• Title/Summary/Keyword: Cell complex

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Ganglioside GD1a Activates the Phosphorylation of EGFR in Porcine Oocytes Maturation in vitro

  • Park, Hyo-Jin;Kim, Jin-Woo;Park, Jae-Young;Yang, Seul-Gi;Jung, Jae-Min;Kim, Min-Ji;Koo, Deog-Bon
    • Journal of Embryo Transfer
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    • v.32 no.1
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    • pp.17-24
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    • 2017
  • Ganglioside GD1a is specifically formed by the addition of sialic acid to ganglioside GM1a by ST3 ${\beta}$-galactoside ${\alpha}$-2,3-sialyltransferase 2 (ST3GAL2). Above all, GD1a are known to be related with the functional regulation of several growth factor receptors, including activation and dimerization of epidermal growth factor receptor (EGFR) in tumor cells. The activity of EGF and EGFR is known to be a very important factor for meiotic and cytoplasmic maturation during in vitro maturation (IVM) of mammalian oocytes. However, the role of gangliosides GD1a for EGFR-related signaling pathways in porcine oocyte is not yet clearly understood. Here, we investigated that the effect of ST3GAL2 as synthesizing enzyme GD1a for EGFR activation and phosphorylation during meiotic maturation. To investigate the expression of ST3GAL2 according to the EGF treatment (0, 10 and 50 ng/ml), we observed the patterns of ST3GAL2 genes expression by immunofluorescence staining in denuded oocyte (DO) and cumulus cell-oocyte-complex (COC) during IVM process (22 and 44 h), respectively. Expression levels of ST3GAL2 significantly decreased (p<0.01) in an EGF concentration (10 and 50 ng/ml) dependent manner. And fluorescence expression of ST3GAL2 increased (p<0.01) in the matured COCs for 44 h. Under high EGF concentration (50 ng/ml), ST3GAL2 protein levels was decreased (p<0.01), and their shown opposite expression pattern of phosphorylation-EGFR in COCs of 44 h. Phosphorylation of EGFR significantly increased (p<0.01) in matured COCs treated with GD1a for 44 h. In addition, ST3GAL2 protein levels significantly decreased (p<0.01) in GD1a ($10{\mu}M$) treated COCs without reference to EGF pre-treatment. These results suggest that treatment of exogenous ganglioside GD1a may play an important role such as EGF in EGFR-related activation and phosphorylation in porcine oocyte maturation of in vitro.

Impacts on Water Quality to an Artificial Lake Due to Sudden Disturbance of Sediments (급격한 저니토 교란이 인공 하구호 수질에 미치는 영향)

  • 서승원;김정훈
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.15 no.1
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    • pp.39-50
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    • 2003
  • This study aims to figure out water quality impacts due to sudden disturbances of sediments during dike construction in land reclamation for the northern part of the Siwhaho Lake where heavily deteriorated settlements from upstream loadings are outstanding. We exploit a 3-D water quality model CE-QUAL-ICM combined with a hydrodynamic model TIDE3D. Simulations are done accounting water-sediment interaction in a 4-$\sigma$ layers. Long-term simulation for 1-year shows that bottom layers around the disturbance location are only affected and marks very high concentration. Complete vertical mixing appears at least 5km apart to downward due to complex effects of geometry, bathymetry and river inflows. It should be addressed that existing condition of the Siwhaho Lake stands for high concentration of COD and TP in winter and spring due to relatively high incoming loadings, however the effect of sediment disturbances yields reverse phenomena, i.e., impacts of dike construction arise greatly in summer and fall. Refined grid system consisting of 150m${\times}$150m rectangular grid, which is doubled system compared to previous study (Suh et al.,2002), gives affordable results by reducing flux differences through a cell especially in front of gate.

