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
/
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
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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
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v.32
no.1
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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.
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
/
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.
Kim, Kyoung-Ran;Byun, Hae-Jung;Cho, Hyun-Nam;Kim, Jung-Hyun;Yang, Seun-Ah;Jhee, Kwang-Hwan
Journal of Life Science
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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).
To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.
Journal of the Korea Academia-Industrial cooperation Society
/
v.7
no.6
/
pp.1319-1336
/
2006
The successive round of talks oil Korea-USA Free Trade Agreement (FTA) has continued, and it also has the Intellectual property(IPR) unit. Until now, tile one of most disputing concerns in IPR unit through talks is the limitation of compulsory license of claimed invention. The US is urging to establish a safeguard for IPR, as similar measure of the US, to protecting the profit of the US enterprises through these on-going talks, it is more likely expected to take the offensive about infringement of the patent seriously. Based on the current circumstances, the provision strategy study is needed to obtain Korea inventors the first inventor defense under the US patent law system as well as understand the current Korea's patent law and its revision against that in the US. In patent Law, both nations with first to file system and first to invent system permit a prior user of an invention to continue to use the invention notwithstanding its subsequent patenting by another under being subject to certain qualifications and limitations, even though a patent by a later inventor is granted. Normally, the first inventor defense has been used to compensate the drawbacks of the first to file system. The US patent Law, however, adopting the first to invent system admits the first inventor defense. Therefore, pursuing counteract provision under consideration with Korean patent Law system and research environment along with investigating the reason why the US adopted its patent law system, the scope of right, and the new reform of Act. 2005 of the institute, which promotes the first Korean inventor to possess the defense right of the US, provides certain preparations for Korean companies against the expected offensive from the US ones under the US patent Law system.
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