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Changes of Cell Surface Hydrophobicity of a Serratia marcescens with Cultivation Time and Temperatures (배양온도와 시간에 따른 Serratia marcescens 표면의 소수성 성질변화)

  • 이상열;신용철;권헌영;조무제;강은경
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.227-232
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    • 1990
  • S. marcescens cultured at $30^{\circ}C$ with vigorous shaking was shown to produce red-pigment, prodigiosin, in the senescent phase of growth. Also, it showed many hydrophobic characteristrics, which were tested by the adherence to noncharged surfaces of polystyrene dishes, a typical agent for the binding of hydrophobic cells and molecules. However, when the cell was cultured at $37^{\circ}C$, it no longer produced either red pigment or hydrophobic materials. Therefore, the bacteria cultured at $37^{\circ}C$ was completely washed-out from the polystyrene dishes at the copious washing step with tap water, in contrast to the cell cultured at $30^{\circ}C$ which was sticked onto the polystyrene dishes very tightly. The lipid compositions extracted from the S. marcescens cultured at $30^{\circ}C$ or $37^{\circ}C$ were very different from each other; the phospholipids, glycolipids and unidentified lipids were produced from the cell cultured at $30^{\circ}C$, whereas large amounts of serratamolide, amphipathic compound, were produced from the cell cultured at $37^{\circ}C$. The data suggest that the pronounced cell surface hydrophobicity of the S. marcescens is mediated by a combination of several surface factors that were affected by cultivation time and temperatures.

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Comparison of Major Nutrients in Eels Anguilla japonica Cultured with Different Formula Feeds or at Different Farms

  • Seo, Jae-Sung;Choi, Jae-Hee;Seo, Ji-Hun;Ahn, Tae-Ho;Chong, Won-Seog;Kim, Seung-Hoon;Cho, Hye-Sun;Ahn, Jun-Cheul
    • Fisheries and Aquatic Sciences
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    • v.16 no.2
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    • pp.85-92
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    • 2013
  • To determine the relative importance of two main factors, diet or culture environment, that affect the proximate composition and main nutritive ingredients (vitamin A, vitamin E, cholesterol, fatty acid composition) in cultured eels, we analyzed the composition of eels fed diets of formula feed (FF) produced by four different companies and of eels cultured at five different eel farms that provided only one of the four different FFs. The four commercial eel FFs did not markedly differ in proximate composition or major nutritive compounds, and consequently, these variables did not significantly differ in cultured eels fed the different FFs. The FF imported from Japan was marginally superior to the two domestic commercial FFs and the FF imported from Taiwan in terms of the proximate composition and main nutritive ingredients of both the FF itself and the eels cultured on it. However, proximate composition and main nutritive ingredients significantly differed among eels cultured at the five farms that used a different FF and among eels fed the four different FFs at the same farm. In conclusion, the difference in quality between domestic and Japanese FFs in terms of eel culture was small, whereas physical or chemical environmental differences among farms during eel culture may more strongly affect the proximate composition and levels of the main nutritive ingredients in cultured eels.

Chondrocyte Culture from Epiphyseal Plate and its Morphological Changes in Autologous Implants of Rabbit (토끼 성장판 연골세포 배양과 자가 이식편에서의 형태학적인 변화)

  • 양영철;정해일;최장석
    • Journal of Life Science
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    • v.10 no.4
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    • pp.408-421
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    • 2000
  • We tried to establish the culture method of the chondrocyte isolated from the epiphyseal cartilage and to investigate morphological changes of chondrocyte cultured with enzyme-digested costal cartilage, the perichondrium and experimentally damaged meniscus of rabbit. De novo chondrocyte pellets were prepared from epiphyseal plates by culturing isolated epiphyseal chondrocytes from 4 week. old rabbits. We morphologically assessed the cartilage formation of the chondrocyte culture with enzyme-digested costal carilage, the perichondrial culture, the cultured chondrocytes transplants into experimentally damaged meniscus of rabbits, the perichondrial culture, the cultured chondrocytes transplants into experimentally damaged meniscus of rabbit. In the 24 days, the epiphyseal chondrocytes maintained the typical phenotypes of the partial nodular cell formation. The 30 days cryopreserved chondrocytes showed abnormal and irregular shape. In the type II collagen added culture, the chondrocytes showed expanded rough endoplasmic reticulum and small and large round-like vesicles of processes. In the type IV collagen added culture, the chondrocytes showed large perinuclear vaculoes and abundant well-developed rough endoplasmic reticulum of processes. In the culture with enzyme- digested costal cartilage and the perichondrial culture, the chondrocytes showed a few swelling rough endoplasmic reticulum and vacuoles. The cultured epiphyseal chondrocytes maintained typical phenotype and the chondrocytes were grown faster and maintained more typical phenotype in the type II and IV collagen added culture. The transformed chondrocytes secreted abundant extracellular matrix in the type II collagen added culture, and showed processes in the type IV collagen added culture. The perichondrial chondrocytes were grown faster and their implants were able to transplant. The cultured chondrocytes transplanted into experimentally damaged meniscus were adapted between the meniscus tissues. And the immunocyto-chemical reaction of the type II collagen of the chondrocytes were found to be maintained. The chondrocytes cultured cartilage. The chondrocytes secreted abundantly. The cultured chondrocytes transplanted into experimentally damaged meniscus changed immature cells into enlarged mature cells with extracellular secretion.