A 4 week Randomized, Double-blind Human Trial to Compare the Efficacy and Safety of Aureobasidium pullulans Cultured Solution and Placebo on Improvement of Immune in Subjects (흑효모배양액 분말의 면역관련 사이토카인에 미치는 영향에 대한 무작위 배정 임상연구)

  • Choi, Hae-Yun;Kim, Jong-Dae;Park, Mee-Yeon
    • Korean Journal of Oriental Medicine
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    • v.15 no.3
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    • pp.83-91
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    • 2009
  • $\beta$-glucan is a fiber-type complex sugar (polysaccharide) derived from the cell wall of baker's yeast, oat and barley fiber, and many medicinal mushrooms, such as maitake. The primary uses of $\beta$-glucan are to enhance the immune system, to lower blood cholesterol levels and to treat tumor. $\beta$-glucan has no systemic toxicity in mice, therefore it needed clinical trail to prove efficacy and safety for human. The subjects total 56 healty volunteers were divided into two groups including taken $\beta$-glucan tablet group and placebo group. Subjects were taken two tablets per oral for 4 weeks. They had agreed to take part in this experiment, and didn't take any other clinical trail products. After 4 weeks blood of subjects were checked. The check list are TNF-$\alpha$, INF-$\gamma$, IL-2, IL-4, total WBC, differential WBC, RBC, hemoglobin, platelet, MCV, MCH, MCHC, HCT, Na, K, Ca, Cl, AST, ALT, ALP, $\gamma$-GTP, total protein, triglyceride, total cholesterol, total bilirubin, albumin, uric acid, creatinine, BUN, pH, protein, glucose, ketone body, blood, bilirubin. We evaluated efficacy by cytokines that compare before and after taking. Collected data were analyzed as two sample t-test, chi-square test and ANOVA using SAS V.9.1.This study results are that in TNF-$\alpha$ of $1^{st}$ efficacy measurement item, all of two groups figure were increased significantly compare to before figure. In IL4 of $2^{nd}$ efficacy measurement item, experimental group figure were decreased significantly but placebo group figure were increased. The conclusions show that based on the above results, $\beta$-glucan has favorable effect to enhance immune system, especially IL4 results showed that it has effect to improve the allergic immune system.

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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

The Effects of ATP - sensitive $K^+$ Channel on the Muscle Fatigue in Mouse Skeletal Muscle Cell (골격근 세포에서 ATP-의존성 $K^+$통로의 활성화가 근피로에 미치는 영향)

  • Koo Hyun-Mo;Nam Ki-Won;Kim Suck-Bum;Lee Sun-Min;Kim Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.14 no.2
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    • pp.1-15
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    • 2002
  • Excitation-contraction coupling in skeletal muscle is process by which depolarization of the muscle fiber membrane, elicited by a nerve action potential, triggers the release of $Ca^{2+}$ from the sarcoplasmic reticulum(SR). The resulting rise in intracellular $Ca^{2+}$ concentration$([Ca^{2+}]_i)$ activates the troponin complex, thereby initiating the contraction of the muscle. The question remains as to what factors are involved in the inhibition of SR $Ca^{2+}$ release in fatigued muscle. The purpose of this study was determine whether ATP-sensitive $K^+(K_{ATP})$ channels are activated and contribute to decrease in $[Ca^{2+}]_i$ during fatigue development in the mouse skeletal muscle. To elucidate a role of $K_{ATP})$ in relation to ECC, I measured the modulation effects of $K_{ATP})$ channel blocker(glibenclamide) and opener(pinacidil) on $[Ca^{2+}]_i$ after fatiguing electrical field stimulation(FEFS). Intracellular $Ca^{2+}$ signals were recorded by conforcal laser microscopy(LSM 410) and monitored using the fluorescent $Ca^{2+}$-Sensitive indicator Fluo-3 AM. The results of this study were as followed: 1. The relative [Ca2'li after FEFS in the pre-glibenclamide-treated group was higher than the control. And relative $[Ca^{2+}]_i$ after FEFS in the pre-glibenclamide-treated group was lower than the control. 2. The relative $[Ca^{2+}]_i$ after FEFS for 3 min in the control, pre-glibenclamide-treated group and pre-pinacidil-treated group showed a similar pattern; the gradually significant decrease in $[Ca^{2+}]_i$. But, these decreasing pattern was most significant in the control. These findings suggest a tight relationship between $K_{ATP})$ and $Ca^{2+}$ in ECC during fatigue. Therefore, 1 thought that activation of $K_{ATP})$ channels may be one of mechanisms of the fatigue in skeletal muscle.

