• Title/Summary/Keyword: Co-incubation

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Studies on the Appearance and Function of the Central Cavity of Corpus Luteum in the Ovaries of the Korean Native Cattle (한우 난소의 중심강이 있는 황체의 출현과 기능에 관한 연구)

  • 성환후;오성종;양보석;백광수;곽수동;정진관
    • Korean Journal of Animal Reproduction
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    • v.18 no.4
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    • pp.245-250
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    • 1995
  • The appearance and function of corpora lutea(CL) with a central cavity in the ovaries of Korean Native Cattle (KNC) were investigated endocrinologically and histochemically. The CL were enucleated from KNC ovaries within 2~3 hrs local slaughter house and classified with central cavity CL or not. Enzymatically dispersed luteal cell (1$\times$106 live cell/ml of Dulbecco's Modified Eagle Media) with or without cavity of CL cultured at 37$^{\circ}C$ in a humidified incubation (5% CO2 : 95% air) for 72hr. A central cavity in the CL of KNC was found in 58.8% of CL-1, 34.9% of CL 2, 39.1% of CL-3, and 11.1% of CL-4, respectively. There were no significant difference between protein content of CL with and without a central cavity. Mean progesterone secretion after 36h of in vitro luteal cell culture were significantly (p<0.05) higher in CL with central cavity than without cavity. However, the luteal cavitic wall was composed of the connective tissue band of the reticular and collagen fibers and then these connective tissue band extended into the CL with cavity. These results suggest that the central cavity of CL may be caused infertility in KNC.

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Isolation and Identification of Mycobactin Dependent Acid-fast Bacteria (M. paratuberculosis) from Bovine Fecal Material (우유래(牛由來) Mycobactin의존성(依存性) 항산성세균(抗酸性細菌) (M. paratuberculosis)의 분리동정(分離同定))

  • Jeon, Yun-seong;Lee, Bang-whan;Kim, Jong-bae;Choi, Chul-soon;Kim, Jin-koo
    • Korean Journal of Veterinary Research
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    • v.24 no.1
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    • pp.58-63
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    • 1984
  • Fecal material from cattle, which was confirmed to be infected with Johne's disease by clinical and pathological symptoms, was decontaminated with 4% NaOH and inoculated into the $L{\ddot{o}}wenstein$-Jensen media supplemented with 1% of heat-killed Mycobacterium bovis. After 2-4 week-incubation at $37^{\circ}C$, typical acid-fast mycobacteria was isolated. With the results of staining properties, morphological characteristics, the requirement of mycobactin for growth and the other biochemical properties, isolated mycobacteria was identified as Mycobacterium paratuberculosis. Female guinea pigs were sensitized with the isolates, and skin test was done with purified protein derivatives (PPDs) of M. avium, M. bovis and M. paratuberculosis 4 weeks after sensitization. Animals showed the largest reaction to the PPDs of M. avium and M. paratuverculosis.

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An Intersex with Os Clitoris and Ovotestes in Korean Native Jindo Dog (진도개에서 발생한 Os clitoris와 Ovotestes를 특징으로 하는 양성자 사례 연구)

  • Cho, Sung-Jin;Lee, Hyun-A;Park, Sang-Ho;Kim, Ok-Jin
    • Journal of Veterinary Clinics
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    • v.27 no.1
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    • pp.83-87
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    • 2010
  • Hermaphroditism occurs rarely in human and animal populations. Previously, in Korea, The hermaphrodite dog in American Cocker Spaniel dogs have been reported in six cases. However, in this case, we found a hermaphrodite dog in Korean native Jindo dog. The characteristics were its enlarged os clitoris that was shown a reddish fingerlike structure protruding from the vulva. The dog was diagnosed as hermaphroditism with ovotestes. To our knowledge, this report is the first case of hermaphroditism in Korean native dog.