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Electrochemical Characteristics of Osteoblast Cultured Ti-Ta Alloy for Dental Implant (골아세포가 배양된 치과 임플란트용 Ti-Ta합금의 전기화학적 특성)

  • Kim, W.G.;Choe, H.C.;Ko, Y.M.
    • Journal of the Korean institute of surface engineering
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    • v.41 no.2
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    • pp.69-75
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    • 2008
  • Electrochemical behaviors of surface modified and MC3T3-E1 cell cultured Ti-30Ta alloys have been investigated using various electrochemical methods. The Ti alloys containing Ta were melted by using a vacuum furnace and then homogenized for 6 hrs at $1000^{\circ}C$. MC3T3-E1 cell culture was performed with MC3T3-E1 mouse osteoblasts for 2 days. The microstructures and corrosion resistance were measured using FE-SEM, XRD, EIS and potentiodynamic test in artificial saliva solution at $36.5{\pm}1^{\circ}C$. Ti-Ta alloy showed the martensite structure of ${\alpha}+{\beta}$ phase and micro-structure was changed from lamellar structure to needle-like structure as Ta content increased. Corrosion resistance increased as Ta content increased. Corrosion resistance of cell cultured Ti-Ta alloy increased predominantly in compared with non cell cultured Ti- Ta alloy due to inhibition of the dissolution of metal ion by covered cell. $R_p$ value of MC3T3-E1 cell cultured Ti-40 Ta alloy showed $1.60{\times}10^6{\Omega}cm^2$ which was higher than those of other Ti alloy. Polarization resistance of cell-cultured Ti-Ta alloy increased in compared with non-cell cultured Ti alloy.

Effects of Gonadotropin-Releasing Hormone on in vitro Gonadotropin Release in Testosterone-Treated Immature Rainbow Trout

  • Kim, Dae-Jung;Kim, Yi-Cheong;Aida, Katsumi
    • Animal cells and systems
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    • v.13 no.4
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    • pp.429-437
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    • 2009
  • The control mechanism of gonadotropin-releasing hormone (GnRH) on gonadotropin (GTH) release was studied using cultured pituitary cell or cultured whole pituitary obtained from Testosterone (T) treated and control immature rainbow trout. The release of FSH was not changed by salmon type GnRH (sGnRH), chiken-II type (cGnRH-II), GnRH analogue ([des-$Gly^{10}D-Ala^6$] GnRH ethylamide) and GnRH antagonist ([Ac-3, 4-dehydro-$Pro^1$, D-p-F-$Phe^2$, D-$Trp^{3,6}$] GnRH) in cultured pituitary cells of T-treated and control fish. Indeed, FSH release was not also altered by sGnRH in cultured whole pituitary. All tested drugs had no effect on the release of LH in both culture systems of control fish. The levels of LH, in contrast, such as the pituitary content, basal release and responsiveness to GnRH were increased by T administration in both culture systems. In addition, the release of LH in response to sGnRH or cGnRH-II induced in a dose-dependent manner from cultured pituitary cells of T-treated fish, but which is not significantly different between in both GnRH at the concentration examined. Indeed, LH release was also increased by sGnRH in cultured whole pituitary of T-treated fish. GnRH antagonist suppressed the release of LH by sGnRH ($10^{-8}\;M$) and GnRH analogue ($10^{-8}\;M$) stimulation in a dose-dependent manner from cultured pituitary cells of T-treated fish, and which were totally inhibited by $10^{-7}\;M$ GnRH antagonist. These results indicate that the sensitivity of pituitary cells to GnRH is elevated probably through the T treatment, and that GnRH is involved in the regulation of LH release. GnRH-stimulated LH release is inhibited by GnRH antagonist in a dose-dependent manner. The effects of gonadal steroids on FSH levels are less clear.

