• 제목/요약/키워드: Co-incubation

검색결과 508건 처리시간 0.031초

흰쥐 해마 절편에서 산소고갈에 의한 [$^3H$-5-hydroxytryptamine 유리변동에 미치는 포도당고갈의 영향 (The Effect of Glucose Deprivation on the Oxygen Deprivation-induced Changes of [[$^3H$]-5-hydroxytryptamine Release in Rat Hippocampal Slices)

  • 이경은
    • Toxicological Research
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    • 제14권4호
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    • pp.483-488
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    • 1998
  • During cerebral ischemia two important factors such as hypoxia and reduction of glucose can act as modulating stressor affecting the release of amine neurotransmitters including 5-hydroxytryptamine (5-HT). This study was performed to investigate the effect of glucose deprivation on the oxygen deprivation-induced changes of [3H]-5-HT release in the rat hippocampal slices. Experimental groups were divided into 4 groups for this study: normoxic/normoglycemic group, oxygen-deprived group, glucose-deprived group, and oxygen/glucose-deprived group. The hippocampus of rat brain was sliced by 400 $\mu\textrm{m}$ thickness with manual chopper. After 30 minutes preincubation in the normal buffer, the slices were incubated for 20 min in buffer containing [3H]-5-HT (0.1 M, 74 $\mu\textrm$Ci) for uptake. To measure the release of [3H]-5-HT into the buffer, the incubation medium was drained of and refilled with fresh buffer every ten minutes through a sequence of 14 tubes. Oxygen deprivation by gassing with 95% $N_2$/5% $CO_2$ and/or glucose deprivation was done in the 6th and 7th tube. The radioactivities in each buffer and the tissue were counted using scintillation counter. The results were expressed as fractional release. When slices were exposed to oxygen-deprived media for 20 min, the diminution followed by the rebound release of [3H]-5-HT was observed during the post-oxygen deprived period. However, glucose deprivation or oxygen/glucose deprivation markedly increased the release of [3H]-5-HT. which was opposite to the pattern observed in oxygen-deprived group. These results suggested that oxygen deprivation itself inhibits [3H]-5-HT release in rat hippocampal slices during oxygen-deprived period, but additional glucose deprivation convert the inhibitory response to increase of [3H]-5-HT release.

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식품소재로서의 한천올리고당의 이화학적 특성 (Physicochemical Properties of Agarooligosaccharides for Using as Food Stuffs)

  • 김봉조;송창문;하순득;황선희;김학주;배승권;공재열
    • 한국식품과학회지
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    • 제32권2호
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    • pp.284-290
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    • 2000
  • 한천분해능이 뛰어난 해양세균 Bacillus cereus ASK202를 사용하여 최적조건 하에서 배양실험을 행한 결과, 배양시간 30시간 경과시에 최대량의 한천분해효소가 생산되었으며, 그 생산량은 160.8 unit/L로 기존에 보고된 다른 균주들에 비해 한천분해효소 생산능이 뛰어남을 확인하였다. 또한, 생산된 한천분해효소를 이용하여 기질인 한천과 반응시킨 결과, 주로 중합도 2, 4, 6의 올리고당이 생성됨을 확인하였다. 그리고, 한천올리고당의 물리 화학적 특성을 기존의 제품화된 각종 올리고당과 비교 분석한 결과, 고점성, 높은 올리고당의 함유율, 뛰어난 내산 내열성, 저 감미도를 확인하였다.

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조직 재생을 위한 Poly (D, L-lactide-co-glycolide) 표면에 $(1{\to}3)(1{\to}6)-{\beta}-glucan$ 고정에 대한 세포 점착 및 성장 효과 (Surface Immobilization of $(1{\to}3)(1{\to}6)-{\beta}-glucan$ onto Biodegradable Polymer for Tissue Regeneration)

  • 이상길;이정복;유성미;박종철;최재봉;김정구
    • 대한의용생체공학회:의공학회지
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    • 제27권5호
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    • pp.218-223
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    • 2006
  • We examined the effects of ${\beta}$-glucan-reinforced PLGA film and scaffold on HDFs (human dermal fibroblast) attachment and proliferation. The PLGA films were prepared by simple solvent-casting method. The prepared films were grafted with $(1{\to}3)(1{\to}6)-{\beta}-glucan$ in various ratios after plasma treatment on surface. The surface of the film was characterized by contact angle measurement, scanning electron microscope (SEM), and Fourier-transform infrared spectrophotometer (FT-IR). The amount of $(1{\to}3)(1{\to}6)-{\beta}-glucan$ in the prepared film was indirectly determined by phenol-sulfuric acid method. The HDFs (Human dermal fibroblasts) were used to evaluate the cell attachment and proliferation on PLGA specimens before and after plasma/${\beta}-glucan$ treatment. The result showed that the plasma treated groups exhibited more mont of ${\beta}-glucan$ might be grafted than the non plasma treated groups. Cell attachment was significantly enhanced in the plasma/${\beta}-glucan$ grafted group after 4 hours incubation (p<0.05) due to the improved hydrophilicity and cytoactivity effect of the ${\beta}-glucan$. The cell proliferation of plasma/${\beta}-glucan$ (2mg/ml) grafted group was the highest rate among the groups (p<0.05).

