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Simultaneous Purification of Enterotoxin A and C by Fast Protein Liquid Chromatography (FPLC에 의한 Staphylococcal Enterotoxin A와 C의 동시분리)

  • Lee, Jung-Hee;Kim, Jong-Bae;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.856-861
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    • 1988
  • A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

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Changes in the Properties of Protein during the Fermentation of Salted Shrimp (새우젓 숙성중의 단백질 특성변화에 관한 연구)

  • Kim, Byung-Mook
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.883-889
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    • 1988
  • The salted small shrimps(Acetes japonicus) were fermented for 3 months at room temperature. During the period of fermentation, the changes of shrimp protein properties were determined. The extractability of soluble protein was slightly decreased in 1 month fermentation, but thereafter increased. The contents of 10% TCA soluble fraction were gradually increased during 3 month fermentation, and the rate of 10% TCA soluble fraction/total soluble protein was also greatly increased during the period of fermentation. Sephadex G-100 gel filtration pattern was changed after 1 month fermentation, showing the disappearance of low molecular weight protein peaks, the decomposition and the delay of elution time of main shrimp protein peaks. Polyacrylamide gel disc electrophoresis patterns showed the degradation of main protein bands into lots of smaller bands after 1 month fermentation. The contents of total free amino acids were slightly decreased in 1 month fermentation and then gradually increased during the Period of fermentation. The rate of free amino acids/soluble protein was steadily increased during the period of fermentation, but the rate of free amino acids/10% TCA soluble fraction was decreased continually during the period of fermentation. The contents of most free amino acids were increased during the period of fermentation, but those of histidine and arginine were greatly decreased in 1 month fermentation. Ammonia was increased after 1 month fermentation. The pH value of salted shrimp was slowly changed during 3 months of fermentation, showing increase from 7.8 to 8.2.

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Study on the Movement of New Qi-gong "WuQinXi" Exercise for Lumbar Spinal Disease : Based on 10 Mode, 15 Mode, 18 Mode (요추 질환에 대한 신기공 오금희의 동작연구 -10식, 15식, 18식을 중심으로-)

  • You, Kyung Gon;Yeom, Seung Ryong;Lee, Sang Yeong;Kwon, Young Dal;Jeong, Hyun Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.3
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    • pp.280-288
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    • 2013
  • Hua-Tuo(145-208) created five "WuQinXi" exercise by imitating the movements of a tiger, a deer, a bear, a monkey and a bird. The "WuQinXi" exercise, one of the medical Qi-gongs, is an exercise maximizing human's self healing power and has been effective significantly at several modern researches. There are many exercise therapies in western medcine, such as Willamss flexion exercise, Mckenzie's extension exercise, vertebral stabilization exercise and so on. However, there isn't a special exercise therapy which can be applied for medical practice in oriental medicine. So We selected some motions from "WuQinXi" exercise, which are suitable for lumbar spinal disease, and analyzed them. After that, We assorted them by kinds of lumbar spinal disease. First, We selected 22 motions which are related with lumbar movements from 3 type "WuQinXi" exercises ; 10 mode, 15 mode, 18 mode. And then, We classified them according to lumbar movements as flexion, extention, lateral bending and rotation, and also functions as stabilization and rubbing. Next, with these classifications, We assorted those motions by kinds of lumbar spinal disease as HIVD(herniation of intervertebral disc), spinal stenosis, spondylolysis and spondylolisthesis, facet joint syndrome, compression fracture and spondylosis. We expect that trying "WuQinXi" exercise at clinic in this way, the particular exercise therapy of oriental medicine, "WuQinXi" exercise will become more popular. And Oriental medical doctors will be able to teach patients "WuQinXi" exercise's motions easily at clinic, depending on kinds of lumbar spinal disease each patient suffers from. We plan to study more about 20 mode, 30 mode, 40 mode and the effect of "WuQinXi" exercise by comparing patients who do the "WuQinXi" exercise with the patients who do the western medical exercise therapy.

