• Title/Summary/Keyword: cell homogenates

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Multiple Binding Affinities for Muscarinic Acetylcholine Receptors in Rat Brain (흰쥐 뇌내(腦內)의 무수카린성 콜린 수용체의 이질성(異質性))

  • Lee, Jong-Hwa;El-Fakahany, Esam E.
    • The Korean Journal of Pharmacology
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    • v.23 no.2
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    • pp.101-111
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    • 1987
  • We investigated the binding properties of $(^3H)$ QNB and $(^3H)$ NMS to mAchR to elucidate the characterstics of mAchR in rat brain by using two different preparations (homogemates & intact brain cell aggregates). The binding properties of both ligands demonstrated high affinity and saturability in both experiments, however $(^3H)$ QNB showed a significantly higher maximal binding capacity than tha ot $(^3H)$ NMS 1. In rat brain homogenates; Displacement of both lignands with several mAchR antagonists resulted in competition curves in accoradnce with the law of massaction for QNB, atropine & scopolamine in thie preparation, also a similar profile was found for the quaternary ammonium analogs of atropine & scopolamine (methyl atropine & methylscopolamine) when $(^3H)$ NMS was used to label the receptors in rat brain. But when these hydrophillic antagonists were used to displace $(^3H)$ QNB, they showed interaction with high- and low-affinity binding sites in brain homogenates. Pirenzepine, the nonclassical mAchR antagonist, was able to displace both ligands from binding sites in this preparation. 2. In intact rat brain cell aggregates; Intact bain cell aggregates were used to elucidate the binding characteristics of $(^3H)$ NMS to mAchR in rat. The magnitude of binding of this ligand was related linearly to the amount of cell protein in the binding assay with a high ratio of total to nonspecific binding. mAchR antagonists displaced specific $(^3H)$ NMS binding according to the law of mass-action, while it was possible to resolve displacement curves using mAchR agonist into high-& low-affinity component. 3. Our results indicate that more hydrophilic receptor ligand $(^3H)$ QNB, displacement experiments in both tissues demonstrated that the lipid solubility of a particulr mAchR ligand might play an important role in determining its profile of binding to the mAchR, and the concentrations of mAchR in rat brain are both on the cell surface (membrane-bound receptor) and in the intracelluar membrane (intermembrane-bound receptor). 4. The results are discussed in terms of the usefulness of dissociated intact rat brain cells in studying mAchR in central nervous system.

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Enzyme-linked immunosorbent assay (ELISA) for the detection of RVS (Retrovirus of Salmonid) (ELISA법을 이용한 연어과 어류의 RVS 검출(Retrovirus of Salmonid) 검출)

  • Oh, Myung-Joo;Yoshimizu, Mamoru
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.169-176
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    • 1996
  • An indirect double antibody enzyme-linked immunosorbent assay (ELISA) was developed for rapid detection of a new virus isolated from abnormally swimming salmonid fish, RVS (Retrovirus of salmonid). Results using brain tissue homogenates, and infected cell cultures are described. The sensitivity of the methods is $10^{2.6}$ $TCID_{50}/100{\mu}l$ of the examined cell culture fluid. The specificity was confirmed by the ELISA inhibition test and virological examinations. Viral antigen could be detected in artificially infected fish tissue homogenates. The assay will allow the diagnosis of RVS-infected fish within a day.

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Changes of Chemical Components During the Storage of Fresh Red Pepper Homogenates (파쇄(破碎) 생(生)고추의 밀봉(密封) 저장중(貯藏中) 품질(品質) 성분(成分)의 변화(變化))

  • Lee, Gye Hee;Oh, Man Jin
    • Korean Journal of Agricultural Science
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    • v.13 no.1
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    • pp.130-138
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    • 1986
  • In order to obtain the basic data for development of raw pepper homogenates as instant spice, effects of preservatives, packaging materials, storage temperature and period on chemical components of raw pepper homogenates were investigated during storage after sealing up. The results are as follows: 1. Raw pepper homogenates added 15% sodium chloride was effective prominently than raw pepper homogenates on residual contents of acidity, capsanthin and capsaicin. 2. P. V. D. C film was effective than P. E film in sealing of raw pepper homogenates. 3. Decomposition of capsanthin was exceeded at high temperature during the sealed storage of raw pepper homogenates and decomposition of capsaicin was accelerated at initial stage of storage and also it was decreased prominently by adding of sodium chloride. 4. Decomposition of vitamin C during the storage was exceeded at high temperature and it was prominently suppressed by adding of sodium chloride. 5. Increase of total viable cell count and lactic acid bacteia count was prominently suppressed by adding of sodium chloride and its difference for storage temperature was disregarded.

