The objective of this study was to select yogurt starter from Korean traditional fermented foods. The 2 strains (KM24, KM32) among 50 strains of isolated lactic acid bacteria selected as starter based on milk clotting ability, antimicrobial activity against various pathogens, tolerance in artificial gastric and bile juice and growth in 10 % skimmed milk. The strains were identified as Lacobacillus plantarum (KM32) and Pediococcus pentosacesus (KM24) by 16S rRNA gene sequencing. Viable cell number of yogurt fermented with mixed strains (KM24 and KM32) was 9.66 log CFU/mL after fermentation for 48 h and maintained $10^9CFU/mL$ during fermentation for 72 h at $37^{\circ}C$. The pH and titratable acidity of mixed cultured yogurt were 4.25% and 0.83% after fermentation for 48 h at $37^{\circ}C$, respectively. The physico-chemical characteristics of mixed cultured yogurt after fermentation for 48 h were $38.45{\mu}g/mL$ (polyphenol content), 48.57% (DPPH radical scavenging activity) and 465.40 cp (viscosity), respectively. The mixed cultured yogurt maintained $10^9CFU/mL$ of lactic acid bacteria during storage 10 days at $4^{\circ}C$. The viable cell number of yogurt prepared with mixed culture(KM32+KM24) maintained higher and than that of control (L. casei) during storage. These results indicated the potential use of selected strains (KM32+KM24) isolated from kimchi as a yogurt starter with strong acid tolerance and probiotics properties.
Jeon, Mi Jin;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
The Korean Journal of Pesticide Science
/
v.18
no.3
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pp.202-209
/
2014
RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.
Park, Sang-Oh;Park, Byung-Sung;Hwangbo, Jong;Choi, Hee-Chul
Journal of the Korean Applied Science and Technology
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v.31
no.1
/
pp.31-43
/
2014
This experiment evaluated the interaction effect of extreme heat diet(EHD), inverse lighting, and cool water on the growth performance of broiler chickens under extreme heat stress. There were 4 experimental groups (T1: EHD 1, 10:00-19:00 dark, 19:00-10:00 light, cold water $9^{\circ}C$; T2: EHD 2, 10:00-19:00 dark, 19:00-10:00 light, cold water $9^{\circ}C$; T3: EHD 1, 09:00-18:00 dark, 18:00-09:00 light, cold water $14^{\circ}C$; T4: EHD 2, 09:00-18:00 dark, 18:00-09:00 light, cold water $14^{\circ}C$), each group composed of 25 broilers and the experiment was repeated 3 times. EHD 1 contained soybean oil, molasses, methionine and lysine. EHD 2 contained all nutrients of EHD 1 and vitamin C additionally. As a result, T1 and T2 displayed higher body weight increase and diet intake compared to T3 and T4 (p<0.05). The weights of their liver and gizzard were similar but the weights of the thymus and bursa F were higher for T1 and T2 compared to that of T3 and T4 (p<0.05). It was observed that T1 and T2 displayed higher concentrations of blood triglyceride, total cholesterol, HDL-C and blood sugar compared to that of T3 and T4 but LDL-C level was higher for T3 and T4 compared to that of T1 and T2 (p<0.05). T1 and T2 displayed higher levels of immunity substances such as IgG, IgA and IgM compared to T3 and T4 but the blood level of corticosterone displayed to be lower for T1 and T2 compared to T3 and T4 (p<0.05). The T1 and T2 contained a higher amount of fecal lactobacillus compared to that of T3 and T4 but the T3 and T4 contained a higher amount of fecal E. coli, total aerobic bacteria, coliform bacteria compared to that of T1 and T2 (p<0.05). T1 and T2 displayed higher concentrations of cecal acetic acid, propionic acid and total short chain fatty acids compared to T3 and T4 but T3 and T4 displayed higher concentrations of butyric acid, isobutyric acid, valeric acid and isovaleric acid compared to T1 and T2 (p<0.05). These results have been observed that broiler chickens exposed to extreme heat stress with feeding EHD, inverse lighting and cold water would improve blood lipid, and elevate the production of immunity substance, beneficial microorganisms, and short chain fatty acids. This provision would also reduce the blood sugar consumption rate as energy sources and these effects will improve the growth performance of the broilers exposed to extreme heat.