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Engraftment of Intraperitoneally Injected Bone Marrow Cells to Newborn Mice Injected with an Angiogenesis Inhibitor (혈관생성 억제제를 주사한 마우스 모델에서의 골수 세포의 복강 내 주입 후 생착)

  • Cho, Su-Jin;Ju, Sun-Young;Woo, So-Youn;Kang, Hyoung-Jin;Ahn, Hyo-Seop;Ryu, Kyung-Ha;Park, Eun-Ae
    • Neonatal Medicine
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    • v.15 no.1
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    • pp.22-31
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    • 2008
  • Purpose : Bronchopulmonary dysplasia (BPD) is characterized by arrested vascular and alveolar growth in the premature lung. Considering the consequences of arrested lung growth, the idea of administering bone marrow cells to enhance the inborn repair mechanism is promising as this may reduce the morbidity and mortality of BPD. We followed enhanced green fluorescent protein (EGFP)-labeled bone marrow cells (BMC) injected intraperitoneally into non-EGFP mice in order to determine their fate after transplantation. Methods : An angiogenesis inhibitor, SU1498, was injected subcutaneously on day 3 in non-EGFP C57BL/6 newborn mice to create a model of arrested alveolar development. On the following day, $1{\times}10^6$ BMCs isolated from major histocompatibility complex (MHC)- matched syngenic EGFP mice were injected intraperitoneally to non-EGFP BPD mice. Morphometric analysis, immunostaining, and confocal microscopy were performed to determine the fate of EGFP-positive stem cells in the injured lung. Results : SU1498 injection reduced alveolar surface area and mean alveolar volume in newborn mice. BMC injection resulted in recovery of lung structure comparable to controls. EGFP-positive BMCs were identified in the lungs of the recipient mice after intraperitoneal injection. The injected EGFP cells were co-stained with endothelial and epithelial cells of the developing lung as determined by confocal microscopy. Conclusion : Our results illustrated that EGFP-positive BMCs engrafted and trans-differentiated into epithelial and endothelial cells after intraperitoneal injection in a mouse model of arrested alveolar development.

Identification of genes involved in inbreeding depression of reproduction in Langshan chickens

  • Xue, Qian;Li, Guohui;Cao, Yuxia;Yin, Jianmei;Zhu, Yunfen;Zhang, Huiyong;Zhou, Chenghao;Shen, Haiyu;Dou, Xinhong;Su, Yijun;Wang, Kehua;Zou, Jianmin;Han, Wei
    • Animal Bioscience
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    • v.34 no.6
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    • pp.975-984
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    • 2021
  • Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.

Next-generation Probiotics, Parabiotics, and Postbiotics (Next-generation probiotics, parabiotics 및 postbiotics)

  • Cho, Kwang Keun;Lee, Seung Ho;Choi, In Soon;Lee, Sang Won
    • Journal of Life Science
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    • v.31 no.6
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    • pp.595-602
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    • 2021
  • Human intestinal microbiota play an important role in the regulation of the host's metabolism. There is a close pathological and physiological interaction between dysbiosis of the intestinal microflora and obesity and metabolic syndrome. Akkermansia muciniphila, which was recently isolated from human feces, accounts for about 1-4% of the intestinal microbiota population. The use of A. muciniphila- derived external membrane protein Amuc_1100 and extracellular vesicles (EVs) could be a new strategy for the treatment of obesity. A. muciniphila is considered a next-generation probiotic (NGP) for the treatment of metabolic disorders, such as obesity. Faecalibacterium prausnitzii accounts for about 5% of the intestinal microbiota population in healthy adults and is an indicator of gut health. F. prausnitzii is a butyrate-producing bacterium, with anti-inflammatory effects, and is considered an NGP for the treatment of immune diseases and diabetes. Postbiotics are complex mixtures of metabolites contained in the cell supernatant secreted by probiotics. Parabiotics are microbial cells in which probiotics are inactivated. Paraprobiotics and postbiotics have many advantages over probiotics, such as clear chemical structures, safe dose parameters, and a long shelf life. Thus, they have the potential to replace probiotics. The most natural strategy to restore the imbalance of the intestinal ecosystem normally is to use NGPs among commensal bacteria in the gut. Therefore, it is necessary to develop new foods or drugs such as parabiotics and postbiotics using NGPs.

