• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1498-1504
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• 2005

Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.

• 한국미생물·생명공학회지
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• 제30권2호
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• pp.129-134
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• 2002

토종닭 소장으로부터 가축 소화기 병원성 유해세균의 분리를 위하여 무작위 선별법과 중층-도말법을 사용하여 총130주의 유산균을 1차 분리하였다. 1차 분리한 130주를 paper disc법을 사용하여 최종적으로 살모넬라 균, 포도상구균, 대장균에 대하여 저해능이 우수한 7주를 분리하였다. 분리 균주의 동정을 위하여 1차로 API CHL kit를 사용하였고, 동정의 정확성을 높이기 위하여 2차로 16S rRNA sequencing 방법을 사용하였다. 그 결과 BD14 균주는 Pediococcus pentosaceus로 동정되었으며 나머지 6주는 Lactobacillus sp.로 동정되어 모두 안전성 있는 유산균임을 확인하였다. 여러 병원균에 가장 항균력이 큰 유산균은 L. pentosus K34이었으며, 이 유산균은 염산과 담즙산에 대한 내성에서도 가장 우수한 균주로 나타났다. 10종의 항생제에 대한 감수성을 조사한 결과 ciprofloxacin에 대하여 모든 균주가 내성을 보였고, BL 균주를 제외하고 모든 균주가 colistin, streptomycin에 대하여 내성을 보였다. 또한 BD14, BD16, K34 균주가 gentamicin에 대한 내성을 보였다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권2호
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• pp.139-147
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• 2013

Lysolipids such as LPA, S1P and SPC have diverse biological activities including cell proliferation, differentiation, and migration. We investigated signaling pathways of LPA-induced contraction in feline esophageal smooth muscle cells. We used freshly isolated smooth muscle cells and permeabilized cells from cat esophagus to measure the length of cells. Maximal contraction occurred at $10^{-6}M$ and the response peaked at 30s. To identify LPA receptor subtypes in cells, western blot analysis was performed with antibodies to LPA receptor subtypes. LPA1 and LPA3 receptor were detected at 50 kDa and 44 kDa. LPA-induced contraction was almost completely blocked by LPA receptor (1/3) antagonist KI16425. Pertussis toxin (PTX) inhibited the contraction induced by LPA, suggesting that the contraction is mediated by a PTX-sensitive G protein. Phospholipase C (PLC) inhibitors U73122 and neomycin, and protein kinase C (PKC) inhibitor GF109203X also reduced the contraction. The PKC-mediated contraction may be isozyme-specific since only $PKC{\varepsilon}$ antibody inhibited the contraction. MEK inhibitor PD98059 and JNK inhibitor SP600125 blocked the contraction. However, there is no synergistic effect of PKC and MAPK on the LPA-induced contraction. In addition, RhoA inhibitor C3 exoenzyme and ROCK inhibitor Y27632 significantly, but not completely, reduced the contraction. The present study demonstrated that LPA-induced contraction seems to be mediated by LPA receptors (1/3), coupled to PTX-sensitive G protein, resulting in activation of PLC, PKC-${\varepsilon}$ pathway, which subsequently mediates activation of ERK and JNK. The data also suggest that RhoA/ROCK are involved in the LPA-induced contraction.

• 한국임상수의학회지
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• 제20권2호
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• pp.177-184
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• 2003

The present study was conducted to investigate isolation and antimicrobial drug susceptibility of clinical mastitic milking total 610 (897 quaters) dairy cattles from 36 dairy farms in Gyeongnam south area (Cosung, Masan) during the period from March 1999 to August 2002. The results obtained were summerized as follows . 1. Incidence of bacterial infection in four quaters was showed that right anterior quarter was 178(19.8%), right posterior quarter was 292(32.6%), left anterior quarter was 148 (16.2%), and left posterior quarter was 279 (31.1%), respectively. Isolation rate of posterior two quarters was higher 2 times than anterior two quarters. 2. Incidence of double infections of 897 clinical mastitic milk were showed that single infection was 549 (61.2%), double infection was 167(18.6%), triple infection was 9(1%) and no isolation was 172(19.2%). 3. Isolation of infected etiologic bacteria was showed that Streptococci spp., was 267(31%), Staphylococci spp., was 267(41%), S aureus, was 48(5.6%), G(-) bacillus was 126(5.6%), and Corynebactrium spy. was 52(6%), respectively, from total 861 samples. 4. The results of antimicrobial drug susceptibility of all isolates were showed that Streptococci spp., Staphylococci spp., S aureus, (G)(-) bacillus, and Corynebactrium spp. were susceptible to cefuroxime, cefoperazone, amoxicillin, cefazolin, ampicilin, penicillin, gentamicin, erythromycin, cloxacilin, ciprofloxacin, sulfamethoxasole/trimetoprim, teteracyclin, and norfloxacin (> 70%), but some of them were resistant to neomycin, streptomycin colistin, and cephalothin(> 60). 5. The results of drug susceptibility obtained from each farms had different susceptibility, even though, etiological microorganisms were same in each farms.

