• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.912-918
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• 2003

Mitotic centromere-associated kinesin (MCAK), which is a novel kinesin with a central motor domain, is believed to playa role in mitotic segregation of chromosome during the M phase of the cell cycle. In the present study, it is shown that a rabbit polyclonal antibody has been produced using the N-terminal region (187 aa) of human MCAK expressed in E. coli as the antigen. To express the N-terminal region in E. coli, the MCAK cDNA fragment encoding N-terminal 187 aa was obtained by PCR and was then inserted into the pET 3d expression vector. Molecular mass of the N-terminal region overexpressed in the presence of IPTG was 23.2 kDa on SDS-PAGE, and the protein was insoluble and mainly localized in the inclusion body that could be easily purified from the other cellular proteins. The N-terminal region was purified by electro-elution from the gel after the inclusion body was resolved on the SDS-PAGE. The antiserum obtained after tertiary immunization with the purified protein specifically recognized HsMCAK when subjected to Western blot analysis, and showed a fluctuation of the protein level during the cell cycle of human Jurkat T cells. Synchronization of the cell-cycle progression required for recovery of cells at a specific stage of the cell cycle was performed by either hydroxyurea or nocadazole, and subsequent release from each blocking at 2, 4, and 7 h. Northern and Western analyses revealed that both mRNA and protein of HsMCAK reached a maximum level in the S phase and declined to a basal level in the G1 phase. These results indicate that a polyclonal antibody raised against the N-terminal region (187 aa) of HsMCAK, overexpressed in E. coli, specifically detects HsMCAK (81 kDa), and it can analyze the differential expression of HsMCAK protein during the cell cycle.

• The Journal of Korean Medicine
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• v.24 no.2
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• pp.1-11
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• 2003

Objectives : The aim of this study was to characterize the effect of Injinchunggan-tang on $TGF-{\beta}1-induced$ hepatic fibrosis. Methods : mRNA and protein expression levels of $TGF-{\beta}1$ in Injinchunggan-tang-treated HepG2 cells were compared to untreated cells using quantitative RT-PCR and ELISA assay, respectively. mRNA expression levels of the TGF-1 pathway genes (TR-1, TR-II, Smad2, Smad3, Smad4, and PAI-1) and fibrosis-associated genes (CTGF, fibronectin, and collagen type 1) were evaluated by quantitative RT-PCR. The effect of Injinchunggan-tang on cell proliferation of T3891 human fibroblast was evaluated using [$^3H$]thymidine incorporation assay. Results : Expression of $TGF-{\beta}1$ mRNA and protein was inhibited by Injinchunggan-tang in a dose- and time-dependent manner. Whereas $TGF-{\beta}1-mediated$ induction of PAI-1 was suppressed by Injinchunggan-tang, expression of the $TGF-{\beta}1$ pathway genes such as TR-1, TR-II, Smad2, Smad3, and Smad4 was not affected by Injinchunggan-tang treatment. Injinchunggan-tang was found to inhibit $TGF-{\beta}1-induced$ cell proliferation of T3891 human fibroblast, and also abrogated $TGF-{\beta}1-mediated$ transcriptional up-regulation of CTGF, fibronectin, and collagen type I. Conclusions : This study strongly suggests that the liver cirrhosis-suppressive activity of Injinchunggan-tang may be derived at least in part from its inhibitory effect on $TGF-{\beta}1$ functions, such as blockade of $TGF-{\beta}1$ stimulation of fibroblast cell proliferation and fibrosis-related gene expression as well as expression of $TGF-{\beta}1$ itself.

• Journal of the East Asian Society of Dietary Life
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• v.22 no.4
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• pp.506-513
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• 2012

This principal objective of this study was to evaluate the sensory properties and flavor compounds of Kimchi prepared with different levels (0.0%, 0.4%, 0.8%, and 1.2%) of Backryeoncho extracts (BE). At high levels of BE, Kimchi showed increased level of crispness and flavor, and also jeotgal odor decreased in the sensory evaluation. Addition of 0.8 % BE resulted in the highest scores for color, taste, and overall acceptance of Kimchi. Therefore, addition of 0.8 % BE appears to be an acceptable approach to enhance the quality of Kimchi without reducing acceptability. As a result of flavor compound analysis, a total of 24 volatile flavor compound, including 11 S-containing compounds, 6 terpenes, 1 acid, 1 ester, 1 alcohol, 2 miscellaneous compounds, 2 thiocyanates, etc., were detected by GC/MS. The major volatile compounds were s-containing compounds and terpene hydrocarbon, and especially terpene of sabinene was newly detected in Kimchi with added BE. Levels of 2-vinyl-[4H]-1,3-dithin derived from garlic flavor as a sulfide-containing compound along with diallyl trisulfide derived from green onion flavor were reduced in Kimchi with added 0.8% BE. Most sulfide-containing compounds were reduced in Kimchi with added BE, whereas most terpenes detected in control Kimchi were not detected.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.726-731
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• 2003

