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Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Identification and Characterization of Paenibacillus polymyxa DY1 Isolated from Korean Soil with New Antibacterial Activity (새로운 항균활성을 보이는 토양 분리 세균 Paenibacillus polymyxa DY1의 분류와 동정)

  • Shin, Eun-Seok;Lee, Hee-Moo;Lee, Bok-Kwon;Kim, Sung-Hoon;Kwon, Sun-Il;Yoo, Kwan-Hee
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.47-53
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    • 2007
  • The DY1 strain of Gram-positive, rod-shaped bacteria was isolated from the soil sample collected from Daeam mountain, Korea. The culture filtrate of DY1 strain showed a broad spectrum of antimicrobial activity on various pathogenic and food poisoning enteric bacterial species tested in vitro. It showed significant growth-inhibitory effect on Salmonella enterica sp., Shigella sp., pathogenic Escherichia coli, Vibrio cholerae, Vibrio parahemolyticus, and Yersinia enterocolitica. For the identification of the DY1 strain, morphological, biochemical and molecular phylogenetic approaches were performed. The DY1 strain was found to be a member of the genus Paenibacillus on the basis of morphological and biochemical analyses. The 16S rDNA of DY1 showed the highest pairwise identity with Paenibacillus polymyxa with 99.79% (1,413 bp/1,416 bp). The antimicrobial entity from DY1 looked different from preciously reported ones and seems to have a great potential to be further studied as a candidate of new antibiotics to control multi-drug resistant pathogens.

Effects of Cordyceps Militaris Extract on Tumor Immunity

  • Ha, Jae-Won;Yoo, Hwa-Seung;Shin, Jang-Woo;Cho, Jung-Hyo;Lee, Nan-Heon;Yoon, Dam-Hee;Lee, Yeon-Weol;Son, Chang-Gue;Cho, Chong-Kwan
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.12-29
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    • 2006
  • Background and Aims : Even though various strategies for cancer treatment have advanced with the remarkable development of genomic information and technology, it is far from giving relief to cancer patients. Recently there is accumulating evidence that the immune system is closely connected to anti-tumor defense mechanisms in a multistage process. This includes tumorigenesis, invasion, growth and metastasis. Cordyceps Militaris, a well-known oriental herbal medicine, is a parasitic fungus that has been used as an immune enhancing agent for a long period of time. However, little is known about the cancer-related immunomodulatory effects and anti-tumor activities. In the present study, we aimed to investigate the effects of Cordyceps Militaris extract (CME) on immune modulating and anti-tumor activity. Materials and Methods : To elucidate the effects of CME on macrophage and natural killer (NK) cell activity, we analyzed nitric oxide (NO) production, NK cytotoxicity and gene expression of cytokines related with macrophages and NK cell activity. Results and Conclusions : CME activated and promoted macrophage production of NO. It also enhanced gene expression of IL-1 and iNOS in RAW 264.7 cells. CME promoted cytotoxicity of NK cells against YAC-1 cells and enhanced NK cell related gene expression such as IL-1, IL-2, IL-12, iNOS, IFN-${\gamma}$ and TNF-${\alpha}$ in mice splenocytes. It also Promoted protein expression of IL-10, IL-12, IFN-${\gamma}$ and TNF-${\alpha}$ in mice splenocytes and inhibited lung tumor metastasis induced by CT-26 cell line compared with the control group. From these results, it could be concluded that CME is an effective herbal drug for modulating the immune system and anti-cancer treatment by promoting macrophage and NK cell activity.

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Risk factors and clinical characteristics of necrotizing enterocolitis in full-term newborns (만삭아에서의 괴사성 장염의 위험인자와 임상증상)

  • Jeoung, Young Mi;Je, Hyun Gon;Son, Sang Hee
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.489-493
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    • 2006
  • Purpose : The purpose of this study was to determine those factors which could contribute to the development of necrotizing enterocolitis(NEC) in fullterm. Methods : We retrospectively reviewed the medical record of 20 full-terms with NEC(${\geq}$modified Bell's staging criteria IIa) who were admitted to the Neonatal Intensive Care Unit of Il Sin Christian hospital from January 1998 through July 2005, and for each case, the next 2 healthy newborns were matched as controls. Results : Mean gestational age and birth weight in the fullterm with NEC group was 38.42 weeks and 2,915 g; in the healthy fullterm without NEC group, it was 38.61 weeks and 3,148 g. When compared with the control group, NEC infants had a significantly higher frequency of chorioamnionitis, protracted diarrhea. As for Apgar score at 1 min <7, respiratory problem, congenital heart disease. there were no differences in frequency of preeclampsia, maternal diabetes, maternal drug abuse, meconium-stained amniotic fluid, polycythemia or exchange transfusion. Conclusion : Most of these full term infants have a predisposing factor before developing NEC. Our study suggested that NEC in fullterm infants was significantly associated with protracted diarrhea, and congenital heart disease.

