• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1995.04a
• /
• pp.82-82
• /
• 1995

4-lBB molecule is expressed on the surface of activated CD4$\^$+/ and CD8$\^$+/ T cells. We generated a panel of anti-4-1 B5 murine mAbs using a fusion protein consisting of the extracellular domain of human 4-1 BB fused to Glutathione S-transferase. The binding activity against cell surface 4-1 BB molecule was assessed by flow cytometry analysis. These studies showed that several anti-4-1 BB mAbs bound to 10-30% of CD4$\^$+/ and CD8$\^$+/T cells in PHA or Con A stimulated PBLs, although these mAbs interacted with only, l-2% of CD4$\^$+/ and CD8$\^$+/ T cells in normal PBLs, indicating the specificity of mAbs to the 4-l BB molecule on activated CD4$\^$+/ and CD8$\^$+/ T cells. Next, we examined the effect of an anti-4-l BB mAb (4B4-1-1) on allogeneic mixed lymphocyte reactions (MLRs). The data indicated that the antibody significantly inhibited the proliferative response at higher concentrations. When tested with several T cell mitogens, the antibody had no stimulatory or inhibitory effects on the mitogen-mediated T cell proliferation. These data suggest that 4-1 BB molecule may play a role in the regulation of antigen-mediated immune response.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.35 no.5
• /
• pp.529-533
• /
• 2002

The peptides inhibiting angiotensin converting enzyme (ACE) were isolated from the hydrolysate of manila clam (Ruditapes philippinamm) proteins prepared with thermolysin. The thermolysin hydrolysate was pretreated with membrane filter (MW cut-off 10,000) to obtain the peptide fraction with ACE inhibition. The crude peptides were applied to a Sephadex LH-20 column and eluted with $30\%$ methanol. The three active fractions (A, B and C) were collected and concentrated, and then applied to a SP-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The four active fractions (A-1, A-2, B-1 and C-1) were collected and concentrated, and then applied to a SuperQ-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The maximum inhibitory activity was observed in the fraction B-1Q showed the IC_{50} values of 0.748 $\mu$g. The abundant amino acids obtained from active fraction B-1Q were leucine, isoleucine, alanine and threonine.

• The Korean Journal of Zoology
• /
• v.37 no.3
• /
• pp.324-330
• /
• 1994

The multicatalvtic 205 proteasome consisting of 12-15 subunits of 22-35 kDa is the catalytic core of the ATP/ubiquitin-dependent 26S protease complex that also is comprised of multiple subunits of 22-110 KDa. In order to determine whether the proteolvtic activities change during muscle development, the enzyme preparations were obtained from 11-, 14- and 17-day old chick embryonic muscle using a BioGel A-1.5m column. The 26S complex preparation from 14- or 17-day old muscle hvdr olvz e d both N -s uccinvl- Le u- Le u -Val-Tvr-7- amido -4- methvlco umarin ( Suc- LLVY- AMC) and ubiquitin-Ivsozvme conjugates about 50% as well as that from 11-day old muscle. In addition, the activity of 20S proteBsome against Suc-LLVY-AMC also decreased by about 20-30%. However, the protein level of 265 complex remained constant during the entire development period, while that of 205 proteasome increased 5- to 6-fold, as analyzed by nondenaturins polyacrvlamide gel elenrophoresis followed by immunoblot analysis using the antibodies raised against the purified enzymes. Thus, the specific activity of 20S proteasome against the peptide must decrease rather dramatically during the muscle development. These results suggest that the development-dependent changes in the proteolytic activities of both 20S proteasome and 26S protease complect from embryonic muscle are due to alterations in the expression of certain subunits in the enzvmes that are responsible for their specific cataIVtic functions but not to overall changes in the enzyme amounts.

• Animal cells and systems
• /
• v.1 no.4
• /
• pp.641-646
• /
• 1997

Lysophosphatidic acid (LPA; 1-acyl-glycerol-3-phosphate) has been known as an intercellular phospholipid messenger with a wide range of biological activities. In this study, the effect of LPA on both the proliferation and differentiation of rat E63 myoblasts has been investigated. In the serum-free Insulin-Transferrin-Selenium (ITS) media, the proliferation of E63 cells was largely restricted. Addition of LPA into the ITS media strongly promoted the cell proliferation and resulted in two to four fold increase of cell number. Furthermore, it appeared to increase the percent fusion in a dose-dependent manner up to 15 ug/ml. The synthesis of myosin heavy chain (MHC) was increased by LPA as well. These results indicate that LPA is able to promote both cell proliferation and differentiation in rat E63 myoblasts. Suramin, known to have uncoupling activity on growth factor-receptor interaction, was tested for antagonistic activity in myoblast proliferation and differentiation. Myoblasts grown in the ITS medium containing LPA were able to proliferate well even in the presence high concentration of suramin whereas myoblast differentiation was completely blocked by 30 ug/ml of suramin. The inhibitory effect of suramin on the myoblast differentiation was completely reversible by removing the suramin. This result indicates that the intracellular signaling pathway of LPA leading to cell proliferation might be distinct from that leading to cell differentiation on E63 myoblasts. Also, the antagonistic effect of suramin suggests that the differentiation activity elicited by LPA might be mediated by a specific G protein-coupled receptor.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.30 no.4
• /
• pp.717-720
• /
• 2001

