Park, Hyung-Seok;Hwang, Ji-Young;Shin, Ueon-Sang;Kim, Hae-Won;Gong, Myoung-Seon
Bulletin of the Korean Chemical Society
/
v.32
no.10
/
pp.3581-3586
/
2011
Silver nanoparticle (Ag-NPs)-immobilized and amine-functionalized carbon nanotubes (MWCNTs), MWCNT-Ag-$NH_2$, were easily prepared in order to develop an efficient delivery system of biomolecules without complicated processes of manufacture. For this, Ag-NPs-immobilized MWCNTs, MWCNT-Ag, were initially prepared in order to create large surface area to enable more efficient linkage with guest-molecules using pristine MWCNTs. The Ag-NPs on MWCNTs were further positively functionalized with 2-aminoethanthiol to allow ionic linkage with biomolecules. Ultimately, the positively charged delivery system proved to be highly effective for the binding capacity of bovine serum albumin (BSA) as a negatively charged model protein, when compared to that of lysozyme used as a positively charged model protein. The releasing profile of BSA was observed in almost linear pattern for about two weeks in a saline solution. This study demonstrated the potential usefulness of the pristine MWCNTs in conjunction with Ag-NPs for the selective delivery of many (negatively or positively) charged biomolecules including proteins and genes.
This study was conducted to investigate the noodle added with Pachyrhizus erosus powder. Total contents of carbohydrates, crude protein, crude lipid, ash, and total dietary fiber were 87.14%, 3.14%, 0.81%, 7.82%, and 49.12%, respectively. The water-binding capacity was significantly increased by increasing amount of the Pachyrhizus erosus powder. When the volume of the noodles was measured after cooking, there was no difference between the control and Pachyrhizus erosus powder groups. The texture characteristics of hardness increased by increasing amount of the Pachyrhizus erosus powder. As a result of the sensory test, the 1.5% Pachyrhizus erosus powder noodles showed the highest scores for all items including appearance, flavor, taste, texture and overall acceptability. The bacterial counts of noodle decreased with greater Pachyrhizus erosus powder content on the third days. It is considered that research on antimicrobial activity should proceed in the future. As a result, it was finally suggested that optimum level of the Pachyrhizus erosus powder in the product was 1.5% level.
Kim, Kwan;Kang, Kil-Jin;Lee, Yong-Hyun;Kim, Sung-Kon
Korean Journal of Food Science and Technology
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v.25
no.1
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pp.86-87
/
1993
The steeping of waxy rice in water at $20^{\circ}C$ for 12 hr resulted in the decrease of protein, crude fat and ash contents. Water-binding capacity and swelling power (at $80^{\circ}C$) of waxy rice flour increased upon steeping. Maximum viscosity of waxy rice flour in 0.19 N sodium hydroxide solution was increased up to 6 hr steeping and decreased thereafter. The amylograph indices increased as the steeping time increased. The degree of changes in properties during steeping was different between two waxy rice cultivars.
A library of unlimited number of novel lectins with diverse specificities has been previously generated by randomly mutating the carbohydrate-recognition domain of Maackia amurensis hemagglutinin (MAH). To establish the experimental environment capable of selecting high affinity mutant lectins in E. coli, phage display system was adapted. Carbohydrate binding capacity of two phagemid vectors, pComb3 and pComb8 displaying wild-type MAH lectin was assessed. Specific bindings of pComb3 and pComb8 phages expressing w.t. MAH to affinity-purified polyclonal anti-MAH antibody and to glycophorin was demonstrated. Both phages also showed strong hemagglutinating activity to intact but not sialidase-treated human erythrocytes, which is consistent to the specificity of native MAH. Taken together, two different phage display vectors successfully allowed the expression of active MAH as a fusion protein on the surface of bacteriophage, which will lead to preparation of unique plant lectins with high affinity toward a variety of carbohydrate chains.