Extremely Low Frequency Magnetic Field is an Environmental Stress Factor by Exerting Oxidative Stress

  • Park, Yong-Jin;Park, Won-Joo;Yim, Sung-Hyuk;Yang, Seong-Jun;Sun, Yuan Lu;Jeong, Ji-Hoon;Park, Eon-Sub
    • Biomolecules & Therapeutics
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    • v.15 no.1
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    • pp.58-64
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    • 2007
  • The previous study reported the biological effect of magnetic field exerted by acting on endocrine and anti-oxidant system. The present study aims to study whether ELF-MF (extremely low frequency magnetic field) affects the physiological endocrine systems such as thyroid and whether ELF-MF affects the defense system against oxidative stress when it alters the function of thyroid. Finally, we correlate the effects of MF on oxidative stress, and adrenal and thyroid with an environmental stress factor. We exposed sham or MF to rats for 5 or 25 days. After the exposure, we determined pain sensitivity, level of TSH, $T_3$ and free $T_4$ in plasma. We also assayed in whole brain, lipid peroxidation, the activity of enzymatic anti-oxidant defense including superoxide dismutase(SOD) and glutathione peroxidase (GPx), and non enzymatic defense such as reduced or oxidized glutathione contents. MF induced the hypersensitivity to thermal stimuli with the reduction of latency. $T_3$ and $T_4$ levels were also increased by the exposure of MF. In addition, we observed the rise of MDA level in rat brain by MF although the MF did not change superoxide dismutase and glutathione peroxidase activity. The effect of MF on both reduced and oxidized glutathione results in decrease in reduced or oxidized glutathione in whole brain. In every experiment, there was no significant difference in MF influence between short term (5 days) and long term (25 days) exposure. Taken together, MF exposure affects the thyroid hormonal control in brain. The elevated thyroid hormone acts on brain, leading to hyper-utilization of oxygen. This phenomenon may be correlated with oxidative stress resulting from MF exposure. In conclusion, we suggest that MF exposure may be an environmental stress by exerting oxidative stress.

Relationship of Level of Stress, Life Style, Subjective Symptoms and Clinical Diagnosis in Clients taken Multiphasic Screening Program (종합검진 수검자의 스트레스 수준과 생활습관, 자각증상 및 임상적 진단의 관련성)

  • Park, Jun-Han;Chun, Jin-Ho;Kang, Jang-Mi;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Jeong, Kui-Won;Urm, Sang-Hwa
    • Journal of Preventive Medicine and Public Health
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    • v.31 no.4 s.63
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    • pp.728-739
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    • 1998
  • To improve wellness and quality of life by recognizing the health efforts of stress, the author estimated the relationships between stress, subjective symptoms and clinical diagnosis through a questionnaire and a battery of specified laboratory tests; electrocardiography, blood pressure, cholesterol, aspartate aminotransferase(AST), alanine aminotransferase(ALT), gamma glutamyl transferase$(\gamma-GTP)$, fasting blood sugar, gastro-endoscopy or UGI, abdominal sonography, etc. The data was gathered from 337 clients who were undergoing multiphasic screening program at a University Hospital from January to March 1998. The mean age of subjects was $46.5{\pm}11.2$ years and the mean of body mass index was $24.0{\pm}3.7kg/m^2$. The mean vol of stress was $18.5{\pm}6.0$ expressed as the score out of 40. By general characteristics and lift style among male, mean level of stress was significantly higher in case of lower socioeconomic status, habitual drug use, longer daily working time(>10 hours), no regular exercise, drinkers, irregular meal, skip-ping breakfast(p<0.05). In case of female, that was significantly higher in case of lower education, lowe. socioeconomic status, longer daily working time(>10 hours), no regular exercise, drinkers, smokers, irregular meal, skipping breakfast(p<0.05). Significant correlations were observed between stress and subjective symptoms in all kinds of organ system (p<0.01). Correlation coefficients of stress among male were relatively high with neuro-psychiatric symptom$(\gamma=0.476)$ and cardio-vascular symptom$(\gamma=0.361)$ in order, and correlation coefficients of stress among female was highest with neuro-psychiatric symptom$(\gamma=0.371)$. The prevalence of the diagnosis through the battery of laboratory tests was high in upper gastrointestinal disorders and hypercholesterolemia in order in both sex group. Among male the mean score of stress was significantly high in ulcerative peptic disorder of upper gastrointestine and hepatopathy in order (p<0.05) . Among female that was significantly high in diabetes mellitus. In summary, it is likely that there are associations between stress, subjective symptoms and clinical diagnosis. To promote wellness and quality of life it would be of value that periodic stress evaluation program and stress management including apropriate control of smoking and drinking, regular exercise and meal.

