• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.362-370
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• 1995

Human plasma low density lipoprotein(LDL) is the main carrier for cholesterol, and recent studies suggest the normal LDL can be readily oxidized by free radical and not interact with LDL receptor. Lipoprotein pariticles are consisted of lipid andprotein, and fatty acids of lipoproteins are prone to oxidation. LDL particles readily undergo oxidative modification by copper. From the results, oxidized LDL altered its biological properties. A marked increase in the electrophoretic mobility of LDl on agarose gel indicated that negative surface charge of the LDL particles was increased. Also, the results from the HPLC showed that oxidized LDL was degraded into several polypeptides nonenzymatically. Degradation tests which measured the amount of 5-IAF labelled oxidized LDL were carried out by monocyte and hepatocyte cell culture. Hepatocyte cell culture of modified LDL did not show consistent pattern. However, binding rate of modified LDL with HMDM(human monocyte derived macrophage) was enhanced with oxidation, but was retarded by addition of antioxidants(hyaluronic acid, vitamin A, vitamin E). Also comparisons of oxidized-LDL, acetyl-LDL and MDA-LDL showed significant differences in the chemical properteis and binding affinity to HMDM. Thus, modificaition of normal LDL altered its biological properties.

• BMB Reports
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• v.29 no.6
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• pp.525-529
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• 1996

Ligand binding properties of muscarinic acetylcholine receptors (mAChRs) in the nematode Caenorhabditis elegans (C. elegans) were characterized by using filtration binding assays. Scatchard analysis using $[^{3}H]N-methylscopolamine$ ($[^{3}H]NMS$) showed that the dissociation constant ($K_d$) and the maximum binding value ($B_{max}$) were $3.3{\pm}0.8{\times}10^{10}$ M and $9.0{\pm}1.1$ fmol/mg protein, respectively. Binding competition experiments indicated that the affinities of C. elegans mAChRs to atropine, scopolamine, and oxotremorine were similar to those of mammalian mAChRs. Pirenzepine binding experiments revealed that the binding pattern of mAChRs in C. elegans closely resembled that of mAChRs in rat brain, suggesting that the receptors consist primarily of Ml subtype. The affinity of mAChRs for oxotrernorine was significantly affected by guanylylimidodiphosphate (Gpp(NH)p), a non hydrolyzable GTP analog, suggesting that mAChRs in C. elegans might be coupled to G proteins. The data presented here indicate the possibility that C. elegans provides a living animal model to study the action mode of the muscarinic cholinergic system.

• Journal of Life Science
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• v.26 no.5
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• pp.509-518
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• 2016

Asparagus cochinchinensis is a medical plant that has long been used to treat fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease in northeast Asian countries. Although several studies have been conducted on the anti-neuroinflammatory effects of A. cochinchinensis, the correlation between these effects and nerve growth factor (NGF) has not yet been examined. In this study, we investigated the effects of an aqueous extract of A. cochinchinensis (AEAC) on the secretion and action mechanism of NGF in neuronal cells. The concentration of the NGF protein in the supernatant collected from cultured cells increased significantly in B35 cells treated with AEAC in comparison with the vehicle-treated group without any specific cytotoxicity. Furthermore, the mRNA expression of NGF showed a very similar pattern to its protein concentration. To examine the bioactivity of NGF secreted from B35 cells, undifferentiated PC12 cells were cultured in an AEAC-conditioned medium and neuritic outgrowth was observed. The dendrite length of PC12 cells in the AEAC-treated group was significantly higher than that in the vehicle-treated group. Moreover, the level of the downstream effectors p-TrkA and p-ERK of the high-affinity NGF receptor was significantly higher in the AEAC-treated group, while the expression of the downstream effectors of the low-affinity NGF receptor was significantly lower in the same group. These results suggest that AEAC may contribute to the regulation of NGF expression and secretion in neuronal cells; it is therefore an excellent candidate for further investigation as a therapeutic drug for neurodegenerative diseases.

• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1386-1392
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• 2008

A gene (sll0158) putatively encoding a glycogen branching enzyme (GBE, E.C. 2.4.1.18) was cloned from Synechocystis sp. PCC6803, and the recombinant protein expressed and characterized. The PCR-amplified putative GBE gene was ligated into a pET-21a plasmid vector harboring a T7 promoter, and the recombinant DNA transformed into a host cell, E. coli BL21(DE3). The IPTG-induced enzymes were then extracted and purified using Ni-NTA affinity chromatography. The putative GBE gene was found to be composed of 2,310 nucleotides and encoded 770 amino acids, corresponding to approx. 90.7 kDa, as confirmed by SDS-PAGE and MALDI-TOF-MS analyses. The optimal conditions for GBE activity were investigated by measuring the absorbance change in iodine affinity, and shown to be pH 8.0 and $30^{\circ}C$ in a 50 mM glycine-NaOH buffer. The action pattern of the GBE on amylose, an $\alpha$-(1,4)-linked linear glucan, was analyzed using high-performance anion-exchange chromatography (HPAEC) after isoamylolysis. As a result, the GBE displayed $\alpha$-glucosyl transferring activity by cleaving the $\alpha$-(1,4)-linkages and transferring the cleaved maltoglycosyl moiety to form new $\alpha$-(1,6)-branch linkages. A time-course study of the GBE reaction was carried out with biosynthetic amylose (BSAM; $M_p{\cong}$8,000), and the changes in the branch-chain length distribution were evaluated. When increasing the reaction time up to 48 h, the weight- and number-average DP ($DP_w$ and $DP_n$) decreased from 19.6 to 8.7 and from 17.6 to 7.8, respectively. The molecular size ($M_p$, peak $M_w{\cong}2.45-2.75{\times}10^5$) of the GBE-reacted product from BSAM reached the size of amylose (AM) in botanical starch, yet the product was highly soluble and stable in water, unlike AM molecules. Thus, GBE-generated products can provide new food and non-food applications, owing to their unique physical properties.