The aim of this research was to identify candidate probiotic lactic bacteria among indigenous dadih lactic isolates. Dadih is an Indonesian traditional fermented milk of West Sumatra which is fermented naturally. Viability of the strain is critical in determining the capacity of lactic bacteria to induce immune stimulation as well as to colonize in the intestinal tract. Therefore, LAB are proposed to exert health promoting or probiotic effects in human, such as inhibition of pathogenic microflora, antimutagenic, and the reduction of cholesterol levels. This manuscript reports in vitro probiotic properties of indigenous dadih lactic bacteria, especially some important colonization factors in GI tract, such as lysozyme, acid and bile tolerance. Bile Salt Hydrolase (BSH) activity, spectrum of bacteriocin, and antimutagenic activity of bacterial cells were also assessed. Twenty dadih lactic isolates were screened further for their tolerance to low pH, at pH 2 and 3 as well as their bile tolerance. There were ten isolates classified as acid and bile acid tolerant, and further screened for lysozyme tolerance, BSH activity. The spectrum of bacteriocin activity of isolates was assayed using cell-free neutralized supernatants by agar spot test against variety of pathogens. Lc. lactis subsp. lactis IS-10285, IS-7386, IS-16183, IS-11857 and IS-29862, L. brevis IS-27560, IS-26958 and IS-23427, Leu.mesen.mesenteroides IS-27526, and L. casei IS-7257 each has good survival rate at low pH values and in the presence of lysozyme, and short lag time in the presence of 0.3 % oxgall. Lc. lactis subsp. lactis IS-11857 and IS-29862 each has high BHS activity, Lc. lactis subsp. lactis IS-10285 and IS-16183 each had a positive spectrum of bacteriocin activity against E. coli 3301 and Lysteria monocytogenes ATCC 19112, while L. brevis IS-26958 has high BHS activity as well as positive spectrum of bacteriocin against E. coli 3301, Lysteria monocytogenes ATCC 19112, and S. aureus IFO 3060. All of the ten dadih lactic strains performed in vitro acid and bile tolerance, indicating a possibility to reach the intestine alive, and display probiotic activities.

• Nutrition Research and Practice
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• v.16 no.3
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• pp.366-378
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• 2022

BACKGROUND/OBJECTIVES: The purpose of this study was to develop a sodium index, which is a tool for estimating and assessing sodium intake easily and quickly, to assist in the prevention of various diseases induced by excess sodium intake in Korean adults. SUBJECTS/METHODS: The 24-h urine collection and dietary behavior surveys were performed on 640 healthy people in 4 regions of South Korea, and an equation for the estimation of 24-h sodium intake was developed. The validity and reliability of the equation were verified with 200 adults. The sodium index was developed by converting the estimated sodium intake using the equation. Finally, the sodium intake status of 1,600 adults was assessed using the sodium index. RESULTS: The equation included sex, age, body mass index, eating habit and dietary behaviors related to sodium intake. In validity test of the equation, the mean bias between sodium intake using 24-h urine analysis and using the equation from the Bland-Altman plots was -1.5 mg/day. The sensitivity and specificity of the equation for estimation of sodium intake were 80.5% and 64.4%, respectively. In the reliability test of the equation, there was no significant difference between the first and second sodium intakes calculated using the equations, and Spearman's correlation coefficient between the 2 sodium intakes was 0.98. Sodium intake can be assessed as 'very moderate' for 75-100 on the sodium index, 'moderate' for 100-150, 'careful' for less than 75 or 150-200, and 'severe' for 250 or more. When sodium intake was assessed using the sodium index in 1,600 subjects, 54.3% and 24.3% of the subjects were assessed to be in the 'careful' and 'severe' categories, respectively. CONCLUSIONS: Using a simple questionnaire, the sodium index can be used to monitor and assess sodium intake status, assisting in nutrition education and counseling in a large population.