For the effect of water-soluble degraded chitosan on the shelf-life of tofu, sterilized distilled water, 0.5% degraded chitosan, 0.5% fumaric acid and 0.5% lactic acid used as an tofu-immersion solutions were investigated by microbial counts, pH, and turbidity during the periods of storage at $4^{\circ}C$. After 2 weeks storage, total aerobic microbial counts in tap water and sterilized distilled water used as an immersion solution were $3.8\;{\times}\;10^8$ and $1.8\;{\times}\;10^8\;CFU/mL$, respectively. In 0.5% fumaric acid and 0.5% lactic acid immersion solutions, the microbial counts were around $10^7\;CFU/mL$ after 2 weeks while the microbial population in 0.5% water-soluble degraded chitosan were, however, $1.6\;{\times}\;10^5\;CFU/mL$ after 2 weeks and $1.7{\times}10^7\;CFU/mL$ after 3 weeks. The lag phase of initial contaminated microbes in 0.5% degraded chitosan solution was longer than those of other treatments. The addition of 0.5% fumaric acid and 0.5% lactic acid decreased the initial pH to pH 5.0, while those of tap water, sterilized distilled water and 0.5% degraded chitosan stabilized the immersion solution at around pH 7.2. All initial pH values were decreased during storage and then slowly increased as storage time was increased. The turbidities in all treatments were increased during storage, but the addition of 0.5% degraded chitosan showed the lowest change, compared to other treatments, showing that the water-soluble degraded chitosan has a good antimicrobial effect and has a potential use to extend the shelf-life of tofu product.
Journal of the Korean Society of Food Science and Nutrition
/
v.32
no.1
/
pp.35-41
/
2003
The study was conducted to develope a rapid method for the detection of Yersinia enterocolitica in spring water and vegetables via multiplex polymerase chain reaction (PCR) technique using ail, yst, uirF and subgenus-specific Y16S primers. Specificity and sensitivity of multiplex PCR and application of best primers for the detection of Y. enterocolitica from spring water and vegetables were investigeted. Y. enterocolitica ATCC 27729 strains gave 356 bP and 200 bp (Y16S) and 134 bp (yst) bands. but Y. enterocolitica ATCC 9610 and ATCC 23715 strains gave 200 bp and 134 bp bands.In the meanwhile, non-pathogenic Yersinia species, such as Y. frederikseni, Y. inter-media, Y. kristenseni and Y. pseudotuberculosis gave only single 200 bp band, and other bacteria including Escherichia coli O157:H7 ATCC 25392, Shigella dysenteri. Staphylococcu aureus ATCC 25923 and Listeria mo-nocytogenes ATCC 19111 did not show any bands. Among primers, yst and Y16S primer showed the best sensitivity. Seven CFU/mL Y. enterocolitica cells could be detected with yst and Y16S primers and the sensitivity was significantly improved by the further 2nd PCR after 38 cycles of first PCR amplication. Spring water, cabbage and mushroom were inoculated with Y. enterocolitica to determine the sensitivity of multiplex-PCR for the rapid detection of Y. enterocolitica. Multiplex-PCR assay could detect 7 or 70 cells in spring water and vegetables using whole cell lysate with repeating PCR amplication.
Kim, Jae-Yeol;Choi, Hyung-Seok;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Min, Kyung-Up;Kim, Yoo-Young;Shim, Young-Soo;Yoo, Chul-Gyu
Tuberculosis and Respiratory Diseases
/
v.49
no.2
/
pp.217-224
/
2000
Background : It is well known that when macrophages are stimulated with endotoxin, they produce a wide variety of cytokine mediators, including TNF-$\alpha$ and IL-1$\beta$. However, there is an alteration in the macrophages' responsiveness when they are challenged with repeated bouts of endotoxin, termed "endotoxin tolerance" which is regarded as a self-protective phenomenon from continuous stimulation. In this study, endotoxin tolerance in the peripheral blood monocytes of sepsis patients was evaluated. Methods : Fourteen patients with organism-documented sepsis were included. The severity of illness was evaluated by APACHE II score. Peripheral blood monocytes were isolated from the patients and diluted to $1{\times}10^5$ well. After stimulation with endotoxin (LPS of E. coli O114 : B4, 100 ng/ml), they were incubated at $37^{\circ}C$ in 5% $CO_2$ incubator for 24 hours. Supernatant was collected for the measurement of TNF-$\alpha$ and IL-1$\beta$ with ELISA method. Peripheral blood monocytes of seven healthy volunteers were used as control. Results : The APACHE II score (mean$\pm$SD) of the patients at the time of blood sampling was 12.2$\pm$5.7. The primary infection foci were urinary tract infection, pneumonia, subacute bacterial endocarditis, and catheter related infection, etc. The causative organisms were gram negative rods (10 cases), gram positive cocci (6 cases) with two cases of mixed infection. Serum TNF-$\alpha$ could be measured in 4 cases with 29.9$\pm$27.7 pg/ml. Serum IL-1$\beta$was measurable in only one patient. The TNF-$\alpha$ level of supernatant of cultured peripheral blood monocytes was 2,703$\pm$2,066 pg/ml in patients and 2,102$\pm$1914 pg/ml in controls. The IL-1$\beta$level of supernatant was 884$\pm$1,050 pg/ml in patients and 575$\pm$558 pg/ml in controls. There was no difference of TNF-$\alpha$ and IL-1$\beta$ level between patients and controls. Conclusion : We cannot prove the phenomenon of endotoxin tolerance in this study. Future study needs to be focused on the more severe sepsis patients who were taken for sampling earlier. Addition of serum to the culture medium could be an another valuable option for the success of this study.