Changes of Physicochemical properties During Storage at an Aquarium Tank of Wild and Cultured Gray mullet (Mugil cephalus) and Red seabream (Pagrus major) (횟집 수조 보관 중에 자연산, 양식산 참돔 및 숭어의 물리·화학적 성분변화)

  • CHO, Young-Je;PARK, Hyun-Kyu;KIM, Seung-Mi;SHIM, Kil-Bo
    • Journal of Fisheries and Marine Sciences Education
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    • v.21 no.3
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    • pp.451-458
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    • 2009
  • The purpose of this study is to analyze the nutrition composition and physiological changes, and to evaluate the food quality of live fish in cultured and wild fishes which have been kept in an aquarium tank. The moisture and lipid content of wild and cultured fishes when kept in an aquarium tank for seven-days storage was found to be lower than those of the initial stage storage(zero day). The breaking strength was also rapidly decreased in all of live fishes tested in this study as the periods of storage extended. The protein content did not differ significantly. However, the content of cortisol, which is a indicator indicating a stress reaction in tissues, was apt to increasing as the periods of storage extended. The cortisol content of wild fishes were higher than those of cultured fishes. On the other hand, the activity of activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which is also a indicator indicating a stress reaction, were not changed in the serum of wild and cultured fish, suggesting the ALT and AST activity dose not directly related with a healthy loss originated from stress. The death ratio of wild fishes were higher than cultured ones due to limited activity and stress during the storage in a aquarium tank.

Localization of Angiotensin II in Korean Bovine Follicles and Its Effects on IVM/IVF of Oocytes (한우 난소 내 Angiotensin II의 분포와 이의 첨가가 체외성숙 및 수정에 미치는 영향에 관한 연구)

  • Quen, J. H.;M. H. Lee;S. K. Kim
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.59-63
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    • 2004
  • 1. The concentrations of Ang. II were 7.20.91 ${\times}$ $10^3$ , 3.80.34 ${\times}$ $10^3$, 3.50.30 ${\times}$ $10^3$, 2.80.22 ${\times}$ $10^3$ pg/ml in bovine follicular fluids from 1∼3 mm, 3∼5 mm, 5∼7 mm and 8∼10 m follicles, respectively. The concentrations of Ang. II decreased in follicular fluids from large follicles. 2. When oocytes were cultured in media containing various concentrations of Ang. II, a higher proportion of oocytes developed to MII stage in medium with 100 ng/ml (79.5%) Ang II compare to that without Ang. II (58.8%). When oocytes from different sizes of follicles were separately cultured in media containing 100 ng/ml Ang. II, maturation rates were higher in oocytes from small and medium follicles those from controls. 3. GSH content in oocytes cultured for 24 hrs in TCM-199 medium containing 10 and 100 ng/ml of Ang. II was also higher than that of oocytes cultured in medium containing 0 or 10 ng/ml Ang. II. When oocytes were cultured in media containing 0, 10, 100, 1,000 ng/ml of Ang. II, the concentrations of GSH were 5.1M, 5.5M, 7.2M, 8.7M, respectively. 4. When oocytes were cultured in media containing various concentrations of 10, 100, 1,000 ng/ml Ang. II, in vitro maturation and developmental rates were 84.0%, 90.0%, 78.0% and 28.0%, 36.0%, 20.0%, respectively. When oocytes were cultured with an addition of Ang. II in media, in vitro maturation rates higher than that of their controls (76.0%).

