• Title/Summary/Keyword: E. coli group

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Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

Causative Organisms of Neonatal Sepsis (신생아 패혈증의 원인 병원체에 대한 조사)

  • Kim, Kyung-Ah;Shin, Son-Moon;Moon, Han-Ku;Park, Young-Hoon
    • Journal of Yeungnam Medical Science
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    • v.16 no.1
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    • pp.60-68
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    • 1999
  • A nationwide survey was conducted to investigate the annual occurrence rate of neonatal sepsis, maternal risk factors in neonatal sepsis, localized infection in neonates, causative organisms in nosocomial infection and the most common causative organism for neonatal sepsis in Korea. Clinical and bacteriological data wele collected from 37 neonatal units to perform retrospective review of the medical records of the newborn infants who were confirmed as having neonatal sepsis and whose blood culture was collected to isolate organisms for one year study period from January to December in 1997. 78,463 neonates were born at 37 hospital in 1997, and 20,869 neonates were admitted to the neonatal units, During this period, 772 episodes of neonatal sepsis were recorded in 517 neonates. The occurrence rate of neonatal sepsis was 0.73%(0~2.95%). Male to female ratio was 1.15:1, and 303 cases(42.1%) were born prematurely. The main pathogens of early onset of sepsis were S. aureus(20%), S. epidermidis(14.4%) and coagulase negative staphylococcus(14. 4%). Gram negative bacilli including Enterobacter spp (7.2%), E. coli(5.1%), Klepstella(4.5%), Pseudomonas(3.7%) and Enterobacter faectum(3.6%) accounted for 24.1% of sepsis. Group B beta-hemolytic streptococcus were isolated only in two cases. Common obstetric factors were PROM(21.1%), difficulty delivery(18.7%), fetal tachycardia(5.3%), chorioamnionitis(4.9%), and maternal fever(4.7%). The main pathogens of late-onset sepsis were S. aureus(22.3%), S. epidermidis(20.4%) and CONS(9.9%). There were 6 cases(1.0%) of Candida sepsis, Frequent focal infections accompanying sepsis were pneumonia(26.1%), urinary tract infection(10.5%), meningitis(8.2%), and arthritis(3.6%), S. epidermidis(22.0%) and s. aureus(21.7%) were also the most common pathogens in 373 nosocomial infection.

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Changes of Quality Characteristics of Spicy Fermented Pork with Atmosphere Packaging during Storage (함기포장한 양념 발효 돈육의 저장중 품질변화)

  • Jin, S.K.;Kim, I.S.;Hah, K.H.;Lyou, H.J.;Park, K.H.;Lee, J.I.
    • Journal of Animal Science and Technology
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    • v.47 no.5
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    • pp.813-824
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    • 2005
  • This study was carried out to evaluate the quality characteristics of the spicy fermented pork with traditional seasonings. The samples, outside muscle of pork ham, were cut to cube($7{\time}10{\time}2$cm3) and five traditional seasonings such as soy sauce(T1), Kimchi sauce(T2), pickled shrimps sauce(T3), onion sauce (T4) were seasoned by the proportions of meat to seasonings(1:1), respectively. The seasoned samples were fermented at $1{\pm}1^{\circ}C$ for 10 days. And then, the spicy fermented meat was atmosphere packaged and stored at $1{\pm}1^{\circ}C$ for up to 28 days. The pH level of fermented pork has been decreased at the end of the storage compared to the early storage days. The pH level of T3 was higher than that of other treatments during the whole storage days. The salinity and saccharinity of treatments were increased during the storage periods in T1, T3 and T4. TBARS in all treatments was significantly high at the later days of the storage compared to the early days of the storage. VBN was increased until the 21 days of storage and decreased rapidly since then. There were no significant differences in WHC among the all treatments. The shear force was increased with storage in all treatments group. The L* values of surface meat showed a tendency of increasing value along the storage days and the a*, b* values showed a tendency of decreasing value. The number of total bacteria and lactic acid bacteria were tended to increase during storage, while E. coli was decreased as storage period extended. In the results of sensory evaluation, T1 had the highest score in overall acceptability.

Comparison of an Automated Most-Probable-Number Technique TEMPO®TVC with Traditional Plating Methods PetrifilmTM for Estimating Populations of Total Aerobic Bacteria with Livestock Products (축산물가공품에서 건조필름법과 TEMPO®TVC 검사법의 총세균수 비교분석)

  • Kim, Young-Jo;Wee, Sung-Hwan;Yoon, Ha-Chung;Heo, Eun-Jeong;Park, Hyun-Jeong;Kim, Ji-Ho;Moon, Jin-San
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.103-107
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    • 2012
  • We compared between an automated most-probable-number technique $TEMPO^{(R)}$TVC and traditional plating methods $Petrifilm^{TM}$ for estimating populations of total aerobic bacteria in various livestock products. 257 samples randomly selected in local retail stores and 87 samples inoculated with $E.$ $coli$ ATCC 25922, $Staphylococcus$ $aureus$ ATCC 12868 were tested in this study. The degree of agreement was estimated according to the CCFRA (Campden and Chorleywood Food Research Association Group) Guideline 29 and the agreement indicates the difference of two kinds methods is lower than 1 log base 10($log_{10}$). The samples of hams, jerky products, ground meat products, milks, ice creams, infant formulas, and egg heat formed products were showed above 95% in the agreement of methods. In contrast, proportion of agreement on meat extract products, cheeses and sausages were 93.1%, 92.1%, 89.1%, respectively. One press ham and five sausages containing spice and seasoning, two pork cutlets containing spice and bread crumbs, two meat extract product and two natural cheeses and one processing cheese with a high fat content, and one ice cream containing chocolate of all samples showed the discrepancy. Our result suggest that $TEMPO^{(R)}$TVC system is efficient to analyses total aerobic bacteria to compare manual method in time-consuming and laborious process except livestock products having limit of detection.

