• The Korean Journal of Pharmacology
• /
• v.31 no.1 s.57
• /
• pp.135-140
• /
• 1995

Glycation occurs by covalent binding between the carbonyl group of monosaccharides and the epsilon amino group of amino acid. It can alter the physiological function of proteins and causes the development of diabetic complications. In this study, the influence of glycation on protein binding of warfarin and dansylsarcosine was studied by equilibrium dialysis which was performed for 3 hours at $37^{\circ}C$ in the water bath. The high glycated albumin which contained $50{\pm}16%$ of glycated albumin bound less than natural albumin which contained $8.5{\pm}5.28%$ of glycated albumin, if drugs concentration were more than the albumin concentration. But only warfarin binding showed a significant difference of 6% (P<0.05) when the molar concentration ratio of warfarin per albumin was 3. In consideration of low therapeutic concentrations, low glycated albumin concentrations in the body, and rapid elimination of excessive free drugs, these small increaes of free warfarin concentrations by glycation of albumin are not considered as risk. factors for drug intoxication for diabetics, if renal functions are intact.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.36 no.5
• /
• pp.360-365
• /
• 2010

Introduction: Mammalian tooth pulp is densely innervated by sensory nerves that are mostly C fibers and A delta fibers. However, there is evidence suggesting that many unmyelinated axons in the pulp are in fact parent meylinated axons. Immunohistochemical staining for neurofilament protein 200 kDa (NFP200) was performed to identify the demyelinated but parent myelinated axons. Materials and Methods: The pulp was removed from healthy premolars and 3rd molars extracted from juveniles and adults undergoing orthodontic treatment, and immunohistochemical staining were applied with NPF200 antibodies, which specifically dye myelinated axons. The specimens underwent an electron microscopy examination with diaminobenzidine (DAB) immunostaining after observation and analysis by fluorescence and confocal laser scanning microscopy. Results: The NPF200 immuno-positive axons in the radicular pulp areas were observed as bundles of many nerve fibers. Many small bundles were formed with fewer axons when firing to the coronal pulp areas and then reachrd a different direction. In the radicular pulp, unmyelinated axons and myelinated axons were present together. However, in the coronal pulp, unmyelinated axons were most common and NFP200 immuno-positive unmyelinated axons with a larger diameter than those in the radicular pulp were observed more frequently. On the other hand, most of the immuno-positive unmyelinated fibers were similar in size to that of typically well-known unmyelinated fibers. Conclusion: Myelinated fibers innervated to the dental pulp maintain their myelins in the radicular portion, but these fibers lost myelins in the coronal portion. After the loss of myelin, the size of the axoplasm also decreased.

• Food Science and Biotechnology
• /
• v.17 no.5
• /
• pp.912-918
• /
• 2008

This study was conducted to identify the differences in proteomic characteristics of 'Agakong', recombinant inbred line, and its parental genotypes 'Eunhakong' (Glycine max) and 'KLG10084' (G. soja). The isoflavone content of 'Agakong' was 3 times higher than that of its parental lines. A combined high-throughput proteomic approach was employed to determine the expression profile and identity of proteins using 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. The overall distribution patterns of proteins are quite similar, but lots of protein spot intensities varied among the genotypes. A total of 41 proteins, representing significant difference in the quantities of protein among the lines, were successfully identified. Among them, more than 50% of the proteins identified were subunits of glycinin and $\beta$-conglycinin, 2 major storage proteins. This study showed that the proteomic analysis could help to define specific changes in protein level and composition, which can occur in the generation of new soybean varieties.

• The Journal of Internal Korean Medicine
• /
• v.35 no.3
• /
• pp.317-331
• /
• 2014

Objectives: The root of Platycodon grandiflorum (PG) has been known to possess a range of pharmacological activities including anti-cancer, anti-inflammatory, and anti-oxidant effects. The present study was designed to investigate whether or not PG-induced cell death was connected with autophagy and apoptosis in NCI-H460 human lung cancer cells. Methods: Effects on the cell viability and apoptotic activity were quantified using MTT assays and flow cytometry analysis, respectively. Protein activation was measured by immunoblotting. Autophagy was measured by LC3 immunofluorescence and immunoblotting. ROS production and loss of mitochondria membrane potential (MMP) were checked with flow cytometry analysis. Results: Following exposure to PG, NCI-H460 cell proliferation decreased simultaneously inducing autophagic vacuoles and up-regulation of microtubule-associated protein 1 light chain 3 and beclin-1 protein expressions. Interestingly, pre-treated with autophagy inhibitors, 3-methyladenin or bafilomycin A1 further triggered reduction of cell viability. PG treatment also induced apoptosis that was related modulation of Bcl-2 family proteins, death receptors and activation of caspases. In addition, PG stimulation clearly enhanced loss of MMP and reactive oxygen species (ROS) generation. Conclusions: Our results suggest that PG elicited both autophagy and apoptosis by increasing loss of MMP and ROS production. PG induced-autophagy may play a cell protective role.

