• Journal of Embryo Transfer
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• v.13 no.2
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• pp.139-145
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• 1998

In the present study, effects of interleukin-2 (IL-2), a differentiator and proliferator of T-cells, on nuclear maturation and sperm penetration of bovine oocytes was examined in a serum-free or serum-containing medium. Basic medium was used TCM-199 supplemented with 2.2g / ι sodium bicarbonate, 100 i.u. /rnl penicillin. 100$\mu$g /ml streptomycin, 0.25$\mu$g/ml Fungizone, this medium treated with FCS and IL-2. In experiment 1, we examined the effect of the addition of 0, 1, 5, 10 or 15nM /ml IL-2 to tissue culture medium (TCM-199) on nuclear maturation of oocytes Development of oocytes to the Metaphase II (M II) stage (%) was significantly (P<0.05) higher at 1, 5,10 and 15 nM /ml IL-2(54.2, 73.5, 80.0 and 69.6%, respectively) than at 0 nM /ml IL-2(35.7%). In experiment 2, we examined the effect of the addition of l0nM /ml IL-2 or 5% FCS in oocyte maturation. Nuclear maturation rates were significantly(P<0.05) higher l0nM /ml IL-2(80%) than non-treatment(35.7%) and 5% FCS(63.6%) treatment. On the other hand, there were no significant difference in the proportion of oocytes developed to the 2-cell stage after addition of IL-2 and/or FCS. These results suggest that IL-2 supports nuclear maturation of bovine immature oocytes in vitro. Serum-free maturation system using IL-2 might be useful for evaluation of various factors on oocyte maturation.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.123-128
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• 2002

The present study aimed at determining the effective dose of Folltropin, a follicle timulating hormone (FSH), on superovulation in indigenous cows of Bangladesh. Fifteen regularly cycling 5～7 years old dry cows, weighing 200～250 kg with 2.5～3.0 body condition scores (BCS) were divided into three groups (n＝5). Individual groups were superovulated with 100, 200 or 300 mg of Folltropin per animal. The superovulation treatment was initiated at Day 10 or Day 11 of the estrous cycle (Day 0＝day of estrus). Alfaprostol (6 mg) was injected to each cow 72 h after the initiation of superovulation treatment to induce eestrus. After confirming standing estrus, the cows were inseminated 2～3 times, 12 h apart, depending on the duration of estrus. At Day 6 or Day 7, individual horns of the uterus were flushed with 150～200 $m\ell$ of phosphate buffered saline supplemented with BSA (0.2％), penicillin (100 IU／$m\ell$) and streptomycin (100 $\mu\textrm{g}$／$m\ell$) using a two-way foley catheter. The embryos were concentrated, removing the excess medium through an embryo filter, and identified under a stereomicroscope. The identified embryos were collected, washed four times, evaluated and graded as excellent, good, fair or poor. The excellent, good and fair embryos were considered as transferable quality embryos. The mean (range). numbers of embryos collected vs. transferable quality embryos far 100, 200 and 300 mg of Folltropin were 4.5 (1～10) vs. 3.5 (1～8); 2.5 (1～4) vs. 1 (0～2) and 0.0 (0～0) vs. 0.0 (0～0), respectively, Folltropin at a dose of 100 or 200 mg produced suitable ovarian stimulation for superovulation in indigenous zebu cows of Bangladesh. A dose of 300 mg or more Folltropin consistently caused preovulatory corpora lutea formation in the ovaries and resulted in zero embryo recovery.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.63-69
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• 2010

The effect of Acibenzolar-S-methyl (ASM) and Rahnella aquatilis Ra39 against apple fire blight disease caused by Erwinia amylovora were tested as a possible alternative to streptomycin. In vitro studies, no inhibition effect against the pathogen was found when ASM was tested. Under greenhouse conditions, application of R. aquatilis Ra39 with the highly susceptible M26 rootstock resulted in a marked disease suppression. Application of ASM and strain Ra39 caused a high decrease of the disease, 82% and 58% respectively; this was correlated with a reduction of the growth of the pathogen within host plants up to 64% and 49.5% respectively. Further studies in the field under artificial infection condition during full bloom revealed that application of ASM and R. aquatilis Ra39 with Gala variety resulted in a control effect up to 21 and 29% respectively. In physiological studies, enhanced activities of PR-proteins (chitinase and $\beta$-1, 3-glucanase) were detected, which are well known as biochemical markers for systemic acquired resistance. Application of ASM to apple shoots caused the highest chitinase activity followed by strain Ra39. The enzyme activity was increased after 2, 4 and 6 days from application. In addition, ASM-treatment caused the higher $\beta$-1, 3-glucanase activity than strain Ra39. Maximum enzyme activity was recorded after 6 days from application and then decreased after 8 and 10 days from application.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.254-254
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• 1996

