Apoptotic Effects of Anthocyanins from Vitis coignetiae Pulliat Are Enhanced by Augmented Enhancer of the Rudimentary Homolog (ERH) in Human Gastric Carcinoma MKN28 Cells

Evidence suggests that augmented expression of a certain gene can influence the efficacy of targeted and conventional chemotherapies. Here, we tested whether the high expression of enhancer of the rudimentary homolog (ERH), which serves as a prognostic factor in some cancers, can influence the efficacy of anthocyanins isolated from fruits of Vitis coignetiae Pulliat, Meoru in Korea (AIMs) on human gastric cancer cells. The anticancer efficacy of AIMs was augmented in ERH-transfected MKN28 cells (E-MKN28 cells). Molecularly, ERH augmented AIM-induced caspase-dependent apoptosis by activating caspase-3 and -9. The ERH-augmented apoptotic effect was related to mitochondrial depolarization and inhibition of antiapoptotic proteins, XIAP, and Bcl-2. In addition, reactive oxygen species (ROS) generation was augmented in AIMs-treated E-MKN28 cells compared to AIMs-treated naïve MKN28 cells. In conclusion, ERH augmented AIM-induced caspase-dependent mitochondrial-related apoptosis in MKN28 cells. A decrease in expression of Bcl-2 and subsequent excessive ROS generation would be the mechanism for ERH-augmented mitochondrial-related apoptosis in AIMs-treated MKN28 cells. A decrease in expression of XIAP would be another mechanism for ERH-augmented caspase-dependent apoptosis in AIMs-treated MKN28 cells.     

Functional characterization of purified pear protease and its proteolytic activities with casein and myofibrillar proteins

This study was performed to characterize pear protease proteolytic activity and investigate the use of pear protease as a meat tenderizer. Pear protease was purified and stabilized by 5% dextrin during lyophilization (dry) or concentration (liquid). Pear protease was further characterized with respect to pH, thermodynamics, and enzyme kinetics. Pear protease was stable at a pH range of 5-8 with an optimum pH of 6.5. From Arrhenius plots, liquid protease showed higher temperature dependency (23.49 kJ/mol) than dry protease (18.62 kJ/mol) due to its higher activation energy. The kcat/Km, catalytic efficiency of enzyme, was similar with 2.9 and 2.7 µM/min with dry and liquid proteases. Pear protease was evaluated for its proteolytic activities with casein and beef myofibrillar proteins by individually and combination with fig and kiwifruit proteases. These result indicated that pear and kiwifruit proteases could be complementary to be a desirable product for meat tenderization.     

Biochemical monitoring of black raspberry (Rubus coreanus Miquel) fruits according to maturation stage by H NMR using multiple solvent systems

Nuclear magnetic resonance (NMR) techniques coupled with multivariate data analysis were used to conduct monitoring of biochemical changes of black raspberry fruits at different stages of maturation and under various extraction and NMR dissolution solvent conditions: extraction with 50% methanol and D₂O as an NMR dissolution solvent, extraction with 50% methanol and 50% methanol-d₄ as an NMR dissolution solvent, and extraction with 100% ethyl acetate and 100% methanol-d₄ as an NMR dissolution solvent. Partial least-squares discriminant analysis reliably distinguished black raspberry fruits according to the maturation stage, whereby the relative levels of various compounds such as amino acids, organic acids, sugars and phenolic compounds were compared using analysis of variance. Sucrose and most of the amino acids, and organic acids decreased, whereas fructose, glucose, and cyanidins increased in relative concentration according to maturation of black raspberry fruits. The total number and kinds of assigned compounds of the three solvent systems were also compared. This research demonstrates that the metabolic profile of black raspberry fruits changes during maturation, and provides objective criteria for determining the stage of black raspberry fruit maturation via a ¹H NMR-based metabolomics technique using multiple solvent systems.     

Fabrication and magnetic characteristics of vertical feco nanowire arrayed in Al2O3 insulator of honeycomb bulkhead structure

High-density and uniform-sized FeCo alloy nanowires were prepared by electro deposition of Fe2+ and Co2+ into Anodic aluminum oxide (AAO) templates with two different diameters. These templates were fabricated with three-step anodization method. The additional anodization after the 2nd anodization step resulted in the decrease of the thickness of bottom barrier layer. It found an optimum condition to obtain a successful electrodeposition of Fe2+ and Co2+ into AAO templates. The nanowires with the diameters of 25 nm and 80 nm were homogeneously embedded in the AAO templates and their magnetic properties were strongly affected by diameters of nanowire.     

Relative distribution of free amino acids in buckwheat

The most abundant amino acid in the sprouts of common buckwheat (CB) was Val (40%), followed by Tyr (28%), whereas Val accounted for 62% in tatary buckwheat (TB). The buckwheat stem and root commonly contained Gln (40–42% in stem; 30–37% in root). Thus, soluble amino nitrogen source is used for Gln in buckwheat. The main difference of amino acid distribution in 3 tissues between CB and TB was Tyr in sprouts. A low level of Tyr in TB was presumably resulted from the conversion to other phenolic metabolites. The content of essential free amino acids in TB sprout was 53% higher than that in CB. Thus, the TB sprouts are beneficial to the human nutrition.     