The Possible Participation of the Mesophyll on Stomatal Opening

  • Lee, Joon-Sang
    • Korean Journal of Environmental Biology
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    • v.19 no.1
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    • pp.1-6
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    • 2001
  • Many researchers have been studied with guard cell protoplasts and detached epidermis as they think that properly stabilized protoplasts and detached epidermis retain many of the properties of intact guard cells. However, some studies have shown that stomata in detached epidermis behave differently, both quantitatively and qualitatively, from those in the intact leaf. Stomata in the intact leaf are very sensitive to environmental factors such as light, $CO_2$ and osmotic stress, but stomata in detached epidermis are less sensitive to these factors than those in the intact leaf. The clearest evidence to suggest the different response between detached epidermis and intact leaf obtained from the experiments with heavy metal, cadmium. 3-weeks old Commelina. communis was transferred to and grown in Hoagland solution in the presence or absence of 5 mM $Cd^{2+}$ for 4 days. The application of $Cd^{2+}$ showed about 70% inhibition of stomatal conductance when measured at various light intensity (100-1,000 $\mu$mole $m^{-2}s^{-1}). However, stomata in detached epidermis floated on an incubation medium containing 100 $\mu$M $Cd^{2+}$ opened to a degree of about 8.38 fm, but the stomata treated with no cadmium opened to 3.74 ${\mu}{\textrm}{m}$. These results were unexpected as the intact leaf grown in a Hoagland solution containing cadmium showed very negative physiological responses. These results showed that stomata in detached epidermis and in the intact leaf could respond reversely. Therefore, it is possible that we now misunderstand how stomata open in real natural condition.

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Nanofabrication of Microbial Polyester by Electrospinning Promotes Cell Attachment

  • Lee, Ik-Sang;Kwon, Oh-Hyeong;Wan Meng;Kang, Inn-Kyu;Yoshihiro Ito
    • Macromolecular Research
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    • v.12 no.4
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    • pp.374-378
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    • 2004
  • The biodegradable and biocompatible poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a copolymer of microbial polyester, was fabricated as nanofibrous mats by electrospinning. Image analysis of the electrospun nanofibers fabricated from a 2 wt% 2,2,2-trifluoroethanol solution revealed a unimodal distribution pattern of fiber diameters with an observed average diameter of ca. 185 nm. The fiber diameter of electrospun fabrics could be controlled by adjusting the electro spinning parameters, including the solvent composition, concentration, applied voltage, and tip-to-collector distance. Chondrocytes derived from rabbit ear were cultured on a PHBV cast film and an electrospun PHBV nano-fibrous mat. After incubation for 2 h, the percentages of attached chondrocytes on the surfaces of the flat PHBV film and the PHBV nanofibrous mat were 19.0 and 30.1 %, respectively. On the surface of the electrospun PHBV fabric, more chondrocytes were attached and appeared to have a much greater spreaded morphology than did that of the flat PHBV cast film in the early culture stage. The electro spun PHBV nanofabric provides an attractive structure for the attachment and growth of chondrocytes as cell culture surfaces for tissue engineering.

Variations of Abundance and Hatch Timing of Dungeness Crab Larvae in Southeastern Alaska: Implications for Climate Effect

  • Park, Won-Gyu;Shirley, Thomas C.
    • Animal cells and systems
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    • v.12 no.4
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    • pp.287-295
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    • 2008
  • Variations of larval abundance and hatch timing of Dungeness crabs, Cancer magister Dana 1852, were investigated. Dungeness crab larvae were monthly collected at 16 stations arrayed in four transects, Upper Chatham, Icy Strait, Cross Sound, and Icy Point, in southeastern Alaska from May to September 1997-2004. Larval abundance at all transects was the highest in June except in the Icy Point transect. Larval abundance was the highest in the Icy Strait transect, moderate in the Upper Chatham and Cross Sound transects, and the lowest in the Icy Point transect. Zoeae I(ZI) was predominated in May; thereafter ZI decreased and late zoeal stages occurred. In May and June, small numbers of late stage larvae unusually co-occurred with ZI in three transects. These late stage larvae may have been transported from where hatching occurs earlier. The timing of ZI occurrence varied interannually and was related to degreedays during the egg incubation period of Dungeness crabs: later larval hatching in 1997 and 2002 when temperatures were colder, while earlier larval hatching in 1998 when temperatures were warmer. The distribution patterns of Dungeness crab larvae in southeastern Alaska were markedly different from those reported from other areas of the species distribution ranges: larvae occurring much later in the year, and late stage larvae occurring in inland waters.

The Degradation of Chitin with Food Grade Papain

  • Han, Beom-Ku;You, Tak;Moon, Jong-Kook;Kim, Sae-Bom;Jo, Do-Hyun
    • Journal of Applied Biological Chemistry
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    • v.43 no.4
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    • pp.246-249
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    • 2000
  • We investigated the production of chitin oligosaccharides using food grade papain. A solution of commercial food grade papain (FGP) was dialyzed for 12 h before measuring its chitinolytic activity. The effects of enzyme concentration, reaction temperature, and pH on the endochitinase and $\beta$-N-acetylglucosaminidase activities and the thermostability of these enzymes were investigated. In adddition, the reaction products were analyzed with gel filtration on a Bio-Gel P2. The endochitinase activity was twentyfold higher than that of $\beta$-N-acetylglucosaminidase. The optimal endochitinase activity was at pH 3.0, while the maximal $\beta$-N-acetylglucosaminidase activity was at pH 6.0. The reaction product consisted mainly of the dimer of N -acetylglucosamine, with a small amount of its trimer. Under the experimental conditions, $120{\mu}g$ of chitin oligomers were obtained with 1 mg of FGP protein after an incubation of 2 h.