• Journal of Nutrition and Health
• /
• v.28 no.7
• /
• pp.585-594
• /
• 1995

Attempts were made to determine the effects of 5-Korean wild vegetabel consumptions on blood glucose leveles and orgen-energy metabolisms in streptozotocin-induced diabetic rats. The 5-Korean wild vegetables were : Cassia tora L.(C.t), Lycium chinese Mill(L.c), Trichosanthes kirilowii Max(T.k), Polygonatum odoratum var. Pluriflorum Ohwi(P.o) and Arctium lappa L(A.l). Sixty male Spargue-Dawley rats(160-220g) were induced diabetes mellitus by streptozotocin injection into the tail vein and were devided into 6 groups : a diabetic control and 5-experimental groups. All groups of the rats were fed on a AIN-76 diet, and the 5-experimental groups were fed with each wild vegetable (10%) for four weeks. An increased tendency in body weight of all the groups was observed and the tendency was more significant in L.c, T.k. and P.o. groups. The organ weight of liver and kidney were higher in L.c. and A.l. groups and lower in T.k. group which has shown the improvement from diabetes. Plasma glucose levels were markedly decreased from the 1st week in C.t, T.k. and P.o. groups and the tendency has lasted throughout the four weeks experimental period. The consumption of P.o. has decreased plasma cholesterol level while any significant difference was not seen in plasma protein levels from all the experimental groups. The level of plasma triglyceride was decreased in P.o. group and the levels of plasma free fatty acids were also significantly lower in P.o. and T.k. groups. The liver protein levels were significantly higher in P.o. and T.k. groups and these two groups also showed the negative or relatively small amount of urinary glucose excretion. The experimental group of T.k. has revealed the decreased level of muscles protein and the increased level of muscle glycogen. The 5-Korean wild vegetables contained dietary fiber and 9-analyzed minerals comparable to the ordinary use vegetables.

• Korean Journal of Poultry Science
• /
• v.3 no.1
• /
• pp.11-18
• /
• 1976

Metabolizable energy (M.E.) values of 12 U.S. feedstuffs and 10 Korean feed ingredients for poultry were determined both by the total collection method and by the chromic oxide indicator method. It was found that M.E. values of most poultry feedstuffs can be measured accurately by either method. Limitation of feed intake to almost maintenance level(approximately 60% of ad libitum) did not increase or decrease the M.E. value of the feeds. An attempt was made to establish a prediction equation to estimate the M.E. values based on the apparent metabolizability of dry matter (D.M.) in the feedstuffs. The results indicated that linear relationships do exist between D. M. metabolizability and M.E. values of carbohydrate-rich feedstuffs (grains and their by-products) or protein-rich feed ingredients (oil seed meals and animal protein feeds) or lipid-rich feeds (fats and oils) as follows: The prediction equation for carbohydrate-rich feedstuffs was Y = 0.0947x - 3.498 ($r^2\;=\;0.99$, Sy.x = 0.015); for protein-rich feed ingredients. it was Y = 0.1234x - 4.898 ($r^2\;=\;0.99$, Sy.x = 0.027); and for lipid-rich feedstuffs it was Y = 0.0844x + 0.774 ($r^2\;=\;0.99$, Sy.x = 0.032). where x = metabolizability of dry matter of feeds in %, and Y=metabolizable energy values in kcal./g. The errors attached to these estimations were relatively small. Thus these prediction equations may be very useful for estimation of the M.E. values from D.M. apparent metaboiizability of feeds, especially in areas of the world where calorimetry is not possible.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.1
• /
• pp.89-95
• /
• 2007