Sanitary Characteristics of Seawater and Sediments in Tongyeong Harbor (통영항의 해수 및 저질의 위생학적 특성)

  • Park, Jun-Yong;Kim, hhhYeong-In;Bae, Ki-Sung;Oh, Kwang-Soo;Choi, Jong-Duck
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.367-375
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    • 2010
  • The bacteriological and physiochemical analysis of sea water and sediments in Tongyeong harbor was conducted to evaluate sanitary conditions. The samples were collected at 8 stations established once a month from June, 2008 to May, 2009. During the study period, the range of temperature was from 6.7 to $25.2^{\circ}C$, transparency ranged from 1.2 to 2.6 m, chemical oxygen demand ranged from 1.90 to 2.92 mg/L, dissolved oxygen ranged from 6.2 to 10.5 mg/L, dissolved nitrogen ranged from 0.052 to 0.098 mg/L, phosphate ranged from 0.044 to 0.065 mg/L, respectively. Seafood, if eaten raw, carries the risk of food poisoning. Seafood poisoning is often cause by pathogenic microorganism originating from fecal contamination, such as Salmonella sp., Shigella sp. and norovirus. Fecal coliforms are an important indicator of fecal contamination. Therefore, data on fecal coliform are very important for evaluating the safety of fisheries in coastal areas. So, we investigated the sanitary indicate bacteria. The coliform group and fecal coliform MPN's of sea water in Tongyeong harbor were ranged from < 1.8~22,000/100 mL (GM 164.9 MPN/100 mL) and < 1.8~7,900 MPN/100 mL (GM 33.7 MPN/100 mL), respectively. Total coliform were detected 97.0% in 96 of samples and 68.9% of total coliforms were fecal coliforms. These results similar to another seawater detection ratio of total coloforms and fecal coliforms. The Vibrios was isolated and identified with VITEK system. Four hundred eighty strains that were obtained from sea water samples in Tongyeong harbor Detection ratio Vibrio alginolyticus, 34.2%, Vibrio parahaemolyticus, 13.8%, Vibrio vulnificus 10.0%, and V. mimicus 12.5% respectively. Vibrio cholerae O1, was not detected. During the study period, the ranges of water content, ignition loss, COD, and acid volatile sulfates in sediments in Tongyeoung harbor were 41.0~57.4%, 7.8~10.5%, 6.51~9.30 mg/g, 0.04~0.09 mg/g, respectively. Heavy metals in sediment of Tongyeoung harbor were Cd, $0.10{\pm}0.05$; Cu, $4.79{\pm}8.20$; As, $1.95{\pm}0.17$; Hg, $0.10{\pm}0.07$; $Cr^{6+}$, $0.34{\pm}0.12$; Zn, $125.33{\pm}16.40$; Ni, $16.43{\pm}1.93$ mg/kg.

RNA Interference of Chitinase Gene in Spodoptera litura (담배거세미나방(Spodoptera litura) Chitinase gene의 RNA interference)

  • Jeon, Mi Jin;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.202-209
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    • 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.

Effect of cooling water and inverse lighting on short chain fatty acid and blood lipid of broiler chickens in closed poultry house during hot weather (혹서기 무창계사에서 육계의 혈액지질 및 짧은 사슬지방산에 관한 역전점등과 냉각수 효과)

  • 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
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    • 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.

Development of integrated microbubble and microfilter system for liquid fertilizer production by removing total coliform and improving reduction of suspended solid in livestock manure (가축분뇨 내 대장균 제거와 부유물질 저감 효율 향상을 통한 추비 생산용 미세기포 부상분리와 마이크로 필터 연계 시스템 개발)

  • Jang, Jae Kyung;Lee, Donggwan;Paek, Yee;Lee, Taeseok;Lim, Ryu Gap;Kim, Taeyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.2
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    • pp.139-147
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    • 2021
  • Livestock manure is used as an organic fertilizer to replace chemical fertilizers after sufficient fermentation in an aerobic bioreactor. On the other hand, liquid manure disposal problems occur repeatedly because soil spraying is restricted during the summer when the crops are growing. To use liquid fertilizer (LF) as an additional nutrient source for crops, it is necessary to reduce the amount of suspended solids (SS) in the liquid fertilizer and secure stability problems against pathogenic microorganisms. This study examined the effects of the simultaneous SS removal and E.coli sterilization in the LF using the microbubble (MB) generator (FeMgO catalyst insertion). The remaining SS were further removed using the integrated microbubble and microfilter system. During the floating process in the MB device, the SS were removed by 57.9%, and the coliform group was not detected (16,200→0 MPN/100 mL). By optimizing the HRT of the integrated system, the removal efficiency of the SS was improved by 92.9% under the 0.1h of HRT condition. After checking the properties of the treated LF, 64.5%, 70.1%, 54.9%, and 51.5% of the TCOD, SCOD, PO4-P, and TN, respectively, were removed. The treated effluent from such an integrated system has a lower SS content than that of the existing LF and does not contain coliforms; therefore, it can be used directly as an additional fertilizer.