• Journal of Dairy Science and Biotechnology
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• v.15 no.1
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• pp.1-9
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• 1997

This study was conducted to efficiently separate the Ig from Korean native cattle colostrum and to utilize them as an immunogen for the production of antibodies aginst rabbit. The results obtained were as follows : 1. About 84% of Ig G could be separated from Korean native cattle colostrum by·gel filtration using Superose 12 column on HPLC. The separation profile of Korean native cattle colostral immunoglobulin was similar that of Holstein colostral Ig. 2. Separation of Korean native cattle colostral Ig by anion exchange chromatography using Mono Q column on HPLC was poor resolution chromatographic pattern. 3. Hi-Trap Protein G column showed better results than the Protein A Sepharose CL-4B column in the Ig G binding capacity from Korean native cattle colostral Ig. 4. Protein G Sepharose Fast Flow system resulted in higher Ig g binding capacity as the industrial size scale-up approach. 5. Sufficient titer reaction of antibody to Korean native cattle colostral Ig G was confirmed by ELISA.

• Korean Journal of Environmental Agriculture
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• v.28 no.3
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• pp.274-280
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• 2009

In order to elucidate the adsorption mechanism of the herbicide, bentazon and its metabolites on soil, their adsorption patterns on soil and six adsorbents were investigated with Freundlich, Langmuir and linear isotherm. Freundlich constants ($K_f$) and maximum adsorption amount($Q^0$) of bentazon on soil was 0.55 and 3.97. Kd and Koc values of it were 0.18 and 18. The all of metabolites used except deisopropylbentazon amounts sorbed on the soil were much higher than bentazon. The most of 8-hydroxybentazon was adsorbed on soil showing $K_f$ = 316.6, $Q^0$ = 3,488, Kd = 29.7 and Koc = 2,970. Bentazon, deisopropylbentazon and 8-hydroxybentazon were shown high affinity to anion exchange regardless of pH and $NH_2$ in low pH range. Reversed phase $C_{18}$ resulted in 100% retention of N-methylbentazon regardless of pH and other metabolites were retained below 40%. The AIBA was strongly adsorbed in silica gel, COOH and cation exchange phase but poor retention was on anion exchange sorbent. 2-Aminobenzoic acid showed an amphipathic nature which had high affinity for COOH and cation exchange phase at pH 7.0 as well as $NH_2$ and anion exchange sorbent at pH 3.0.

• Korean Journal of Food Science and Technology
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• v.16 no.2
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• pp.211-217
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• 1984

To study affinity of proteolytic enzymes to soy proteins, the physicochemical and functional properties of enzymatically modified protein products, kinetic parameters and degree of hydrolysis were measured using trypsin, alcalase (serine type protease) and pronase. Bacterial alcalase and pronase showed much greater affinity to soy protein than animal intestinal trypsin. This effect was very significant when unheated soy isolate was used as a substrate. Specific activities of these enzymes decreased with the increment of substrate concentration (over 2.0%, w/v) when heat denatured soy protein was used as a substrate. However, the decrease in specific activity was negligible at substrate concentrations lower than 2.0%. Polyacrylamide gel electrophoretic results showed that the pattern of 2S protein band changed distinctly in alcalase hydrolysis as compared with those of trypsin and pronase. Protein solubilities of alcalase and pronase hydrolyzates increased by 25-30%, at their pI (pH 5.0) over the control. Virtually no change was observed in solubility by trypsin hydrolysis. Heat coagulability and calcium-tolerance of the protein increased by enzymatic hydrolysis. No clear tendency, however, was observed for emulsion properties, foam expansion and the amount of free -SH groups. The enzyme treatment considerably decreased foam stability.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.7-8
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• 2010