Tensile strength and concrete cone failure in CFT connection with internal diaphragms

International Journal of Steel Structures - Concrete filled tube (CFT) system provides excellent structural performance thanks to the composite effect between the tube and the concrete. However,...     

Changes of the Cytoplasmic Proteome in Response to Alcoholic Hepatotoxicity in Rats

Proteomic analyses have already been used in a number of hepatological studies and provide important information. However, few reports have focused on changes in the cytoplasmic proteome. The present study therefore aimed to evaluate changes in cytoplasmic proteome of rats in response to alcoholic hepatotoxicity. Rats were fed a Liber-DeCarli liquid diet containing ethanol for four weeks. Cytoplasmic proteins except mitochondrial proteins from the livers of these animals were investigated using two-dimensional gel electrophoresis and mass spectrometry. Alcohol induced a decrease in body weight gain and an increase in alanine transaminase (ALT), cholesterol, and phospholipid levels. Histopathological observations revealed hepatic damage characterized by necrosis and fatty change in alcohol-treated group at week 2, which continues until week 4. Our proteomic analysis revealed that 25 proteins were differentially expressed in the ethanol-fed group. Of these, 12 cytoplasmic proteins are being reported for the first time. Taken together, our results provide further insights into the disease mechanism and therapeutic information of alcoholic liver disease.     

Assessing the digestibility of genetically modified soybean: Physiologically based in vitro digestion and fermentation model

Gastrointestinal digestibility of new proteins inserted in food supply is one of significant parameters for assessing safety of genetically modified (GM) foods on the assumption that digestive stability is undesirable. Digestive stability of GM soybean was evaluated under the physiologically based in vitro digestion model. Electrophoresis was used to follow the in vitro sequential digestions/fermentation procedure and Western blotting was employed to confirm the new protein in GM soybean.CP4EPSPS newly expressed in GM soybean was completely digested in 15 s in the gastric fluid in this model. In the assay of fermentation of new protein, CP4EPSPS was also complete degraded and no fermentation products were found during 24 h of colon fermentation. Results from this study suggested that CP4EPSPS in GM soybean does not have a potential allergenicity due to its complete digestion within a very short time in gastric fluid. No formation of colonic protein fermentation products confirms that new protein is not capable of initiating colon cancer.     

Affinity purification of 4-α-glucanotransferase through formation of complex with insoluble amylose

4-α-Glucanotransferases (αGTases) are useful enzymes to modify starch structure using their unique hydrolysis pattern and transglycosylation activity. In spite of lacking of a carbohydrate binding module, a thermostable aGTase from Thermus thermophilus (TTαGT) bound to insoluble amylose, leading to simple recovery of the enzyme-amylose complex using centrifugation. At a preparative scale, the recovery yield was 40.3% of the subjected free enzyme via collection of the complex twice. The complex exhibited improved thermostability, which shifted the optimum temperature from 70 to 80°C and increased the half-life at 90°C by three-fold compared with the free enzyme. Texture profile analysis of the gels made of modified starch by either free TTαGT or the complex revealed that the complex with double dosage showed the performance similar to free TTαGT in the modification of starch. In conclusion, the purification method described here would be useful due to easy scale-up and simple process without chromatographic process.     

Engineering interchain disulfide bonds into conserved framework regions of Fv fragments: improved biochemical characteristics of recombinant immunotoxins containing disulfide-stabilized Fv

Using molecular modeling technology, we have recently identifiedtwo positions in conserved framework regions of antibody Fvfragments (Fvs) that are distant from CDRs, and potentiallycan be used to make recombinant Fv fragments in which the unstableV_H and V_L heterodimer is stabilized by an interchain disulfidebond inserted between structurally conserved framework positions.A disulfide bond has been introduced at one of these positions,V_H44-V_L105, and shown to stabilize various Fvs that retain fullbinding and specificity. Recombinant immunotoxias, e.g. B3(dsFv)-PE38KDELin which this disulfide-stabilized Fv moiety is connected toa truncated form of Pseudomonas exotoxin (PE; PE38KDEL) whichcontains the translocation and ADP ribosylation domains, areindistinguishable in binding and specificity from its single-chainimmunotoxin counterparts. We have now analyzed the alternativeposition, (V_H111-V_L48), predicted by the modeling methodology,for disulfide stabilization of mAb B3(Fv) by producing a recombinantimmunotoxin with such disulfide-stabilized (ds) Fv. This immunotoxinwas also very active and retained full specificity to B3 antigen-positivecells. However, it was 2- to 3-fold less active than the V_H44-V_L105dsFv-molecule. We also tested various biochemical features ofV_H44-V_L105 and V_H111-V_L48 dsFv immunotoxins and compared themwith the corresponding single-chain immunotoxin. We found thedsFv immunotoxins were more stable in human serum and more resistantto thermal and chemical denaturation than the single chain (sc)Fv immunotoxin. Because dsFv immunotoxins and dsFvs have fullactivity and specificity and improved stability, they may bemore useful than scFv immunotoxins as therapeutic and diagnosticagents.