• Journal of Food Hygiene and Safety
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• v.13 no.2
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• pp.112-120
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• 1998

This study was performed to investigate the synergistic effect of chitosan and sorbic acid as a new food preservative. So it was performed to investigate inhibitory effect on growh of E. coli 0157:H7, gram negative pathogenic food borne disease bacteria and of S. aureus, gram positive food borne disease bacteria in chitosan, sorbic acid and combination of chitosan and sorbic acid. Minimun Inhibitory Concentration (MIC) of chitosan in E. coli 0157:H7 was 500 ppm at pH 5.0, 250 ppm at pH 5.5, 500 ppm at pH 6.0, and 2000 ppm at pH 6.5, while in Staph. aureus 31.25 ppm at pH 5.0 and 62. 5 ppm at more than pH 5.5. also, MIC of sorbic acid in E. coli 0157:H7 was 500 ppm at pH 5.0, 1500 ppm at pH 5.5, and 2000 ppm at more than pH 6.0, while in Staph. aureus 1500 ppm at pH 5.0 and more than 2000 ppm at more than pH 5.5. Due to the effect of pH in E. coli 0157:H7, MIC of combined chitosan and sorbic acid was 500 ppm of chitosan with 500 ppm of sorbic acid at pH 6.5, but 250 ppm of chitosan with 31.3 ppm of sorbic acid at pH 5.0. In Staph. aureus, there was great effect of chitosan, but neither effect of pH nor sorbic acid. When E. coli 0157:H7 were treated with 500 ppm of chitosan with 500 ppm of sorbic acid and 250 ppm of chitosan with 250 ppm of sorbic acid at pH 6.5, they were inhibited. But, they were increased at the initial concentration of bacteria at 1000 ppm of chitosan in 18 hours, at 500 ppm of chitosan in 36 hours. There was no effect of growth inhibition with sorbic acid but great effect with chitosan on Staph. aureus. The correl~tions between MICs of chitosan and sorbic acid in E. coli 0157:H7 accoding to pH were higher than those in Staph. aureus. R values in E. coli 0157:H7 were 0.95 (p<0.01), 0.99 (p<0.01), 0.97 (p<0.01), and 0.99 (p<0.01) at pH 6.5, 6.0, 5.5, and 5.0 respectively. The synergistic effect of chitosan and sorbic acid in E. coli 0157:H7 could be confirmed from the result of this experiment. Therefore, it was expected that the food preservation would increase or maintain by using sorble acid together with chitosan, natural food additive that did no harm to human body.

• Biomolecules & Therapeutics
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• v.22 no.4
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• pp.347-354
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• 2014

Larrea nitida is a plant that belongs to the Zygophyllaceae family and is widely used in South America to treat inflammatory diseases, tumors and menstrual pain. However, its pharmacological activity remains unclear. In this study we evaluated the property of selective estrogen receptor modulator (SERM) of Larrea nitida extracts (LNE) as a phytoestrogen that can mimic, modulate or disrupt the actions of endogenous estrogens, depending on the tissue and relative amount of other SERMs. To investigate the property of SERM of LNE, we performed MCF-7 cell proliferation assays, estrogen response element (ERE)-luciferase reporter gene assay, human estrogen receptor (hER) binding assays and in vivo uterotrophic assay. To gain insight into the active principles, we performed a bioassay-guided analysis of LNE employing solvents of various polarities and using classical column chromatography, which yielded 16 fractions (LNs). LNE showed high binding affinities for $hER{\alpha}$ and $hER{\beta}$ with $IC_{50}$ values of $1.20{\times}10^{-7}$ g/ml and $1.00{\times}10^{-7}$ g/ml, respectively. LNE induced $17{\beta}$-estradiol (E2)-induced MCF-7 cell proliferation, however, it reduced the proliferation in the presence of E2. Furthermore, LNE had an atrophic effect in the uterus of immature rats through reducing the expression level of progesterone receptor (PR) proteins. LN08 and LN10 had more potent affinities for binding on $hER{\alpha}$ and ${\beta}$ than other fractions. Our results indicate that LNE had higher binding affinities for $hER{\beta}$ than $hER{\alpha}$, and showed SERM properties in MCF-7 breast cancer cells and the rat uterus. LNE may be useful for the treatment of estrogen-related conditions, such as female cancers and menopause.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.394-402
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• 2003

Individual and combined supplementation of phosphorus-adequate, wheat-based broiler diets with exogenous phytase and xylanase was evaluated in three experiments. The effects of the enzyme combination in lysine-eficient diets containing wheat and sorghum were more pronounced than those of the individual feed enzymes. The inclusion of phytase plus xylanase improved (p<0.05) weight gains (7.3%) and feed efficiency (7.0%) of broilers (7-28 days post-hatch) and apparent metabolisable energy (AME) by 0.76 MJ/kg DM. Phytase plus xylanase increased (p<0.05) the overall, apparent ileal digestibility of amino acids by 4.5% (0.781 to 0.816); this was greater than the responses to either phytase (3.6%; 0.781 to 0.809) or xylanase (0.7%; 0.781 to 0.784). Absolute increases in amino acid digestibility with the combination exceeded the sum of the individual increases generated by phytase and xylanase for alanine, aspartic acid, glutamic acid, glycine, histidine, isoleucine, phenylalanine, threonine, tyrosine and valine. These synergistic responses may have resulted from phytase and xylanase having complementary modes of action for enhancing amino acid digestibilities and/or facilitating substrate access. The two remaining experiments were almost identical except wheat used in Experiment 2 had a higher phytate concentration and a lower estimated AME content than wheat used in Experiment 3. Individually, phytase and xylanase were generally more effective in Experiment 2, which probably reflects the higher dietary substrate levels present. Phytase plus xylanase increased (p<0.05) gains (15.4%) and feed efficiency (7.0%) of broiler chicks from 4-24 days post-hatch in Experiment 2; whereas, in Experiment 3, the combination increased (p<0.05) growth to a lesser extent (5.6%) and had no effect on feed efficiency. This difference in performance responses appeared to be 'rotein driven'as the combination increased (p<0.05) nitrogen retention in Experiment 2 but not in Experiment 3; whereas phytase plus xylanase significantly increased AME in both experiments. In Experiments 2 and 3 the combined inclusion levels of phytase and xylanase were lower that the individual additions, which demonstrates the benefits of simultaneously including phytase and xylanase in wheat-based poultry diets.