레진 계열 근관봉함재 Adseal의 세포독성에 관한 연구 (CYTOTOXICITY OF RESIN-BASED ROOT CANAL SEALER, ADSEAL)

  • 김희정;백승호;이우철;박한수;배광식
    • Restorative Dentistry and Endodontics
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    • 제29권6호
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    • pp.498-503
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    • 2004
  • The properties of ideal root canal sealers include the ability of sealing the total root canal system and no toxic effects to periradicular tissues. Cytotoxicity test using cell culture is a common screening method for evaluation of the biocompatibility of root canal sealers. The purpose of this study was to investigate the cytotoxic effect of newly developed resin-based sealer (Adseal 1, 2, and 3) comparing with those commercial resin-based sealers (AH26 and AH Plus), ZOE-based sealers (Tubliseal EWT, Pulp Canal Sealer EWT) and calcium hydroxide based sealer (Sealapex), An indirect contact test of cytotoxicity by agar diffusion was performed according to the international standard ISO 10993-5. L929 fibroblast cells were incubated at $37^{\circ}C$ in humidified 5% $CO_2-containing$ air atmosphere. The freshly mixed test materials were inserted into glass rings of internal diameter 5 mm and height 5 mm placed on the agar. After the 24 hrs incubation period, the decolorization zones around the test materials were assessed using an inverted microscope with a calibrated screen. A Decolorization Index was determined for each specimen. Adseal 1. 2, and 3 did not exert any cytotoxic effects, whereas AH26, AH Plus, Tubliseal EWT, Pulp Canal Sealer EWT, and Sealapex produced mild cytotoxicity.

Korean Red Ginseng protects endothelial cells from serum-deprived apoptosis by regulating Bcl-2 family protein dynamics and caspase S-nitrosylation

  • Kim, Young-Mi;Kim, Jung Hwan;Kwon, Hyuk Min;Lee, Dong Heon;Won, Moo-Ho;Kwon, Young-Guen;Kim, Young-Myeong
    • Journal of Ginseng Research
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    • 제37권4호
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    • pp.413-424
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    • 2013
  • Korean Red Ginseng extract (KRGE) is a traditional herbal medicine utilized to prevent endothelium dysfunction in the cardiovascular system; however, its underlying mechanism has not been clearly elucidated. We here examined the pharmacological effect and molecular mechanism of KRGE on apoptosis of human umbilical vein endothelial cells (HUVECs) in a serum-deprived apoptosis model. KRGE protected HUVECs from serum-deprived apoptosis by inhibiting mitochondrial cytochrome c release and caspase-9/-3 activation. This protective effect was significantly higher than that of American ginseng extract. KRGE treatment increased antiapoptotic Bcl-2 and Bcl-$X_L$ protein expression and Akt-dependent Bad phosphorylation. Moreover, KRGE prevented serum deprivation-induced subcellular redistribution of these proteins between the mitochondrion and the cytosol, resulting in suppression of mitochondrial cytochrome c release. In addition, KRGE increased nitric oxide (NO) production via Akt-dependent activation of endothelial NO synthase (eNOS), as well as inhibited caspase-9/-3 activities. These increases were reversed by co-treatment of cells with inhibitors of eNOS and phosphoinositide 3-kinase (PI3K) and pre-incubation of cell lysates in dithiothreitol, indicating KRGE induces NO-mediated caspase modification. Indeed, KRGE inhibited caspase-3 activity via S-nitrosylation. These findings suggest that KRGE prevents serum deprivation-induced HUVEC apoptosis via increased Bcl-2 and Bcl-$X_L$ protein expression, PI3K/Akt-dependent Bad phosphorylation, and eNOS/NO-mediated S-nitrosylation of caspases. The cytoprotective property of KRGE may be valuable for developing new pharmaceutical means that limit endothelial cell death induced during the pathogenesis of vascular diseases.