Antioxidant and Anti-obesity Effects of Herbal Complex Extract (한방복합추출물의 항산화 및 항비만 효과)

  • Shim, Jae-Won;Lee, Seung-Ju;Kim, Hye Kyung;Choi, Yun-Sik;Jang, Young-Ah
    • Journal of Life Science
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    • v.32 no.7
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    • pp.523-531
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    • 2022
  • The purpose of this study was to evaluate the effect of antioxidant and anti-adipogenic activities in ethanol extracts from herb mixture (Ephedra sinica, Atractylodes lance, Gypsum fibrosum, and Theobroma cacao). DPPH, ABTS+ radical and xanthine oxidase scavenging activities were measured for antioxidant activity. Extracts of the herb mixture had 75.0, 100.8, and 79.5% scavenging activities at 1,000 ㎍/ml concentration, respectively. We investigated the inhibition of adipogenesis and adipocyte differentiation with an extract of an herb mixture in 3T3-L1 preadipocytes. An extract from the herb mixture at concentrations between 0 and 50 ㎍/ml did not affect 3T3-L1 cell viability. Treatment with herb mixture extracts of 25, 50, and 75 ㎍/ml in 3T3-L1 preadipocytes inhibited lipid accumulation in a dose-dependent manner. As a result of a Western blot experiment, it was shown that the herb mixture inhibited the differentiation transcription factors, PPARγ and C/EBPa, by 44.2 and 77.6%, respectively, at a concentration of 75 ㎍/ml in MDI-induced differentiated 3T3-L1 cells. As a result of RT-PCR, the gene expression of C/EBPa, SREBP-1c, and PPARγ was significantly inhibited by 43.4%, 59.6%, and 55.3%, respectively, at the concentration of 75 ㎍/ml of the herb mixture compared with the MDI-treated group. In addition, the expression of fatty acid synthase (FAS), a fatty acid synthesis regulator, was suppressed. These results can be applied to develop a functional food for anti-obesity with a herb mixture.

Effects of Alisma canaliculatum Extract in Pacemaker Potential of Intestinal Interstitial cells of Cajal in mice (생쥐 소장 및 대장 카할세포의 자발적 탈분극에서 택사의 효과에 관한 비교연구)

  • Kwon, Hyo Eun;Park, Dong Suk;Kim, Jeong Nam;Kim, Byung Joo
    • Herbal Formula Science
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    • v.30 no.2
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    • pp.37-44
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    • 2022
  • Objectives : The purpose of this study was to examine the effects of Alisma canaliculatum Extract (ACE) on pacemaker potentials of small and large intestinal interstitial Cells of Cajal (ICC) in mice. Methods : We used enzymatic digestions to dissociate the ICC in the small and large intestine in mice. The whole-cell patch-clamp method was used to record pacemaker potentials in ICC. Results : 1. The ICC generated the pacemaker potentials in small intestine in mice. ACE (0.1-1mg/ml) induced membrane depolarization and decreased frequency with concentration-dependent manners. 2. Pretreatment with a Ca2+ free solution, Na+ 5 mM solution or 2-APB, a nonselective cation channel blocker, stopped the small intestinal ICC pacemaker potentials. In the case of Ca2+-free solution, Na+ 5 mM solution or 2-APB, ACE had no effects on the membrane depolarizations in small intestinal ICC. 3. The ICC generated the pacemaker potentials in large intestine in mice. Membrane depolarization appears regularly in the small intestine, but irregularly in the large intestine. ACE induced membrane depolarization (0.1-1mg/ml) and increased frequency (0.1-0.5mg/ml). 4. Pretreatment with a Ca2+ free solution, Na+ 5 mM solution or 2-APB, stopped the large intestinal ICC pacemaker potentials. In the case of Ca2+-free solution, Na+ 5 mM solution or 2-APB, ACE depolarized the membrane depolarizations in large intestinal ICC. 5. In mice, intestinal transit rate (ITR) values were dose-dependently decreased by the intragastric administration of ACE. Conclusions : These results suggest that ACE can regulate the pacemaker activity of ICC and the reaction by ACE is different from the small and large intestinal ICC, and the control of the intestinal motion by ACE may be caused by many complex processes.