Inappropriate storage of fresh-cut onions may result in losses of good quality. To understand storage conditions for shelf-life and quality of fresh-cut onions, The effect of packing type and storage temperature on the quality of fresh-cut onions was evaluated. Onions stored at $0^{\circ}C$ for 2 months were peeled off after removing root and shoot parts. Each three peeled onions were packed in a polyethylene film (PE, $50{\mu}m$) or in a polyethylene/polypropylene film (PE/PP, $100{\mu}m$) with vacuum treatment (70 cmHg) and stored at different temperatures (4, and $10^{\circ}C$) for 21 days. The following analyses were examined to evaluate the quality of fresh-cut onions: microbial population, surface color, titratable acidity and pH, respiration rate, and sensory quality. Fresh-cut onions stored at $4^{\circ}C$ showed less aerobic and coliform bacterial population than those stored at $10^{\circ}C$ during observation periods. Fungal populations of fresh-cut onions packed in PE film stored at $10^{\circ}C$ increased significantly after 13 days. E. coli was not detected in all treatments during whole storage periods. Surface colors of fresh-cut onions were not affected by packing type and storage temperature, however, color difference (${\Delta}E$) of fresh-cut onions in PE/PP film stored at $10^{\circ}C$ was significantly higher than those of other treatments. Titratable acidity of fresh-cut onions was not affected by packing type and storage temperature. However, pH of fresh-cut onions packed in PE film stored at $10^{\circ}C$ increased gradually over the whole storage period. Fresh-cut onions packed in PE film showed higher $CO_2$ and less $O_2$ concentrations at $10^{\circ}C$ than those at $4^{\circ}C$. The sensory quality of fresh-cut onions was significantly affected by packing type and storage temperature after 13 days. Particularly, vacuum treatment in PE/PP film showed better sensory quality than that of PE film package at the same storage temperature. It was concluded that vacuum treatment and storage at $4^{\circ}C$ could be effective to prolong the quality of fresh-cut onions up to 21 days.
Park, Dae-jin;Lim, Young-soo;Oh, Jee-young;Koh, Kwang-Wook;Song, Sung-Eun;Jo, Eun-Joo
Journal of agricultural medicine and community health
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v.31
no.2
/
pp.187-208
/
2006
Objectives: This study was carried out to evaluate the risk factors of Oral cleft and to inspect the living environments of the rural areas of Sintang, Indonesia Methods: During 3 to 9 August 2004, A questionnaire survey was done for the risk factors of oral cleft. Case group was composed of 11 oral cleft patients who admitted Missionary Hospital whose mother's bloods were analyzed for anemia and hyperlipidemia. Control group was composed of 56 reproductive rural women recruited from near rural villages. Also we surveyed 4 rural areas of Indonesia with simple water test kits. $x^2-test$ for significant difference was analysed. Results: Drinking water was statistically significant risk factor(p<0.05) of oral cleft. Other factors had no statistical significancy. The kind of drinking water was river-originated water. In rural villages, water sanitation state, even boiled water, was very poor. Although $NO_2-N$, $NO_3-N$ was negative, E. coli-form microorganisms were strongly positive in most samples. Total food intake amount was not enough, and vitamin supplements were also under the need. Conclusions: Drinking the contaminated river-water around pregnancy was supposed to be one of the risk factors of oral cleft in Indonesia. Further study is needed for nitrate and mercury.
Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
Journal of Ginseng Research
/
v.41
no.3
/
pp.257-267
/
2017
Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.
Kim, Kyoung-Ran;Byun, Hae-Jung;Cho, Hyun-Nam;Kim, Jung-Hyun;Yang, Seun-Ah;Jhee, Kwang-Hwan
Journal of Life Science
/
v.21
no.1
/
pp.119-126
/
2011
There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).
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