aptamer candidates Ars-1 to Ars-8 by surface plasmon resonance (SPR) revealed the Ars-3 aptamer to have the highest affinity to arsenate [(As(V)] and arsenite [As(III)] with a dissociation constant (K(d)) of 4.95 +/- 0.31 and 7.05 +/- 0.91 nM, respectively. The specific affinity interactions of the Ars-3 aptamer to arsenic were verified against other heavy metals. After obtaining successful removal results with a laboratory-prepared aqueous arsenic solution, Ars-3 was applied for removal of any arsenic present in the groundwater samples collected from the studied areas in Vietnam. Field results were also successful: various arsenic concentrations ranging from 28.1 to 739.2 microg/L were completely removed after 5 min of incubation with the arsenic-binding aptamer Ars-3.


Subject(s)
Aptamers, Nucleotide/chemistry , Arsenic/chemistry , Water Pollutants, Chemical/chemistry , Water Purification/methods , Water Supply/analysis , Humans , Molecular Structure , Nucleic Acid Conformation , Rivers/chemistry , SELEX Aptamer Technique , Surface Plasmon Resonance/methods , Vietnam , Water Purification/instrumentation
5.
Water Res ; 37(5): 1091-9, 2003 Mar.
Article in English | MEDLINE | ID: mdl-12553984

ABSTRACT

Electrodialysis experiments of the lysine fermentation waste were performed to generate demineralized feed and ammonium sulfate, which can be utilized as a fertilizer and an animal feed, respectively. The electrodialysis performances were compared for different ion exchange membranes in terms of ammonium sulfate removal rate, resistance and conductivity change. Analysis of fouling phenomena revealed that organics fouled ion exchange membranes reversibly in electrodialysis of the fermentation waste. In this study, mitigation of membrane fouling with the pulsed electric field was examined for the electrodialysis of the fermentation waste containing strong foulants. The half-wave power reduced membrane fouling significantly. For a quantitative measure of the membrane fouling tendency, a membrane fouling index for electrodialysis was used. This study showed the potential use of pulsed power as an effective fouling mitigation method for the electrodialysis of fermentation waste.


Subject(s)
Ammonium Sulfate/isolation & purification , Lysine/metabolism , Refuse Disposal/methods , Ammonium Sulfate/chemistry , Animal Feed , Conservation of Natural Resources , Electrochemistry , Fermentation , Fertilizers , Ion Exchange , Membranes, Artificial
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