Improvements in the dissolution of proteins in two-dimensional gel electrophoresis have greatly advanced the ability to analyze the proteomes of microorganisms under a wide variety of physiological conditions. This study examined the effect of various combinations of chaotropic agents, a reducing agent, and a detergent on the dissolution of the Streptomyces peucetius cytosolic proteins. The use of urea alone in a rehydration buffer as a chaotropic agent gave the proteome a higher solubility than any of the urea and thiourea combinations, and produced the highest resolution and clearest background in two-dimensional gel electrophoresis. Two % CHAPS, as a detergent in a rehydration buffer, improved the protein solubility. After examining the effect of several concentrations of reducing agent, 50 mM DTT in a rehydration buffer was found to be an optimal condition for the proteome analysis of Streptomyces. Using this optimized buffer condition, more than 2,000 distinct and differentially expressed soluble proteins could be resolved using two-dimensional gel electrophoresis with a pI ranging from 4-7. Under this optimized condition, 15 novel small proteins with low-level expression, which could not be analyzed under the non-optimized conditions, were identified. Overall, the optimized condition helped produce a better reference gel for Streptomyces peucetius.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.5
• /
• pp.439-446
• /
• 2009

The biosynthesis study of antibiotics saframycin (SFM) in Streptomyces lavendulae and safracin (SAC) in Pseudomonas fluorescens demonstrated that 3-hydroxy-S-methyl-O-methyltyrosine (3hSmOmTyr), a nonproteinogenic amino acid, is the precursor of the tetrahydroisoquinoline molecular core. In the biosynthetic gene cluster of SAC/SFM, sacD/sfmD encodes a protein with high homology to each other but no sequence similarity to other known enzymes; sacF/sfmM2 and sacG/sfmM3 encode methyltransferases for C-methylation and O-methylation; and sacE/sfinF encodes a small protein with significant sequence similarity to the MbtH-like proteins, which are frequently found in the biosynthetic pathways of non ribosomal peptide antibiotics and siderophores. To address their function, the biosynthetic cassette of 3h5mOmTyr was heterologously expressed in S. coelicolor and P. putida, and an in-frame deletion and complementation in trans were carried out. The results revealed that (i) SfmD catalyzes the hydroxylation of aromatic rings; (ii) sacD/sacF/sacG in the SAC gene cluster and sfmD/sfmM2/sfmM3 in the SFM cluster are sufficient for the biosynthesis of 3h5mOmTyr; and (iii) the mbtH-like gene is not required for the biosynthesis of the 3h5mOmTyr precursor.

• The Korean Journal of Pharmacology
• /
• v.8 no.2
• /
• pp.55-61
• /
• 1972

Superprecipitation of actomyosin has been considered to be an in vitro model of the muscle contraction. The superprecipitation and ATPase activity (which supplies the energy for contraction) are influenced by several factors which are the large amount of changes in ionic strength, Mg and ATP concentrations. But those behaviors are found to be promptly influenced by the change in a small range of calcium concentration which can be controlled by the cellular function of muscle physiologically only in the presence of the modullatory proteins, tropomyosin and troponin. In order to elucidate the precise roles of calcium in the muscle contraction and relaxation, the effects of calcium on the actin- myosin interaction was observed in the presence of tropomyosin and troponin using the superprecipitation system. The results are summarized as follows: 1. EGTA (glycol ether diaminetetraacetic acid)prolonged the initiation of the superprecipitation of natural actomyosin. 2. Superprecipitation curve was declined by adding EGTA at the time when tile curve reached the half- maximum. The degree of declining was proportional to the amount of EGTA added. Especially, upon adding 0.25 mM EGTA the curve was lowered to the level before the protein superprecipitated. But addition of EGTA did not affect the curve after attaining the maximum. 3. Superprecipitation of Perry myosin B was not affected by EGTA added both before and during the course of the reaction. 4. Tropomyosin did not change the response of Perry myosin B to EGTA added at any time of the reaction. 5. Troponin also did not change the response of Perry myosin B to EGTA. 6. Both tropomyosin and troponin together rendered the Perry myosin B to obtain the same response as natural actomyosin to EGTA. 7. It was concluded that actin-myosin interaction was influenced by the minute change of calcium concentration only in the presence of both tropomyosin and troponin. We could reproduce the contraction and relaxation of the muscle in vitro under the presence of ATP by changing the calcium concentration.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.36 no.3
• /
• pp.237-242
• /
• 2016

Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.