• 대한수의학회지
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• 제51권1호
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• pp.29-35
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• 2011

Lack of hygiene and puerperal mastitis are common causes of bacterial diseases in nursing neonates. The aim of this study was to isolate microorganisms from milk samples of healthy female Jindo dogs with suckling puppies and to investigate antimicrobial susceptibility against the isolated bacteria. Milk samples were collected from 120 udders of 12 lactating Jindo dogs that were 2~4 years old without any clinical diseases including mastitis. Bacteria were isolated from 64 milk samples (53.3%), either singly (76.6%) or in combination (23.4%). Staphylococcus (S.) spp. was the most common microorganisms (74.7%) isolated from canine milk, followed by Haemophillus spp. (10.9%), Streptococcus spp. (9.6%), Gardnerella spp. (2.4%) and Moraxella spp. (2.4%). The most frequently isolated organism was S. warneri (31.3%). Antimicrobial susceptibility of these bacteria was tested with 17 antimicrobial agents by Kirbyand Bauer standardized disc diffusion method. Results indicated that bacteria isolated from healthy canine milk were mostly susceptible to amoxicillin-clavulanic acid, cephalothin and ceftiofur, but were resistant to erythromycin, neomycin and tetracycline.

• Journal of Chest Surgery
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• 제2권1호
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• pp.41-49
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• 1969

The authors made a clinical study of 80 cases of empyema who were diagnosed and treated at department of chest surgery, St. Mary`s Hospital, Chatholic Medical College, during the period of May.l964 through April.1969 and compared the empyema of infant and children with that of adults. 1. In age and sex ditribution, infant was 6 cases, childhood 22 cases and adult 52 cases. The ratio of male to female was 2.2:1. There`s a little difference in infant-childhood but prominence of males over females in adults was being 3. 3:1, in its ratio. 2. The cardinal symptoms were cough [61.3%], fever [60.0%] and dyspnea [52.8%]. The leukocytosis were observed in 83.7% of all cases, 96.2% of infant-childhood and 76.9% of adults. The hemoglobin level showed subnormal in 82.1% of infant-childhood and in 55.8% of adults. 3. Most frequent lesion to predisposing factor of empyema was pneumonia [43.7%],being prominent in infants children [64.3%] to that of adult 4. The Pathogenic organism by culture in 75 cases of empyema were staphylococuss [48%], streptococuss[9.3%], Gram[-] bacilli [9.3%], Klebsiella[2.7%], pneumococcus[4.0%], E. coli [5.4%] and no growth 21.3% in over all. Among the cases of empyema. staphlocal origin was 62.9% in infant-childfood and 39.6% adults. 5. Staphylococci were most susceptible to erythromycin [86. 1%], Kanamycin [75.0%], albamycin [61.7%] and neomycin [52.8%] but most resistant to penicillin, Chtoramphenicol and terramycin. 6. In the treatment of empyema, of 53 cases were closed thoracotomy drainage and the remainder of cases by open thoracotomy, decortication, thoracoplasty and pleuropneumonectomy. we could attain favourable results by only the closed thoracotomy in infant-childhood, 28 cases. 7. The mortality rate was 6.3% in over all; adult 3 cases, infant and children 2 cases. 3 cases of these, were due to staphylococcal infection.