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Application of Modified Stokes Expression to Model the Behavior of Expanded Beds with Feed Streams Containing E. coli Homogenates

  • Chae, Young-Rea;Yoon, Yeo-Joon;Ryu, Keun-Garp
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.323-325
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    • 2004
  • To model the behavior of expanded beds with aqueous feed streams containing different amounts of glycerol, we previously developed a modified Stokes expression that correlates the terminal settling velocity of a particle of a solution with the properties of the particle (particle diameter and density) and the solution (density and viscosity). Two empirical parameters, the effective diameter of the poly-disperse resins for protein adsorption and an exponent of non-unity for $(\rho_{P}-\rho)/\mu$ term, are introduced in the modified Stokes expression. We applied the same type of the modified Stokes expression in combination with the Richardson-Zaki correlation to the published results [1], and found that the expansions of the beds with feed streams containing different amounts of E. coli homogenates can also be successfully described.

The Growth of Vibrio vulnificus in Meat Homogenates of Fish and Shellfish (어패육에서의 Vibrio vulnificus의 증식에 관한 연구)

  • KIM Young-Man;HUR Sung-Ho;CHANG Bong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.2
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    • pp.80-84
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    • 1988
  • The change of cell counts of Vibrio vulnificus in meat homogenates of fish and shellfish by the storage time and temperature was examined to get basic information for precautionary steps against septicemia from slices of raw fish (sashimi). Therefore, we inoculated raw and cooked meat homogenates of fish and shellfish with Vibrio vulificus M-8 (isolated from shellfish ) and stored them at $-20^{\circ}C,\;4^{\circ}C\;and\;30^{\circ}C$ for 72 hours. Vibrio vulnificus M-8 was not detected in 32 hours when it was frozen and stored at $-20^{\circ}C$ after inoculating them into phosphate buffer solution at concentration of $10^5\;cell/ml$, while the existance of Vibrio vulnificus was identified after 72 hours of storage at the same temperature in case of inoculation into the meat homogenate of yellow tail. The cell count of Vibrio vulnificus was decreased as about $20\%$ of initial count after 2 hours storage at $4^{\circ}C$ in phosphate buffer solution with fish and shellfish homgenates. From the experimental results it was recognized that Vibrio vulnificus was labile to the cold stress. In comparison to the growth of growth of Vibrio M-8 at $30^{\circ}C$ in the raw and cooked meat of the yellow tail(Seriola guingueradita), snapper(Chrysophrys major), ark shell(Anadra brouhgtonii), and oyster(Crassostrea gigas), the raw meat homogenates were more excellent than the cooked ones though all fish and shellfish meat homogenates were proves to be good for the growth of the microbe.

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The Storage Temperature Effect on Vibrio parahaemolyticus in Fish Homogenates (생선회에 오염원 장염비브리오균에 미치는 저장온도의 영향)

  • KIM Young-Man;LEE Myoung-Suk;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.2
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    • pp.136-140
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    • 1986
  • The change of cell counts of Vibrio parahaemolyticus in fish muscle by the storage time and temperature was examined to get basic informations for precautionary steps against food poisoning of slices of raw fish (sashimi). There fore, we inoculated fish homogenate of oceanic bonito (Katsuwonus pelamis), yellow tail (Seriola quinqueradiata) with Kanagawa positive Vibrio parahaemolyticus and stored it at $30^{\circ}C,\;18^{\circ}C,\;4^{\circ}C\;and\;-20^{\circ}C$ for 24 hours. The number of the Vibrio parahaemolyticus upon fish homogenate stored at $30^{\circ}C\;and\;18^{\circ}C$ decreased for the first two hours and increased thereafter. When the fish homogenates inoculated with Vibrio parahaemolyticus at about $10^3$ per gram were stored at $18^{\circ}C\;and\;30^{\circ}C$ for 10 hours, the cell numbers increased about 10 times and 1,000 times initial cell numbers, respectively. The survival rate of Vibrio parahaemolyticus was about $20\%$, when the inoculated fish homogenates were stored at $-20^{\circ}C$ for 24 hours. Vibrio parahaemolyticus inoculated in fish homogenates was decreased by about $10\%$ of initial cell numbers by the storage at $4^{\circ}C$ for 4 hours and it was decreased by about $50\%$ after 24 hours storage of the samples at the same temperature. The decreasing rate of inoculated Vibrio parahaemolyticus in fresh fish muscle homogenate was higher than that in frozen fish muscle homogenate during the storage time at a refrigerator.