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Mouse Single Oral Dose Toxicity Test of Chongmyung-tang Aqueous Extracts (총명탕(聰明湯) 열수(熱水) 추출물의 마우스 단회 경구투여 독성 실험)

  • Hwang, Ha-Yeon;Jang, Woo-Seok;Baek, Kyung-Min
    • The Journal of Internal Korean Medicine
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    • v.35 no.1
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    • pp.37-49
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    • 2014
  • Objectives & Methods : The objective of this study was to evaluate the single oral dose toxicity of Chongmyung-tang (CMT) in ICR mice. Korean traditional herbal prescription CMT has traditionally been used as a neuroprotective for treatment of learning disability and memory improvement. CMT, lyophilized aqueous extracts (yield=9.7%) were administered to female and male mice with oral dose of 2,000, 1,000 and 500 mg/kg (body weight) according to the recommendation of Korea Food and Drug Administration (KFDA) Guidelines. Animals were monitored for mortality, changes in body weight, clinical signs and gross observation during 14 days after administration upon necropsy; organ weight and histopathology of 14 principle organs were examined. Results : We could not find any CMT extracts treatment related mortalities, clinical signs, changes in body and organ weight, or gross and histopathological observations against 14 principle organs up to 2,000 mg/kg in both female and male mice, except for some accidental sporadic findings which did not show any obvious dose-relations and most of which also demonstrated in both the female and male vehicle control mice in this experiments. Conclusions : Based on the results of this experiment, the 50% lethal dose ($LD_{50}$) and approximate lethal dose (ALD) of CMT extracts after single oral treatment in female and male mice can be considered to be over 2,000 mg/kg, and is likely to be safe in humans.

Anti-Inflammatory Activity of Ethanol Extract of Sargassum miyabei Yendo via Inhibition of NF-κB and MAPK Activation (NF-κB와 MAPKs 활성 저해를 통한 미야베 모자반(Sargassum miyabei Yendo) 에탄올 추출물의 항염증 활성)

  • Kim, Min-Ji;Bae, Nan-Young;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Jang, Mi-Ran;Im, Moo-Hyeog;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.442-451
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    • 2016
  • The aim of this study was to investigate the anti-inflammatory effect of Sargassum miyabei Yendo ethanol extract (SMYEE) using RAW 264.7 cells and croton oil-induced Balb/c mice. SMYEE inhibited the production of pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and $IL-1{\beta}$] and nitric oxide in lipopolysaccharide (LPS)-induced inflammatory response. In addition, SMYEE suppressed the expression of inducible nitric oxide, cyclooxygenase-2, and nuclear factor-kappa B. Further, SMYEE inhibited the expression of mitogen-activated protein kinases (MAPKs), such as extra cellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase. In ear edema test, edema formation in the SMYEE treatment was lower than that in the positive control and was similar to that in the prednisolone treatment group. Photomicrographs of mice ear tissue showed a reduction in dermal thickness and number of infiltrated mast cells. Therefore, our results indicate that SMYEE exerts an anti-inflammatory effect via inhibition of nuclear factor ${NF}-{\kappa}B$ and MAPK activation and can be used as a natural source of anti-inflammatory compounds.