Inhibition of angiotensin converting enzyme increases PKCβI isoform expression via activation of substance P and bradykinin receptors in cultured astrocytes of mice

  • Jae-Gyun Choi;Sheu-Ran Choi;Dong-Wook Kang;Hyun Jin Shin;Miae Lee;Jungmo Hwang;Hyun-Woo Kim
    • Journal of Veterinary Science
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    • v.24 no.2
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    • pp.26.1-26.11
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    • 2023
  • Background: Angiotensin-converting enzyme inhibitor (ACEi) inhibits the catalysis of angiotensin I to angiotensin II and the degradation of substance P (SP) and bradykinin (BK). While the possible relationship between ACEi and SP in nociceptive mice was recently suggested, the effect of ACEi on signal transduction in astrocytes remains unclear. Objectives: This study examined whether ACE inhibition with captopril or enalapril modulates the levels of SP and BK in primary cultured astrocytes and whether this change modulates PKC isoforms (PKCα, PKCβI, and PKCε) expression in cultured astrocytes. Methods: Immunocytochemistry and Western blot analysis were performed to examine the changes in the levels of SP and BK and the expression of the PKC isoforms in primary cultured astrocytes, respectively. Results: The treatment of captopril or enalapril increased the immunoreactivity of SP and BK significantly in glial fibrillary acidic protein-positive cultured astrocytes. These increases were suppressed by a pretreatment with an angiotensin-converting enzyme. In addition, treatment with captopril increased the expression of the PKCβI isoform in cultured astrocytes, while there were no changes in the expression of the PKCα and PKCε isoforms after the captopril treatment. The captopril-induced increased expression of the PKCβI isoform was inhibited by a pretreatment with the neurokinin-1 receptor antagonist, L-733,060, the BK B1 receptor antagonist, R 715, or the BK B2 receptor antagonist, HOE 140. Conclusions: These results suggest that ACE inhibition with captopril or enalapril increases the levels of SP and BK in cultured astrocytes and that the activation of SP and BK receptors mediates the captopril-induced increase in the expression of the PKCβI isoform.

Effects of Follicle Cells on the Chymotrypsin Resistance of Mouse Oocytes (난포세포가 생쥐 난자의 Chymotrypsin에 대한 내성에 미치는 영향)

  • Kim, Seong-Im;Bae, In-Ha;Kim, Hae-Kwon;Kim, Sung-Rye
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.407-417
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    • 1999
  • Objective: Mammalian follicle cells are the most important somatic cells which help oocytes grow, mature and ovulate and thus are believed to provide oocytes with various functional and structural components. In the present study we have examined whether cumulus or granulosa cells might playa role in establishing the plasma membrane structure of mouse oocytes during meiotic maturation. Design: In particular the differential resistances of mouse oocytes against chymotrypsin treatment were examined following culture with or without cumulus or granulosa cells, or in these cell-conditioned media. Results: When mouse denuded oocytes, freed from their surrounding cumulus cells, were cultured in vitro for $17{\sim}18hr$ and then treated with 1% chymotrypsin, half of the oocytes underwent degeneration within 37.5 min ($t_{50}=37.5{\pm}7.5min$) after the treatment. In contrast cumulus-enclosed oocytes showed $t_{50}=207.0$. Similarly, when oocytes were co-cultured with cumulus cells which were not associated with the oocytes but present in the same medium, the $t_{50}$ of co-cultured oocytes was $177.5{\pm}13.1min$. Furthermore, when oocytes were cultured in the cumulus cell-conditioned medium, $t_{50}$ of these oocytes was $190.0{\pm}10.8min$ whereas $t_{50}$ of the oocytes cultured in M16 alone was $25.5{\pm}2.9min$. Granulosa cell-conditioned medium also increased the resistance of oocytes against chymotrypsin treatment such that $t_{50}$ of oocytes cultured in granulosa cell-conditioned medium was $152.5{\pm}19.0min$ while that of oocytes cultured in M16 alone was $70.0{\pm}8.2min$. To see what molecular components of follicle cell-conditioned medium are involved in the above effects, the granulosa cell-conditioned medium was separated into two fractions by using Microcon-10 membrane filter having a 10 kDa cut-off range. When denuded oocytes were cultured in medium containing the retentate, $t_{50}$ of the oocytes was $70.0{\pm}10.5min$. In contrast, $t_{50}$ of the denuded oocytes cultured in medium containing the filtrate was $142.0{\pm}26.5min$. $T_{50}$ of denuded oocytes cultured in medium containing both retentate and filtrate was $188.0{\pm}13.6min$. However, $t_{50}$ of denuded oocytes cultured in M16 alone was $70.0{\pm}11.0min$ and that of oocytes cultured in whole granulosa cell-conditioned medium was $156.0{\pm}27.9min$. When surface membrane proteins of oocytes were electrophoretically analyzed, no difference was found between the protein profiles of oocytes cultured in M16 alone and of those cultured in the filtrate. Conclusions: Based upon these results, it is concluded that mouse follicle cells secrete a factor(s) which enhance the resistance of mouse oocytes against a proteolytic enzyme treatment. The factor appears to be a small molecules having a molecular weight less than 10 kDa.