• 대한의생명과학회지
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• 제12권3호
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• pp.249-254
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• 2006

It has been reported that liver is a very important organ to xenotransplantation. Pig is known to be a most suitable species in transplantation of human organs. However, the physiological function of pig hepatocytes is not clear elucidated. Epidermal growth factor (EGF) is known to be a mitogen in various cell systems. Thus, we examined the effect of EGF on cell proliferation and its related signal cascades in primary cultured pig hepatocytes. EGF stimulates cell proliferation in a dose (>1ng/ml) dependent manner. EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by AG 1478 ($10^{-6}M$, an EGF receptor antagonist) genistein and herbymycin A (tyrosine kinase inhibitors, $10^{-6}M$), suggesting the role of activation and tyrosine phosphorylation of EGF receptor. In addition, EGF-induced increase of $[^3H]-thymidine$ incorporation was prevented by neomycin $(10^{-4}M)$, U73122 $(10^{-5}M)$ (phospholipase C [PLC] inhibitors), staurosporine ($(10^{-8}M)$, or bisindolylmaleimide I $(10^{-6}M)$ (protein kinase C [PKC] inhibitors), suggesting the role of PLC and PKC. Moreover, EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by PD 98059 (a p44/42 mitogen activated protein kinase [MAPK] inhibitor), SB 203580 (a p38 MAPK inhibitor), and SP 600125 (a JNK inhibitor). EGF increased the translocation of PKC from cytosol to membrane fraction and activated p42/44 MAPK, p38 MAPK and JNK. In conclusion, EGF stimulates cell proliferation via PKC and MAPK in cultured pig hepatocytes.

• 한국수정란이식학회지
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• 제28권3호
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• pp.289-295
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• 2013

Xenotransplantation of pig organs into primates results in fatal damage, referred as hyperacute rejection (HAR), and acute humoral xenograft rejection (AHXR), to the organ graft mediated by antibodies pre-existing and newly-producing in primates against their cognate pig antigens. Functional ablation of ${\alpha}1$,3-galactosyltransferase (Gal-T KO) of pig which is an enzyme involved in synthesis of Gala1-3Galb1-4GlcNAc-R antigen is essentially required to prevent HAR. Moreover, additional genetic modification under Gal-T KO background for enforced expression of human complement regulatory proteins which can inhibits complement activation is known to effectively imped HAR and AHXR. In this study, we constructed a membrane cofactor protein (MCP) expression cassette under control of human $EF1{\alpha}$ promoter. This cassette was inserted between homologous recombination regions corresponding to Gal-T locus. Subsequently this vector was introduced into ear skin fibroblasts of female pig by nucleofection. We were able to obtained 40 clones by neomycin selection and 4 clones among them were identified as clones targeted into Gal-T locus of MCP expression cassette by long-range PCR. Real time RT-PCR was shown to down-regulation of Gal-T expression. From these results, we demonstrated human $EF1{\alpha}$ promoter could induce efficient expression of MCP on cell surface of fibroblasts of female pig.

• Journal of Plant Biotechnology
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• 제43권2호
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• pp.240-247
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• 2016

In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

• 대한수의학회지
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• 제44권1호
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• pp.89-98
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• 2004

This study was carried out to investigate the serotype, and antimicrobial susceptibility and analyze the plasmid profile for the 145 isolates of L. monocytogenes isolated from livestock products and these product processing plants in Gyeongnam, Korea. All of L. monocytogenes strains belonged to serotype 1/2b (57.9%), 1/2a (20.0%), 4b (11.4%), 1/2c, 3b, 4c (each 2.9%) and 4d (0.7%). Serotype 1/2b, 1/2a, 4b from each source were found predominantly. Serotype 1/2b was predominantly higher than other serotype, and there was no significant difference between serotypes isolated from livestock products and product processing plants. 4b was major serotype isolated from raw milk and pork, and serotypes isolated from beef, chickens and slaughterhouse were 1/2b and 1/2a. The susceptibility of 145 strains of L. monocytogenes to 14 antibiotics commonly used in veterinary and human therapy was determined by disk diffusion method. All of L. monocytogenes strains were susceptible to amikacin, ampicillin, cephalothin, chloramphenicol, gentamicin, kanamycin, neomycin and penicillin. L. monocytogenes strains had the highest resistance with colistin (100%), oxytetracycline (44.8%), tetracycline (43.4%) followed by erythromycin (2.8%), spectinomycin (1.4%) and streptomycin (0.7%). Tetracycline resistance, and serotype distribution of the isolates from sample sources were significantly different. Resistance to at least one antibiotic was observed in all of them and 7 different resistant profiles were recorded. The most common resistance pattern were CL-OTC-TC (colistin-oxytetracycline-tetracycline) (42.8%). Among all tested isolates, two different plasmid profiles were observed. Of the 97 examined strains, 14 (14.4%) contained either the 8 and 11 kb plasmid or the 11 kb.