Determination of Ethambutol n Human Plasma by a Validated HPLC Method and Its Application to Single-dose Pharmacokinetics (인체혈장 중 에탐부톨의 HPLC 분석법의 검증 및 단일용량 투여에 의한 약물동태 연구)

  • Gwak, Hey-Sun;Park, Kyung-Ho;Choi, Jun-Shik;Song, Gin-A;Sung, Min-Kyung;Jang, Jung-Ok;Lee, Hwa-Jeong
    • Journal of Pharmaceutical Investigation
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    • v.35 no.2
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    • pp.123-127
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    • 2005
  • An HPLC method was employed for the determination of ethambutol in human plasma. After addition of internal standard (IS, octylamine, $2\;{\mu}g/mL$) and alkalinization of the plasma with 5 M sodium hydroxide, the drug and IS were extracted into the mixture of chloroform and diethyl ether (40:60, v/v). Following a 15-min vortex-mixing and a 10min centrifugation, the organic phase was spiked with $100{\mu}L$ of phenylethylisocyanate $(2000{\mu}g/mL)$ for chemical derivatization, mixed for 5 min and evaporated to dryness under a stream of nitrogen. The residue was reconstituted with $100{\mu}L$ of mobile phase and $20{\mu}L$ was injected into C18 column with a mobile phase consisting of methanol:water (70:30, v/v). The samples were detected utilizing an ultraviolet detector at 200 nm. The method was specific and validated with a limit of $0.15\;{\mu}g/mL$. Intra- and inter-day precision and accuracy were acceptable for all quality control samples including the lower limit of quantification. The applicability of this method was demonstrated by analysis of human plasma after oral administration of a single 1200-mg dose to 20 healthy subjects. From the plasma ethambutol concentration vs. time curves, the mean AUC was $9.61{\pm}1.64\;{\mu}g{\cdot}hr/mL$ and Cmax of $2.68\;{\mu}g/mL$ reached 2.73 hr after administration. The mean biological half-life of ethambutol was $3.46{\pm}1.21$ hr. Based on the results, this simple and validated assay could readily be used in any pharmacokinetic studies using humans.

Bioequivalence Study of Toriem® Tablet to Motilium-M® Tablet (Domperidone Maleate 12.72 mg) Evaluated by Liquid Chromatography/Tandem Mass Spectrometry

  • Ryu, Ju-Hee;Choi, Sang-Jun;Lee, Myung-Jae;Lee, Jin-Sung;Kang, Jong-Min;Tak, Sung-Kwon;Seo, Ji-Hyung;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.39 no.1
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    • pp.65-71
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    • 2009
  • The aim of the present study was to evaluate the bioequivalence of two domperidone maleate tablets, Motilium-$M^{(R)}$ Tablet (Janssen Korea Ltd., reference product) and $Toriem^{(R)}$ Tablet (Daewon Pharm. Co., Ltd., test product). Domperidone was extracted by liquid-liquid extraction using tert-butyl methyl ether and separated in less than 3 min on $C_{18}$ reverse-phase column using an isocratic elution. A tandem mass spectrometer, as detector, was used for quantitative analysis in positive mode by a multiple reaction monitoring mode to monitor the m/z $426.1{\rightarrow}119.1$ and the m/z $837.4{\rightarrow}158.2$ transitions for domperidone and the internal standard (roxithromycin), respectively. Calibration curves, from $0.05{\sim}50$ ng/mL of domperidone, showed correlation coefficients (r) higher than 0.9941. Intra day and inter day precision (C.V. %) for quality control were ranged from 10.04 to 16.09% and from 10.87 to 18.69%, respectively. The lower limit of quantification (LLOQ) of domperidone was 0.05 ng/mL. The method described is precise and sensitive and has been successfully applied to the study of bioequivalence of domperidone in 24 healthy Korean volunteers. Twenty-four healthy male Korean volunteers received a single dose of each medicine ($2{\times}12.72\;mg$ domperidone maleate) in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Plasma concentrations of domperidone were monitored for over a period of 24 hr after the administration. $AUC_{0-t}$ (the area under the plasma concentration-time curve) was calculated by the linear trapezoidal rule. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. The 90% confidence intervals for the log transformed data were within acceptable range of log 0.8 to log 1.25 (e.g., $log\;0.92{\sim}log\;1.05$ for $AUC_{0-t}$, $log\;0.81{\sim}log\;1.05$ for $C_{max}$). The major parameters, $AUC_{0-t}$ and $C_{max}$ met the criteria of KFDA for bioequivalence indicating that $Toriem^{(R)}$ tablet is bioequivalent to Motilium-$M^{(R)}$ tablet.