Intramolecular Disulfide Bonds for Biogenesis of Calcium Homeostasis Modulator 1 Ion Channel Are Dispensable for Voltage-Dependent Activation

  • Kwon, Jae Won;Jeon, Young Keul;Kim, Jinsung;Kim, Sang Jeong;Kim, Sung Joon
    • Molecules and Cells
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    • 제44권10호
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    • pp.758-769
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    • 2021
  • Calcium homeostasis modulator 1 (CALHM1) is a membrane protein with four transmembrane helices that form an octameric ion channel with voltage-dependent activation. There are four conserved cysteine (Cys) residues in the extracellular domain that form two intramolecular disulfide bonds. We investigated the roles of C42-C127 and C44-C161 in human CALHM1 channel biogenesis and the ionic current (ICALHM1). Replacing Cys with Ser or Ala abolished the membrane trafficking as well as ICALHM1. Immunoblotting analysis revealed dithiothreitol-sensitive multimeric CALHM1, which was markedly reduced in C44S and C161S, but preserved in C42S and C127S. The mixed expression of C42S and wild-type did not show a dominant-negative effect. While the heteromeric assembly of CALHM1 and CALHM3 formed active ion channels, the co-expression of C42S and CALHM3 did not produce functional channels. Despite the critical structural role of the extracellular cysteine residues, a treatment with the membrane-impermeable reducing agent tris(2-carboxyethyl) phosphine (TCEP, 2 mM) did not affect ICALHM1 for up to 30 min. Interestingly, incubation with TCEP (2 mM) for 2-6 h reduced both ICALHM1 and the surface expression of CALHM1 in a time-dependent manner. We propose that the intramolecular disulfide bonds are essential for folding, oligomerization, trafficking and maintenance of CALHM1 in the plasma membrane, but dispensable for the voltage-dependent activation once expressed on the plasma membrane.

Effects of maternal dietary energy restriction on breast muscle fibre development in the offspring of broiler breeders

  • Wu, Hongzhi;Sun, Hao;Ma, Chengzhan;Lian, Lina;Lu, Lei;Xu, Liangmei;Xu, Li
    • Animal Bioscience
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    • 제34권11호
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    • pp.1829-1838
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    • 2021
  • Objective: The effects of maternal dietary energy levels on breast muscle fibre development in offspring of broiler breeders were investigated. Methods: A total of 480 20-week-old Arbor Acres (AA) healthy female broiler breeders, with an average body weight of 2.33±0.01 kg, were randomly divided into 4 treatment groups with 6 replicates and 20 broiler breeders for each replicate and fed a corn and soybean meal diet with 100%, 80%, 70%, and 50% energy levels, respectively. Approximately 300 eggs per treatment were collected for incubation for 6 days. Then, 120 0-day-old female broilers at each energy level were randomly selected and divided into 6 replicates with 20 broilers for each replicate, with this experimental phase with the offspring lasting for 49 days. Results: Compared with the 100% energy group, the breast muscle fibre diameter at embryonic day 21 in the 80% energy group was significantly reduced (p<0.05). In the 80% energy group, the muscle fibre density of the breast increased significantly (p<0.05) at embryonic days 15 and 21. The breast muscle fibre diameter of the offspring in each group was significantly decreased (p<0.05) on the 1st day. The breast muscle sarcomere length of the embryos in the 80% energy group was significantly higher (p<0.05) than those in the 70% and 50% energy groups. Compared with the 100% energy group, the expression of the myostatin gene in the offspring was significantly decreased (p<0.05). Conclusion: In conclusion, the effects of a maternal dietary energy level of 80% in this study were found to be optimal for breast muscle fibre development in offspring, which indicated that the metabolic energy level of AA broilers of 9.36 MJ/kg for the mid-term diet for laying eggs has a more practical significance.

보골탕이 Monosodium Iodoacetate 유도 골관절염과 Interleukin-1β 유도 연골세포에 미치는 보호 효과 (Protective Effects of Bogol-tang on Monosodium Iodoacetate-induced Osteoarthritis and Interleukin-1β-treated Primary Chondrocytes)