Eosinophils are known to be important effector cells in pathogenesis of asthma. The elucidation of mechanism by which eosinophil survival is regulated in vivo at sites of inflammation is critical tn our understanding of asthma pathogenesis. The maintenance of these cells at site of inflammation depends upon tile balance between its tendency to undergo apoptosis and tile local eosinophil-viability enhancing activity, Qualitative and quantative phenotypic differences have been observed between bronchoalveolar lavage (BAL) and peripheral blood (PB) eosinophils (EOS). We hypothesize that BAL EOS Possess altered functional feature compared to PB EOS. BAL and PB EOS were obtained from ragweed allergic asthmatics after segmental antigen challenge (SAC) at 24 hour or one week, and purified over percoll and CDl6 negative selection. Cells were cultured in duplicate in RPMI, 15% FCS and 1% penicillin/streptomycin without exogenous cytokines. Eosinophil purity and viability was >92%. BAL. EOS viability was 69${\pm}$4.4% versus 39${\pm}$1.6% for PB EOS (p<0.005) at 48 hour time point, and this difference was maintained through day 5 (32${\pm}$7.6% vs. 3.0${\pm}$ 1.4%, p<0.05), Among BAL EOS, those harvested one week after SAC appeared to have an prolonged survival compared to those harvested at 24 hour. Coculture of BAL and PB EOS resulted in significant viability enhancement than expecteed. Direct neutralization of GM-CSF activity, not IL-3 and EL-5, markedly decreased tile survival of BAL EOS in culture, and abrogated tile viability enhancing activity of their culture supernatants in a dose dependent manner. We conclude that BAL EOS activated in vivo possess enhanced viability compared to PB EOS. Mixing and neutralization experiments suggest a role for autocrine production of GM-CSF.

• Journal of Microbiology and Biotechnology
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• v.20 no.6
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• pp.1042-1052
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• 2010

Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4% of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2%), sulfonamides (57.4%), streptomycin (53.1%), ampicillin (29.7%), nalidixic acid (27.6%), kanamycin (23.4%), chloramphenicol (21.2%), and trimethoprim (19.1%). Resistance towards cephalosporins was noted for cephalothin (27.6%), cephradine (21.2%), amoxicillin clavulanic acid (17.0%), and cephalexin (17.0%). Resistance genes, $bla_{TEM}$, strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirtythree strains (70.2%) carried class 1 integrons, which were grouped in 9 different profiles. DNA sequencing identified sat, aadA, pse-1, and dfrA genes in variable regions on class 1 integrons. Thirty-five strains (74.4%) were subtyped to 22 different plasmid profiles, each with 1-6 plasmids (2.0 to 95 kb). PFGE subtyped the 47 strains into 39 profiles. In conclusion, high rates of multidrug resistance were found among the Malaysian Salmonella Typhimurium strains. The emergence of multidrug-resistant Salmonella Typhimurium to cephalosporin antibiotics was also observed. The strains were very diverse and no persistent clone was observed. The emergence of MDR Salmonella Typhimurium is a worldwide problem, and this report provides information for the better understanding of the prevalence and epidemiology of MDR S. Typhimurium in Malaysia.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.32-32
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• 2002

본 연구는 인공수정용 액상정액을 생산하는 돼지 AI 센터의 종모돈으로부터 채취한 정액내 세균 감염정도를 조사하고 감염율이 높은 세균에 대한 항생제 감수성을 조사하기 위하여 시도되었다. 3개 AI 센터의 원정액내 세균수 (cfu × 10²/㎖)는 각각 8.2±28.8(1 -100), 18.2± 20.0(5-48) 및 33.1±62.l(4-173)로서 평균 23.8±38.l 이었고 AI 센터간, 개체간에 변이 가 컸다. 감염 세 균의 특성은 간균 74%, Gram stain(＋)균 60%, catalase 생산 (＋)균 100% 및 oxidase activity (＋) 균 98%였으며 센터간에 다소 차이가 있었다. 정액샘플내 감연빈도가 높은 세균은 Bacillus sp(조사시료의 75.0%), Pseudomonas sp(67.9%), Proteus sp(53.8%), Staplhylococcus sp(53.6%), E. coli(l5.4%), Klebsiella sp(15.4%), Enterobacter sp(7.7%) 순이었으며 전체 감염세균 종류중 이들 세균의 비율은 각각 25.6%, 20.9%, 16.3%, 18.6%, 4.7% 및 2.3%였다. 액상정액에서 보존 2일과 6일의 세균수 (cfu×10²/㎖)는 2.4± 2.7과 44.0±44.6이었다. 항생제 감수성은 Corynebacterium sp의 경우는 8종 항생제 중 3종 (Streptomycin, polymyxin B, erythromycin)에서 저항성을 나타냈다.(농림기술개발사업 연구결과의 일부)