Chemical compositions and physiological activities of Platycodon grandiflorum roots grown for 4 and 24 years were investigated. Chemical compositions of P. grandiflorum roots grown for 24 years were moisture 82.7%, crude protein 1.6%, crude lipid 2.1% crude ash 0.7%. total sugar 6.0% total dietary fiber 2.3% crude saponin 3.4mg% and ascorbic acid 3.0 mg%. Contents of each components in 24 years old roots were higher than that in 4 years old roots and the most abundant mineral was potassium being 879.9mg/kg. Physiological activities of solvent extract from P. grandiflorum roots grown for 4 and 24 years were high in order of ethanol methanol and water extract. The electron-donating abilities and nitrite-scavenging effect of solvent extract of 24 years were high in order of ethanol methanol and water extract. The electron-donating abilities and nitrite-scavenging effect of solvent extract of 24 years old roots were higher than those of 4 years of old roots in the range of 10~40% respectively.

• Journal of the East Asian Society of Dietary Life
• /
• v.25 no.2
• /
• pp.246-260
• /
• 2015

The purpose of this study was to evaluate the prevalence of obesity, dietary habits, and nutritional status by age among low-income women, using data from the fourth Korea National Health and Nutrition Examination Survey (2007~2009). Subjects were 8,356 women aged 20 and over. The subjects were classified into four groups by age. Dietary data from 24-hr recall methods were used to analyze nutritional status. The prevalence of obesity in the 50~64 years age group was significantly higher than those of the other age groups. Among age groups, malnutrition was the highest in the 65-and-over age group. It appears that women in the 20~29 and 65-and-over age groups were the highest nutritional risk. The percentage of carbohydrates in total energy intake was higher and the percentages of protein and fat were lower in the 65-and-over age group than other groups. Frequency of skipping breakfast was lower in women aged 65-and-over, and moderate physical activity significantly decreased with increasing age. Awareness of dietary guidelines was higher in women aged 30~49 years than other groups, whereas it was lower in those aged 65-and-over years. Adherence to dietary guidelines of 'eating a variety of foods from each food group' was significantly lower in women aged 65-and-over years than those of other groups. However adherence to dietary guidelines of 'eating breakfast everyday with a pleasant mind' was significantly lower in women aged 20~29 years than those of other groups. Therefore, this study shows that low income women have various nutritional problems by age group, and we should support a tailored approach to improve their nutritional status.

• YAKHAK HOEJI
• /
• v.39 no.2
• /
• pp.161-168
• /
• 1995

Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

• KSBB Journal
• /
• v.10 no.3
• /
• pp.343-348
• /
• 1995

Human lipocorin-I(LCI) is a calcium ion-dependent and phospholipid-binding protein which exhibits an anti-inflammatory activity by inhibiting phospholipase A2 activity. In this study, the LCI gene containing its own terminator region was joined to GAL10 promoter-ppL (prepro-leader sequence of mating factor a). An ATG start codon of LCI gene was placed at downstream with KR endoprotease recognition site(Lys-Arg) of ppL. Recombinant S. cerevisiae harboring the LCI expression/secretion vector, pYGLPT5, was aerobicall grown on a liquid YPDG medium al $30^{\circ}C$ for 72hys. The whole cell and culture supernatant were separated after centrifugation, and the expressed LCI was analyzed by SDS-PAGE and western blotting methods. A majority fraction of the expressed LCI was found to be accumulated in the intracellular fraction, resulting in very low secretion efficiency of about 7.4%. About $500mg/\ell$ of LCI was extracellularly produced by the fed-batch culture employing the controlledfeeding of glucose and galactose. The secreted LCI was purified by ultrafiltration and hydroxylapatite column chromatography, and a purity of more than 99% was obtained.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.2
• /
• pp.199-206
• /
• 2012

Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

• Nutritional Sciences
• /
• v.5 no.1
• /
• pp.26-33
• /
• 2002

This study was conducted to provide insight into the management and care of the elderly in nursing homes. Eighty-six elderly subjects over 65 years old (38 male, 48 female) in 3 non-fee-paying nursing homes, located in Daejeon and Chungchong areas, were studied. Subjects'dietary intakes by estimated food intake records, and self-perceived health conditions, were studied using questionnaires from July 21 to August 1, 1997. Twenty-one % of the male subjects and 42% of the female subjects were over 80 years. Their predominant past occupation was farming. While 8.3% of the female and no male residents showed a BMI (Body Mass Index) of 27 and above, about 30% of the subjects were underweight and in poor health status in seeing, joint pains, lumbago and shoulder pains. Regarding overall health status, 72.1% of the subjects considered them to be in poor health, and female and male subjects suffered more difficulties from cloudy eyes, joint pains and lumbago and shoulder pains than any other. Neuralgia was the predominant chronic complaint and followed by hypertension in both sexes. Overall, female subjects felt worse off than the male subjects in terms of their health status, that can be attributed to higher average age of the female subjects compared to the male subjects. The elderly were eating a very low fat (about 15 g : 6% of total calorie) diet with low vitamin A and E. Intakes of calories, protein and iron slightly exceeded RDA. The phosphorus intake was more than double the RDA although calcium intake was close to the RDA. From these results, it seems important to doubling the fat intake of the elderly residents in the form of vegetable fat with the object of raising of vitamin E, antioxidant vitamin, and essential fatty acids for the elderly. It is also recommended that the elderly residents should be given adequate calcium and exercise for bone health.