  • 성진욱;이해웅;강경화;김경민;조성우
    • 한방재활의학과학회지
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    • 제29권2호
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    • pp.101-113
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    • 2019
  • Objectives Bogol-tang has clinically been used to protect joint cartilage and to treat osteoarthritis. Our objective was to study the protective effect of Bogol-tang extract (BGT) in functional impairment, behavioral disorders, cartilage loss and pathological changes in a monoiodoacetate (MIA)-induced murine osteoarthritis (OA) model and interleukin (IL)-$1{\beta}$ -treated primary rat chondrocytes. Methods Mouse knee joints were injected with MIA, a chemical that inhibits glycolysis and causes joint inflammation and matrix loss. MIA-OA induced mice orally administered BGT or acetaminophen (AAP) for 18 days by daily. Primary rat chondrocytes were pretreated with BGT or dexamethasone (DEX) and followed by co-incubation with IL-$1{\beta}$ (10 ng/mL). Results In MIA-OA mice model, BGT led to delayed response on hot plate analysis, and suppressed the cartilage loss and damages in joint tissues. BGT suppressed the elevated levels of inflammatory mediators, nitrite and $PGE_2$, the gene expression of matrix degrading enzymes, and extracellular-signal-regulated kinases 1/2 and c-JunN-terminal kinase phosphorylation in IL-$1{\beta}$-treated primary rat chondrocytes. Conclusions Our results suggest that BGT improve the knee joint function and delay the cartilage damages by anti-nociceptive, anti-inflammatory and ant-catabolic effects, which indicate BGT could be a potential candidate for osteoarthritis treatment.

A study on comparative feeding value of corn flakes according to temperature and retention time in the pressurized steam chamber

  • Ahn, Jun Sang;Shin, Jung Suh;Kim, Min Ji;Son, Gi Hwal;Kwon, Eung Gi;Shim, Jae Yoon;Kim, Il Young;Cho, Sung Myoun;Cho, Sang Rae;Park, Byung Ki
    • Journal of Animal Science and Technology
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    • 제61권3호
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    • pp.170-181
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    • 2019
  • This study aimed to investigate the effects of temperature and retention time of the pressurized steam chamber on the ruminal fermentation characteristics and nutrient degradability of corn flakes in three Korean native Hanwoo cows and three Holstein cows implanted with a ruminal fistula. Corn kernels were categorized into 13 groups based on the chamber temperature (range, $100^{\circ}C-116^{\circ}C$) and retention time (range, 700-950 s). The pH value was lowest in T1 regardless of breed. Propionate concentration was the highest in T2 (p < 0.05). Total-volatile fatty acid (VFA) concentration was slightly but not significantly greater in T2 than in other conditions. Dry matter (p < 0.05), starch, and crude protein (p < 0.05) degradability were the highest in T1. At different incubation times and with different breeds, dry matter, starch, and crude protein degradability of corn flakes were the highest in T1. Thus, the present results indicate that the optimal temperature and retention time of the pressurized steam chamber should be $100^{\circ}C-105^{\circ}C$ and 700-720 s.

능성어 (Hyporthodus septemfasciatus) ♀×대왕바리 (Epinephelus lanceolatus) ♂ 수정란의 난발생 (Embryonic Development of Fertilized Eggs of Convict Grouper (Hyporthodus septemfasciatus) ♀×Giant Grouper (Epinephelus lanceolatus) ♂)

  • 노충환;윤낙진
    • 한국어류학회지
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    • 제31권1호
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    • pp.23-29
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    • 2019
  • 이번 연구에서 바리과 어종의 새로운 교잡종을 생산하기 위한 연구의 일환으로 능성어 (Hyporthodus septemfasciatus) ♀${\times}$대왕바리 (Epinephelus lanceolatus) ♂ (CGGG)의 발생 양상, 부화율, 기형 발생률 그리고 부화 소요시간을 조사하고, 교잡종의 모계어종의 순종 (능성어 ♀${\times}$♂, CG)과 비교하였다. CGGG와 CG 수정란은 자외선 살균 해수가 채워진 5-L 용량의 비이커 9개에 3,000~5,000개/비이커의 밀도로 수용되었으며, 발생 기간 중 수온은 $23.5{\sim}24.8^{\circ}C$ 그리고 염분은 32.1~32.8 ppt 였다. CGGG의 수정률과 부화율은 각각 $69.4{\pm}1.5%$$59.0{\pm}5.1%$로서 CG에 비해 유의하게 낮았다 (p<0.05). CGGG는 비균등 난할, 배체 체형 기형 그리고 부화자어의 척추 기형이 높게 출현한 것을 제외하고, CG와 동일한 난발생 양상을 보였다. CGGG의 부화까지 소요 시간은 CG에 비해 약 2시간 느렸다(p<0.05). 이상의 결과에서 CGGG는 수정률, 부화율 그리고 기형률 등 발생능력이 CG에 비해 낮았지만 부화자어의 대량생산이 가능할 것으로 판단된다.