Objective: Unlike mammals, goose fatty liver shows a strong tolerance to fatty acids without obvious injury. Stearyl-coenzyme A desaturase 1 (SCD1) serves crucial role in desaturation of saturated fatty acids (SAFs), but its role in the SAFs tolerance of goose hepatocytes has not been reported. This study was conducted to explore the role of SCD1 in regulating palmitic acid (PA) tolerance of goose primary hepatocytes. Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide was examined to reflect the effect of PA on hepatocytes viability, and quantitative polymerase chain reaction was used to detect the mRNA levels of several genes related to endoplasmic reticulum (ER) stress, inflammation, and apoptosis, and the role of SCD1 in PA tolerance of goose hepatocytes was explored using RNA interfere. Results: Our results indicated that goose hepatocytes exhibited a higher tolerant capacity to PA than human hepatic cell line (LO2 cells). In goose primary hepatocytes, the mRNA levels of fatty acid desaturation-related genes (SCD1 and fatty acid desaturase 2) and fatty acid elongate enzyme-related gene (elongase of very long chain fatty acids 6) were significantly upregulated with 0.6 mM PA treatment. However, in LO2 cells, expression of ER stress-related genes (x box-binding protein, binding immunoglobulin protein, and activating transcription factor 6), inflammatory response-related genes (interleukin-6 [IL-6], interleukin-1β [IL-1β], and interferon-γ) and apoptosis-related genes (bcl-2-associated X protein, b-cell lymphoma 2, Caspase-3, and Caspase-9) was significantly enhanced with 0.6 mM PA treatment. Additionally, small interfering RNA (siRNA) mediated downregulation of SCD1 significantly reduced the PA tolerance of goose primary hepatocytes under the treatment of 0.6 mM PA; meanwhile, the mRNA levels of inflammatory-related genes (IL-6 and IL-1β) and several key genes involved in the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), forkhead box O1 (FoxO1), mammalian target of rapamycin and AMPK pathways (AKT1, AKT2, FoxO1, and sirtuin 1), as well as the protein expression of cytochrome C and the apoptosis rate were upregulated. Conclusion: In conclusion, our data suggested that SCD1 was involved in enhancing the PA tolerance of goose primary hepatocytes by regulating inflammation- and apoptosis-related genes expression.
The optic nerve often suffers regenerative failure after injury, leading to serious visual impairment such as glaucoma. The main inhibitory factors, including Nogo-A, oligodendrocyte myelin glycoprotein, and myelin-associated glycoprotein, exert their inhibitory effects on axonal growth through the same receptor, the Nogo-66 receptor (NgR). Oncomodulin (OM), a calcium-binding protein with a molecular weight of an ~12 kDa, which is secreted from activated macrophages, has been demonstrated to have high and specific affinity for retinal ganglion cells (RGC) and promote greater axonal regeneration than other known polypeptide growth factors. Protamine has been reported to effectively deliver small interference RNA (siRNA) into cells. Accordingly, a fusion protein of OM and truncated protamine (tp) may be used as a vehicle for the delivery of NgR siRNA into RGC for gene therapy. To test this hypothesis, we constructed OM and tp fusion protein (OM/tp) expression vectors. Using the indirect immunofluorescence labeling method, OM/tp fusion proteins were found to have a high affinity for RGC. The gel shift assay showed that the OM/tp fusion proteins retained the capacity to bind to DNA. Using OM/tp fusion proteins as a delivery tool, the siRNA of NgR was effectively transfected into cells and significantly down-regulated NgR expression levels. More importantly, OM/tp-NgR siRNA dramatically promoted axonal growth of RGC compared with the application of OM/tp recombinant protein or NgR siRNA alone in vitro. In addition, OM/tp-NgR siRNA highly elevated intracellular cyclic adenosine monophosphate (cAMP) levels and inhibited activation of the Ras homolog gene family, member A (RhoA). Taken together, our data demonstrated that the recombinant OM/tp fusion proteins retained the functions of both OM and tp, and that OM/tp-NgR siRNA might potentially be used for the treatment of optic nerve injury.