The surface conductive layer (SCL) of chemical vapor deposition (CVD) diamonds has attracting much interest. However, neither photoemission electron microscopic (PEEM) nor micro-spectroscopic (PEEMS) information is available so far. Since SCL retains in an ultra-high vacuum (UHV) condition, PEEM or PEEMS study will give an insight of SCL, which is the subject of the present study. The sample was made on a Ib-type HTHP diamond (001) substrate by non-doping CVD growthin a DC-plasma deposition chamber. The SCL properties of the sample in air were; a few tens K/Sq. in sheet resistance, ${\sim}180\;cm^2/vs$ in Hall mobility, ${\sim}2{\times}10^{12}/cm^2$ in carrier concentration. The root-square-mean surface roughness (Rq) of the sample was ~0.2nm as checked by AFM. A $2{\times}1$ LEED pattern and a sheet resistance of several hundreds K/Sq. in UHV were checked in a UHV chamber with an in-situ resist-meter [1]. The sample was then installed in a commercial PEEM/S apparatus (Omicron FOCUS IS-PEEM) which was composed of electro-static-lens optics together with an electron energy-analyzer. The presence of SCL was regularly monitored by measuring resistance between two electrodes (colloidal graphite) pasted on the two ends of sample surface. Figure 1 shows two PEEM images of a same area of the sample; a) is excited with a Hg-lamp and b) with a Xe-lamp. The maximum photon energy of the Hg-lamp is ~4.9 eV which is smaller that the band gap energy ($E_G=5.5\;eV$) of diamond and the maximum photon energy of the Xe-lamp is ~6.2 eV which is larger than $E_G$. The image that appear with the Hg-lamp can be due to photo-excitation to unoccupied states of the hydrogen-terminated negative electron affinity (NEA) diamond surface [2]. Secondary electron energy distribution of the white background of Figs.1a) and b) indeed shows that the whole surface is NEA except a large black dot on the upper center. However, Figs.1a) and 1b) show several features that are qualitatively different from each other. Some of the differences are the followings: the two main dark lines A and B in Fig.1b) are not at all obvious and the white lines B and C in Fig.1b) appear to be dark lines in Fig.1a). A PEEMS analysis of secondary electron energy distribution showed that all of the features A-D have negative electron affinity with marginal differences among them. These differences can be attributed to differences in the details of energy band bending underneath the surface present in SCL [3].

• Animal Bioscience
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• v.34 no.10
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• pp.1579-1589
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• 2021

Objective: This study was conducted to generate single stranded DNA oligonucleotides with selective affinity to bovine spermatozoa, assess its binding potential and explore its potential utility in trapping spermatozoa from suspensions. Methods: A combinatorial library of 94 mer long oligonucleotide was used for systematic evolution of ligands by exponential enrichment (SELEX) with bovine spermatozoa. The amplicons from sixth and seventh rounds of SELEX were sequenced, and the reads were clustered employing cluster database at high identity with tolerance (CD-HIT) and FASTAptamer. The enriched nucleotides were predicted for secondary structures by Mfold, motifs by Multiple Em for Motif Elicitation and 5' labelled with biotin/6-FAM to determine the binding potential and binding pattern. Results: We generated 14.1 and 17.7 million reads from sixth and seventh rounds of SELEX respectively to bovine spermatozoa. The CD-HIT clustered 78,098 and 21,196 reads in the top ten clusters and FASTAptamer identified 2,195 and 4,405 unique sequences in the top three clusters from the sixth and seventh rounds, respectively. The identified oligonucleotides formed secondary structures with delta G values between -1.17 to -26.18 kcal/mol indicating varied stability. Confocal imaging with the oligonucleotides from the seventh round revealed different patterns of binding to bovine spermatozoa (fluorescence of the whole head, spot of fluorescence in head and mid- piece and tail). Use of a 5'-biotin tagged oligonucleotide from the sixth round at 100 pmol with 4×10⁶ spermatozoa could trap almost 80% from the suspension. Conclusion: The binding patterns and ability of the identified oligonucleotides confirms successful optimization of the SELEX process and generation of aptamers to bovine spermatozoa. These oligonucleotides provide a quick approach for selective capture of spermatozoa from complex samples. Future SELEX rounds with X- or Y- enriched sperm suspension will be used to generate oligonucleotides that bind to spermatozoa of a specific sex type.

• Korean journal of food and cookery science
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• v.13 no.4
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• pp.379-383
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• 1997

To study the effect of amylose on the mechanism of gel formation, swelling power, water solubility, the pattern of alkali gelatinization and gel properties were investigated. The swelling power of waxy maize starch (Amioca) was the highest and followed by those of PFP > Amaizo > Amylomaize VII. Amioca showed higher iodine affinity of soluble amylose than other maize starches. The apparent viscosity of alkali gelatinization measured by Brookfield viscometer for Amaizo was 70${\times}$10$^3$cp which was higher than Amylomaize VII. The apparent viscosity of PFP was increased to 70${\times}$10$^3$cp and then decreased. As maize starches were autoclaved at 121$^{\circ}C$ with starch: water ratio (1 : 9) and autoclaving-cooling cycle were up to 4 times, gel properties were measured by rheometer. Amaizo showed the highest value for hardness, adhesiveness, springiness, and cohesiveness, and hardness of gel after two cycles of autoclaving-cooling was the highest. Gel properties seemed to be affected by the chain length of amylose rather than the content of amylose.