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Food Components of Wild and Cultured Fresh Water Fishes (천연 및 양식산 담수어의 식품성분)

  • KIM Kyung-Sam;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.195-211
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    • 1986
  • The object of this study is to obtain fundamental data on cultured fishes produced in Korea to improve their food components. For this purpose, the food components of cultured fresh water fishes such as eel, Anguilla japonica, snakehead, Channa argus, and common carp, Cyprinus carpio, were investigated and compared with those of the wild ones. The results obtained are summarized as follows: 1. Common characteristics in the proximate composition were that wild fish was higher in crude protein content and lower in crude lipid content than those of cultured one. 2. Among the 9 kinds of minerals analyzed in all the samples, sodium, potassium, calcium and magnesium contents were absolutely predominant being more than $99.52\%$. These four elements in feedstuff also occupied $99.68{\sim}99.92%$ of total minerals. 3. The neutral lipids of wild and cultured eel, snakehead and common carp occupied $55.7{\sim}95.8%$ of lipid fractions, while the content of the phospholipids in snakehead was particularly higher than those of others. 4. The neutral lipids of wild and cultured eel, snakehead and common carp mainly consisted of triglycerides ($85{\sim}95%$), and a little quantity of diglycerides, monoglycerides, free sterol ester and hydrocarbon were also identified in the neutral lipid. 5. The phospolipids of eel and common carp were mainly occupied by phosphatidyl choline ($71.3{\sim}83.9%$), followed by phosphatidyl ethanolamine ($12.1{\sim}23.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). The phospholipids of snakhead consisted of phosphatidyl choline ($50.7{\sim}64.5%$), phosphatidyl ethanolamine ($28.0{\sim}35.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). Generally, phosphatidyl choline content was higher in wild fish than in cultured one, while phosphatidyl ethanolamine and phosphatidyl serine contents were higher in cultured one. 6. The major fatty acids in total lipid of wild eel, snakehead and common carp were $C_{16:0}\;and\;C_{20:5}$, while those in cultured ones were $C_{18:1},\;C_{18:2}\;and\;C_{22:6}$. The fatty acid composition of neutral lipids showed similar tendency to that of total lipid, and the main fatty acids in phospholipids of cultured fishes were $C_{18:1}\;and\;C_{18:2}$. In glycolipids, $C_{20:5}\;and\;C_{22:6}$ were higher in wild fishes, while $C_{18:2}$ were higher in cultured ones. 7. Total amino acids contents of wild and cultured eel were nearly the same, being $16.65\%$ ana $15.99\%$ respectively. The major amino acids of wild and cultured fish were glutamic acid, leucine, aspartic acid and lysine in order. In snakehead, the contents of aspartic acid and proline in cultured fish were higher than those in wild one, while the contents of glutamic acid, alanine, glycine were higher in the wild one. Total amino acid content of cultured common carp was $21.7\%$ compared with $17.08\%$ in wild one. The contents of glutamic acid, aspartic acid, glycine, proline and alanine occupied higher quantities in cultured common carp compared with those in wild one while the other amino acids revealed no significant difference. 8. Aspartic acid in free amino acids of cultured eel held $1.0\%$ of total free amino acids, while that in wild eel held $2.9\%$. Histidine, arginine and tyrosine content of cultured fish were two times higher than those of wild one. But free amino acid composition of samples seemed to be no marked differences according to cultured places. The contents of arginine, aspartic acid, glutamic acid, methionine and phenylalanine of snakehead ware higher in wild one than in cultured one, while the contents of lysine, histidine, glycine, and alanine ware higher in cultured one. In free amino acids content of wild common carp, histidine, glycine and lysine occupied $76.9\%$ of total free amino acids. Lysine, histidine, aspartic acid, alanine, valine and leucine were higher in wild one compared with those of cultured one, while glycine and tyrosine contents were higher in cultured fish.

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