Immunoelectrophoretic analysis of major component proteins In cystic fluid of Taenia solium metacestodes (면역전기영동법에 의한 유구낭미충 낭액의 구성 단백질 분석)

  • Yoon Kong;Seung-Yull Cho;Suk-Il Kim;Shin-Yong Kang
    • Parasites, Hosts and Diseases
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    • v.30 no.3
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    • pp.209-218
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    • 1992
  • When cystic fluid of Taenia solium metacestodes (CF) was filtrated through Sephacryl S-300 Superfine, major proteins were in fractions III add IV Major protein in fraction III was Band C protein of 150 kDa and that in fraction IV was Band N protein (Choi et of., 1990). When CF was electrophoresed in 0.9% agarose gel and reacted with anti-CF rabbit serum (RACF), two main bands, a long outer and a short inner band, were precipitated, together with 8 minor bands. RACF reacted with fraction III forming the long outer band whereas RACF formed the short infer band with fraction IV in immunoelectrophoresis (IEP) The long outer precipitin band of CF fraction III was similar to antigen B in hydatid fluid (HF) of Oriol et at. (1971), while the short inner band of CF fraction IV was similar to HF antigen 5 of Caption et at. (1967) . When HF was reacted with RACF, the short inner band was immunoprecipitated without forming the long outer band. Common antigenicity between CF and HF seemed to exist in fraction IV rather than in fraction III of CF. Patient sera of neurocysticercosis reacted more frequently with fraction III than with fraction IV.

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Genetic Transformation of Chrysanthemum with Cold Regulated Gene (BN115) (저온저항성 유전자를 이용한 국화 형질전환)

  • Han, Soo-Gon;Choi, In-Young;Kang, Chan-Ho;Ko, Bok-Rai;Choi, Joung-Sik;Lee, Wang-Hyu
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.19-25
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    • 2006
  • With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

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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The Effect of LSC/GDC (50 : 50 vol%) Active Layers on Oxygen Transport Properties of LSCF/GDC (20 : 80 vol%) Dual-phase Membrane (LSC/GDC (50 : 50 vol%) 활성층이 LSCF/GDC (20 : 80 vol%) 복합 분리막의 산소투과 거동에 미치는 영향)

  • Cha, Da-Som;Yoo, Chung-Yul;Joo, Jong Hoon;Yu, Ji Haeng;Han, Moon-Hee;Cho, Churl-Hee
    • Membrane Journal
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    • v.24 no.5
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    • pp.367-374
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    • 2014
  • In the present study, disc-type LSCF/GDC (20 : 80 vol%) dual-phase membranes having porous LSC/GDC (50 : 50 vol%) active layers were prepared and effect of active layers on oxygen ion transport behavior was investigated. Introduction of active layers improved drastically oxygen flux due to enhanced electron conductivity and oxygen surface exchange activity. As firing temperature of active layer increased from $900^{\circ}C$ to $1000^{\circ}C$, oxygen flux increased due to improved contact between membrane and active layer or between grains of active layer. The enhanced contact would improve oxygen ion and electron transports from active layer to membrane. Also, as thickness of active layer increased from 10 to $20{\mu}m$, oxygen flux decreased since thick active layer rather prevented oxygen molecules diffusing through the pores. And, STF infiltration improved oxygen flux due to enhanced oxygen reduction reaction rate. The experimental data announces that coating and property control of active layer is an effective method to improve oxygen flux of dual-phase oxygen transport membrane.

Physiological Characteristics and Angiotensin Converting Enzyme Inhibitory Activity of Lactobacillus brevis HLJ59 Isolated from Salted Shrimp (국내 새우젓에서 분리한 Lactobacillus brevis HLJ59의 Angiotensin Converting Enzyme 저해활성 및 생리적 특성)