• IMMUNE NETWORK
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• v.23 no.6
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• pp.48.1-48.21
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• 2023

Mesenchymal stromal/stem cells (MSCs) possess immunoregulatory properties and their regulatory functions represent a potential therapy for acute lung injury (ALI). However, uncertainties remain with respect to defining MSCs-derived immunomodulatory pathways. Therefore, this study aimed to investigate the mechanism underlying the enhanced effect of human recombinant bone morphogenic protein-2 (rhBMP-2) primed ES-MSCs (MSC^BMP2) in promoting Tregs in ALI mice. MSC were preconditioned with 100 ng/ml rhBMP-2 for 24 h, and then administrated to mice by intravenous injection after intratracheal injection of 1 mg/kg LPS. Treating MSCs with rhBMP-2 significantly increased cellular proliferation and migration, and cytokines array reveled that cytokines release by MSC^BMP2 were associated with migration and growth. MSC^BMP2 ameliorated LPS induced lung injury and reduced myeloperoxidase activity and permeability in mice exposed to LPS. Levels of inducible nitric oxide synthase were decreased while levels of total glutathione and superoxide dismutase activity were further increased via inhibition of phosphorylated STAT1 in ALI mice treated with MSC^BMP2. MSC^BMP2 treatment increased the protein level of IDO1, indicating an increase in Treg cells, and Foxp3⁺CD25⁺ Treg of CD4⁺ cells were further increased in ALI mice treated with MSC^BMP2. In co-culture assays with MSCs and RAW264.7 cells, the protein level of IDO1 was further induced in MSC^BMP2. Additionally, cytokine release of IL-10 was enhanced while both IL-6 and TNF-α were further inhibited. In conclusion, these findings suggest that MSC^BMP2 has therapeutic potential to reduce massive inflammation of respiratory diseases by promoting Treg cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.3
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• pp.205-215
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• 2002

The purpose of this study was to evaluate the tissue response in various bone grafting materials, especially xenogenous bone materials in vivo, compare of bone formation capacity of various bone grafting materials on rat skull defects and evaluate the effect of Hyaluronic acid on healing of human Demineralized Freezed Dried Bone allogenous graft (DFDBA) materials in rat calvarial defects. 30 Sprague-Dawly rats were divided into 4 groups. $7{\times}7mm$ size bony defect were artificially prepared in the calvaria (both parietal bone) of all 30 rats and follwed group grafting of autogenous bone graft on right side and allogenic DFDBA on left side bone graft (rat DFDB) in 15 control group, but in 15 experimental group, xenograft (human DFDB) on left side, hyaluronic acid treated with xenograft on right side. Sequential sacrifices was performed at 1, 2, 4, 6, 8 weeks of experiment. These specimens were stained with H&E and MT stain, and then histologic analysis under light microscope was carried out. There were inflammatory reaction in all graft material during early stage. Autogenous and Allogenous DFDBA graft group observed inflammatory reaction at 1 week. Xenograft group persistant inflammatory reaction until 4 weeks, but in HA treated xenograft group inflammatory reaction was decreased at 2 weeks. Osteoblastic activity in control group was begun at 2 week, xenograft group was delayed at 6 weeks, however HA treated xenograft group was begun at 4 weeks. At 2 week, mild osteoclastic activity were observed in all xenograft group not in concerned to HA, but there was no difference each group after 4 weeks. There are most activated angiogenesis around graft mateirals in xenograft group at 2 weeks, but in HA treated xenograft group, decreased angiogenesis was observed at same time. Bone formation and bone maturation of xenograft group, there was no difference in HA treatment, was less than control group. Fibrosis around xenograft materials were observed until 6 weeks, there was no difference between xenograft and HA treated groups.