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.87-93
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• 1982

Two cases of pulmonary disease in a 54 year-old female and a 70 year-old male patient due to Mycobacterium avium-intracellulare complex(MAIC) and a case of pulmonary infection ina 69 year-old male patient due to M. fortuitum(MF) were found recently in this institute. All three patients had a long history of anti-tuberculous chemotherapy because they were initially diagnosed as pulmonary tuberculosis. A 70 year-old male patient infected with MAIC had an unsuccessful chemotherapy history of isoniazid(INH), para-aminosalicylic acid(PAS) and streptomycin(SM) with an incomplete, temporary, symptomatic improvement, for three years since 1964 when he was first diagnosed as pulmonary tuberculosis on physical examination. A 54 year-old female patient infected with MAIC also had an unsuccessful chemotherapy history with the various anti-tuberculous drugs since 1958. Both patients discharged large number of MAIC in their sputum specimens for at least more than one year, but no M. tuberculosis at all. A 69 year-old male patient infected with MF was diagnosed as moderately advanced pulmonary tuberculsis in 1977. Combined chemotherapy with INH+PAS+pyrazinamide(PZA) improved his clinical symptoms, however, his chest radiograph was deteriorated again in 1980 one year after he stopped therapy. Therefore he started chemotherapy again with INH+ethionamide(TH)+cycloserine(CS) but no improvement was noticed. MF was cultured from his sputum in August 1981 and he continuously discharged the same bacilli until last examination of January 1982. Whether all three patients were initially !infected with nontuberculous mycobacteria or complicated with predisposing tuberculosis was not clear because there were no reliable bacteriological examination records.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1276-1284
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• 2006

Listeria monocytogenes is a well-known high-risk foodborne pathogen that grows at refrigeration temperature and is responsible for outbreaks of listeriosis. We report here the incidence of L. monocytogenes in fresh chicken carcasses and present genetic diversity of L. monocytogenes isolates. In this study, 25 g of chicken carcasses from markets in Korea were examined according to the FDA method, and presumptive isolates were confirmed by multiplex PCR assay. L. monocytogenes isolates were analyzed by Pulsed-Field Gel Electrophoresis using restriction enzymes, ApaI and AscI, to obtain strain-specific DNA fragments profiles. Antimicrobial resistance of L. monocytogenes strains against generally used antibiotics (Penicillin G, Kanamycin, Tetracycline, Vancomycin, Cephalothin, Rifampicin, Erythromycin, Ampicillin, Gentamicin, Streptomycin, and Chloramphenicol) were analyzed by NCCLS protocols to examine the presence of antimicrobial resistance in natural L. monocytogenes. Of a total 274 chicken samples, 81 samples (29.6%) were positive for L. monocytogenes. Listeria innocua (50.1%), Listeria welshimeri (6.9%), and Listeria grayi (11.3%) were also detected. PFGE analysis, using restriction enzymes ApaI and AscI, showed 27 pulsotypes of L. monocytogenes. Antimicrobial resistance analysis confirmed the existence of antimicrobial resistance for penicillin G and tetracycline in isolated L. monocytogenes strains.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1061-1065
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• 2001

The antibacterial effects of water extracts of Scutellariate Radix (a dried root of Scutellaria baicalensis GEORGI) and its major flavonoid components, Baicalin and Baicalein, on Salmonella typhimurium, a representative enteric pathogen, were studied. Through a Kriby-Bauer disc analysis, the growth-inhibition activity of Scutellariae Radix against. S. typhimurium was found to be compatible with commercial antibiotics, such as ampicillin, chloramphenicol, and streptomycin. In contrast, the growth of a nonpathogenic E. coli strain was unaffercted by Scutellariae Radix. To examine the effect of polyphosphate kinase (ppk), a putative virulence factor, on the antibacterial activity of Scutellariae Radix, the growth profile of a ppk mutant of S. typhimurium was investigated in a tryptic soy broth containing different concentrations of water extracts of Scutellariae Radix. The ppk mutant was able to grow in 6 mg/ml of water extracts of Scutellariae Radix, whereas in 6 mg/ml of water extracts of Scutellariae Radix, whereas the wild-type could not, implying that the inactivation of ppk made S. typhimurium more resistant to the antibacterial activity of Scutellariae Radix. No enhanced resistance was observed in a ppk mutant of S. typhimurium complemented with a ppk expression vector. The attenuation of the virulence by ppk inactivation was also observed in a virulence assay using BLAB/c mice. Neither Baicalin nor Baicalein exhibited any growth-inhibition activity against S. typhimurium. The water extracts of Scutellariae Radix stimulated the transcription of ppk, especially in the early growth-stage of S. typhimurium.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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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.