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Production of Liquiritigenin with Cell-based Biotransformation and Its Anti-Aging Activity (균사체 생물전환기술을 이용한 리퀘리티게닌 생산과 항노화 활성)

  • Hwang, Hye Jin;Jeong, Sang Chul;Park, Jong Pil
    • KSBB Journal
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    • v.30 no.4
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    • pp.166-174
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    • 2015
  • In this study, an efficient whole cell-based biotransformation for the production of liquiritigenin was developed using Laetiporus sulphureus CS0218 as biocatalyst and aqueous extracts of Glycyrrhiza uralensis as co-substrate, respectively. In order to determine the efficacy of this method, the optimal bioconversion conditions including mycelial growth, three important enzyme activities (${\beta}$-glucosidase, ${\alpha}$-rhamnosidase and ${\beta}$-xylosidase), and apparent viscosity of culture broth were monitored. After optimization, aqueous extracts of G. uralensis were added to the culture medium to directly produce algycone liquiritigenin. By applying this strategy, 67.5% of liquiritin was converted to liquiritigenin at pH 3.0 after 9 days of incubation and finally liquiritigenin was purified from the reaction mixture. And then, their biological activities including anti-oxidant and superoxide dismutase were observed. In fact, purified liquiritigenin was capable of bi-directional functions (i.e., either up-regulation or down-regulation of SIRT1 which is associated with aging). The results indicate that this strategy would be beneficial to produce biologically active liquiritigenin and could be used in pharmaceutical, cosmetic and food applications.

Enhanced sialylation and in vivo efficacy of recombinant human α-galactosidase through in vitro glycosylation

  • Sohn, Youngsoo;Lee, Jung Mi;Park, Heung-Rok;Jung, Sung-Chul;Park, Tai Hyun;Oh, Doo-Byoung
    • BMB Reports
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    • v.46 no.3
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    • pp.157-162
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    • 2013
  • Human ${\alpha}$-galactosidase A (GLA) has been used in enzyme replacement therapy for patients with Fabry disease. We expressed recombinant GLA from Chinese hamster ovary cells with very high productivity. When compared to an approved GLA (agalsidase beta), its size and charge were found to be smaller and more neutral. These differences resulted from the lack of terminal sialic acids playing essential roles in the serum half-life and proper tissue targeting. Because a simple sialylation reaction was not enough to increase the sialic acid content, a combined reaction using galactosyltransferase, sialyltransferase, and their sugar substrates at the same time was developed and optimized to reduce the incubation time. The product generated by this reaction had nearly the same size, isoelectric points, and sialic acid content as agalsidase beta. Furthermore, it had better in vivo efficacy to degrade the accumulated globotriaosylceramide in target organs of Fabry mice compared to an unmodified version.

Inhibition of Lipopolysaccaride-induced Inducible Nitric Oxide (iNOS) mRNA Expression and Nitric Oxide Production by Higenamine in Murine Peritoneal Macrophages

  • Lee, Hoi-Young;Lee, Jang-Soon;Kim, Eun-Ju;Han, Jeung-Whan;Lee, Hyang-Woo;Kang, Young-Jin;Chang, Ki-Churl
    • Archives of Pharmacal Research
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    • v.22 no.1
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    • pp.55-59
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    • 1999
  • Nitric oxide synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation in rheumatic and autoimmune diseases. The effects of higenamine, a tetrahydroisoquinoline compound, on induction of NOS by bacterial lipopolysaccaride (LPS) were examined in murine peritoneal macrophages. LPS-induced nitrite/nitrate production was markedly inhibited by higenamine which at 0.01 mM, decreased nitrite/nitrate levels by $48.7{\pm}4.4%$This was comparable to the inhibition of LPS-induced nitrite/nitrate production by tetrandrin ($49.51{\pm}2.02%$). at the same concentration. Northern and Western blot analysis of iNOS expression demonstrated that iNOS expression was significantly attenuated following co-incubation of peritoneal macrophages with LPS (10 $\mu\textrm{g}$/m;; 18hrs) and higenamine (0.001, 0.,01 mM; 18hrs). These results suggest that higenamine can inhibit LPS-induced expression of iNOS mRNA in murine peritoneal macrophages. The clinical implications of these findings remain to be established.

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