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The Accelerative Effect on Fermentation of Salted and Fermented Anchovy by Homogenates of Sea Tangle, Laminaria japonica Aresschoug (다시마 마쇄물을 이용한 멸치젓갈의 숙성 촉진)

  • BYUN Han-Seok;LEE Tae-Gee;LEE Yong-Woo;PARK Yeung-Beom;KIM Seon-Bong;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.2
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    • pp.127-132
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    • 1994
  • The present study was directed to investigate the accelerative effects of fermentation of salted anchovy on sea tangle homogenates. With the addition of $8\%$ (w/w) sea tangle homogenates(T), there was an increase of amino-N content in both the muscle and juice of salted anchovy during all fermentation periods. It was only in VBN value that there was exhibited the same characteristics as the control batch. Viable cell count in muscle was increased rapidly after 60 days of fermentation, but in juice the content was maximal after 60 days of fermentation. When $8\%$ (w/w) sea tangle homogenates was added, the pH value in muscle and juice were maintained same degree of control until 80 days of fermentation, but showed increase in pH value of muscle and juice more rapidly than the control system after 80 days of fermentation. Then a large percentage of muscle turned to juice after 100 days of fermentation. The degree of fermentation In salted anchovy, when sea tangle homogenates were added, accelerated more than the control batch. Concerning the factors related to the accelerative effects on fermentation of salted anchovy, there was a continuous increase in amino-N content, and it was a sudden change of viable cell count and pH value at a certain point in the fermentation periods.

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The Control of Fermentation Conditions of Salted and Fermented Anchovy by Homogenates of Potato, Solanum tuberosum (감자 마쇄물을 이용한 멸치젓갈의 숙성 조절)

  • BYUN Han-Seok;LEE Tae-Gee;YEO Seang-Gyu;PARK Yeung-Beom;KIM Seon-Bong;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.2
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    • pp.121-126
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    • 1994
  • The present study was designed to investigate effects of the addition of potato homogenates on fermentation control of salted anchovy. With the addition of $8\%$ (w/w) potato homogenates(P), amino-N content and pH were maintained at lower values in the muscle and juice of salted anchovy during all fermentation periods, And high values in external appearance of the salted and fermented anchovy-body during fermentation period were recorded. Consequently, the suppressing effect on the fermentation of salted anchovy of the addition of $8\%$ (w/w) potato homogenates was proved. As for the factors related to the suppressive effects on fermentation, there was no change in amino-N content, and viable cell counts and pH values were maintained.

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Purification of a Thermostable Recombinant Sulfolobus solfataricus Esterase Expressed in a Mesophilic Host

  • Kim, Seong-Hun;Lee, Seon-Bok
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.501-504
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    • 2000
  • The purification of a thermostable esterase expressed in Escherichia coli was investigated using thermoprecipitation of unclarified cell homogenates followed by after applying the heat-treated lysate to phenyl-sepharose column, and elution with detergent. Heat treatment at $70^{cdot}C$ was capable of removing to E. coli proteins. Specially, the thermoprecipitation with 15% polyethylene glycol 8000 can remove host proteins and nucleic acids efficiently. Various detergents were used to recover the esterase, which was strongly bound to phenyl-sepharose resin. Triton X-100, non-ionic detergent, was found to be the most efficient of all tested detergents.

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Metabolism and excretion of novel pulmonary-targeting docetaxel liposome in rabbits

  • Wang, Jie;Zhang, Li;Wang, Lijuan;Liu, Zhonghong;Yu, Yu
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.1
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    • pp.45-54
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    • 2017
  • Our study aims to determine the metabolism and excretion of novel pulmonary-targeting docetaxel liposome (DTX-LP) using the in vitro and in vivo animal experimental models. The metabolism and excretion of DTX-LP and intravenous DTX (DTX-IN) in New Zealand rabbits were determined with ultra-performance liquid chromatography tandem mass spectrometry. We found DTX-LP and DTX-IN were similarly degraded in vitro by liver homogenates and microsomes, but not metabolized by lung homogenates. Ultra-performance liquid chromatography tandem mass spectrometry identified two shared DTX metabolites. The unconfirmed metabolite $M_{un}$ differed structurally from all DTX metabolites identified to date. DTX-LP likewise had a similar in vivo metabolism to DTX-IN. Conversely, DTX-LP showed significantly diminished excretion in rabbit feces or urine, approximately halving the cumulative excretion rates compared to DTX-IN. Liposomal delivery of DTX did not alter the in vitro or in vivo drug metabolism. Delayed excretion of pulmonary-targeting DTX-LP may greatly enhance the therapeutic efficacy and reduce the systemic toxicity in the chemotherapy of non-small cell lung cancer. The identification of $M_{un}$ may further suggest an alternative species-specific metabolic pathway.