The study was performed to determine the effect of faba bean protein isolate (FBPI) alone or in combination with microbial transglutaminase (MTG) on the rheological properties of pork myofibrillar protein gel (MPG), and physiochemical and textural properties of reduced-salt, low-fat pork model sausages (LFMSs). The cooking yields of MPGs with MTG or FBPI alone decreased and increased, respectively. However, the combination of FBPI and MTG was similar to the control (CTL) without FBPI or MTG. Gel strength values of MPG added with both FBPI and MTG were higher than treatments with FBPI or MTG alone. The hydrophobicity values of CTL were lower than those of MPG with FBPI alone, whereas the addition of MTG decreased the hydrophobicity of MPGs. The incorporation of FBPI alone or in combination with MTG decreased sulfhydryl groups (p<0.05). Shear stress values of MPGs with MTG tended to be higher than those of non-MTG treatments at all shear rates, and the addition of FBPI into MPGs increased shear stress values. Reduced-salt (1.0%) LFMSs with FBPI alone or combined with MTG had both lower cooking loss and expressible moisture values than those of CTL and similar values to the reference sample (REF, 1.5% salt). Textural properties of reduced-salt LFMSs with FBPI or MTG were similar to those of REF. These results demonstrated that the combination of FBPI and MTG could improve the water binding capacity and textural properties of pork MPGs and LFMSs and might be suitable for application in the development of healthier meat products.
Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.
We investigated the binding properties of $(^3H)$ QNB and $(^3H)$ NMS to mAchR to elucidate the characterstics of mAchR in rat brain by using two different preparations (homogemates & intact brain cell aggregates). The binding properties of both ligands demonstrated high affinity and saturability in both experiments, however $(^3H)$ QNB showed a significantly higher maximal binding capacity than tha ot $(^3H)$ NMS 1. In rat brain homogenates; Displacement of both lignands with several mAchR antagonists resulted in competition curves in accoradnce with the law of massaction for QNB, atropine & scopolamine in thie preparation, also a similar profile was found for the quaternary ammonium analogs of atropine & scopolamine (methyl atropine & methylscopolamine) when $(^3H)$ NMS was used to label the receptors in rat brain. But when these hydrophillic antagonists were used to displace $(^3H)$ QNB, they showed interaction with high- and low-affinity binding sites in brain homogenates. Pirenzepine, the nonclassical mAchR antagonist, was able to displace both ligands from binding sites in this preparation. 2. In intact rat brain cell aggregates; Intact bain cell aggregates were used to elucidate the binding characteristics of $(^3H)$ NMS to mAchR in rat. The magnitude of binding of this ligand was related linearly to the amount of cell protein in the binding assay with a high ratio of total to nonspecific binding. mAchR antagonists displaced specific $(^3H)$ NMS binding according to the law of mass-action, while it was possible to resolve displacement curves using mAchR agonist into high-& low-affinity component. 3. Our results indicate that more hydrophilic receptor ligand $(^3H)$ QNB, displacement experiments in both tissues demonstrated that the lipid solubility of a particulr mAchR ligand might play an important role in determining its profile of binding to the mAchR, and the concentrations of mAchR in rat brain are both on the cell surface (membrane-bound receptor) and in the intracelluar membrane (intermembrane-bound receptor). 4. The results are discussed in terms of the usefulness of dissociated intact rat brain cells in studying mAchR in central nervous system.
Physicochemical properties and storage stability of plant-based alternative meat prepared with low-fat soybean powder (LPAM) treated by supercritical-CO2 and those of full-fat soybean powder (FPAM) were compared. Ash and crude protein contents were higher in LPAM than in FRAM. Water absorption capacity and oil absorption capacity were significantly higher in LPAM than in FPAM. Water binding capacity was higher in LPAM than in FPAM during a 20 days storage period at 5℃ and pH was significantly lower in LPAM than in FPAM after a 5~10 days storage period. Hardness, gumminess and chewiness significantly increased with the increase in the storage period, and the three were significantly higher in LPAM than in FPAM after 10 days and 20 days of storage. The acid value showed no remarkable difference according to the storage period in LPAM; however, it was significantly higher in FPAM than in LPAM after 20 days of storage. The peroxide value and TBA value were significantly increased according to the storage period, and were significantly lower iin LPAM than in FPAM during all the storage periods. Therefore, the use of low-fat soybean powder may be effective in improving oxidative stability during storage in the production of plant-based alternative meat.
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