  • Jeon, Chun-Pyo;Kim, Yun-Hoi;Lee, Jung-Bok;Jo, Min-Sub;Shin, Kee-Sun;Choi, Chung-Sig;Kwon, Gi-Seok
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.9-14
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    • 2010
  • In this study, lactic acid bacteria with high angiotensin converting enzyme inhibitor activity were isolated from Korean fermented food, such as kimchi and salted seafood. The strain HLJ59, isolated from salted shrimp showed the highest angiotensin converting enzyme inhibitor activity in DeMan Rogosa Sharpe broth. Optimum growth temperature of Lactobacillus brevis HLJ59 was at $34^{\circ}C$. Acid treatment at pH 3.0 for 1.5 h decreased cell viability from $9.9{\times}10^8$ CFU/ml to $3.11{\times}10^4$ CFU/ml. The bile extract concentration of 0.3%, 0.5%, and 1.0% in MRS broth did not inhibit the growth of HLJ59. Isolated strain HLJ59 showed more sensibility to amikacin, gentamycin, neomycin, streptomycin, kanamycin, cefmetazole, cephalothin, ampicillin, ticarcillin, sulbactam+ampicillin, amoxicillin+clavulanic acid (AMC), tetracycline, and sulfamethoxazole+trime thoprim (SXT) as compare to other 7 different antibiotics. However, it showed more resistance to cefoxatin, ceftnaxone, penicillin, ciprofloxacin, nalidixic acid, lincomycin, and chloramphenicol.

Optimizing Imaging Conditions in Digital Tomosynthesis for Image-Guided Radiation Therapy (영상유도 방사선 치료를 위한 디지털 단층영상합성법의 촬영조건 최적화에 관한 연구)

  • Youn, Han-Bean;Kim, Jin-Sung;Cho, Min-Kook;Jang, Sun-Young;Song, William Y.;Kim, Ho-Kyung
    • Progress in Medical Physics
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    • v.21 no.3
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    • pp.281-290
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    • 2010
  • Cone-beam digital tomosynthesis (CBDT) has greatly been paid attention in the image-guided radiation therapy because of its attractive advantages such as low patient dose and less motion artifact. Image quality of tomograms is, however, dependent on the imaging conditions such as the scan angle (${\beta}_{scan}$) and the number of projection views. In this paper, we describe the principle of CBDT based on filtered-backprojection technique and investigate the optimization of imaging conditions. As a system performance, we have defined the figure-of-merit with a combination of signal difference-to-noise ratio, artifact spread function and floating-point operations which determine the computational load of image reconstruction procedures. From the measurements of disc phantom, which mimics an impulse signal and thus their analyses, it is concluded that the image quality of tomograms obtained from CBDT is improved as the scan angle is wider than 60 degrees with a larger step scan angle (${\Delta}{\beta}$). As a rule of thumb, the system performance is dependent on $\sqrt{{\Delta}{\beta}}{\times}{\beta}^{2.5}_{scan}$. If the exact weighting factors could be assigned to each image-quality metric, we would find the better quantitative imaging conditions.

Effects of Artemisia capillaris Extracts on Intestinal Microflora In vitro and In vivo (In vitro 및 In vivo에서 인진쑥 추출물이 장내미생물에 미치는 영향)

  • Oh, Mi-Hyun;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1587-1594
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    • 2010
  • This study was designed to investigate the effect of Artemisia capillaris extracts on the intestinal microflora. In agar diffusion method, the solvent fractions of Artemisia capillaris showed growth inhibition against the intestinal microflora. In particular, the chloroform fraction of Artemisia capillaris had strong antibacterial activity against Clostridium perfringens, Clostridium difficile, Eubacterium limosum, and Bacteroides fragilis, but did not show antibacterial activity against Bifidobacterium bifidum and Lactobacillus acidophilus. Most chloroform fraction of Artemisia capillaris inhibitory activities were not reduced by heat treatment or pH variation against C. perfringens, C. difficile, E. limosum, and B. fragilis. MICs of the chloroform fraction were 1.25 mg/mL against C. perfringens, E. limosum and B. fragilis and 2.5 mg/mL against C. difficile. MBCs of chloroform fraction were 5 mg/mL against C. perfringens, E. limosum and 2.5 mg/mL against C. difficile, B. fragilis. The ethyl acetate fraction of Artemisia capillaris showed $3.08{\pm}0.03$ mg/10 mg total polyphenol and $1.91{\pm}0.03$ mg/10 mg total flavonoid contents. In vivo tests were performed to investigate the influence of Artemisia capillaris extract on the intestinal microflora in rats. The results showed the possibilities of utilizing Artemisia capillaris extracts as a functional food component to control intestinal microflora.