• Title/Summary/Keyword: protein transport

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A study of the lipoprotein lipase inhibitory mechanism of Poncirus trifoliata water extracts (탱자 (Poncirus trifoliata)의 lipoprotein lipase 억제메커니즘)

  • Lee, Sung Mee;Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Tae Woo;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.48 no.1
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    • pp.9-18
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    • 2015
  • Purpose: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. Methods: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein ($C/EBP{\beta}$). Results: The total polyphenol and flavonoid content of PF-W was $52.15{\pm}4.02$ and $6.56{\pm}0.47mg/g$, respectively. PF-W treatment decreased LPL content in media to $58{\pm}5%$ of that in control adipocyte media, and increased LPL content to $117{\pm}3.5%$ of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of $C/EBP{\beta}$, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. Conclusion: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.

Dependence of Sub-Cellular Activities of the Blooming and Harmful Dinoflagellate Cochlodinium Polykrikoides on Temperature (수온에 따른 유해성 Cochlodinium polykrikoides 적조생물의 세포생리 변화)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1194-1201
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    • 2008
  • Water temperature-dependent fluctuations of biochemical and molecular activities in the harmful dinoflagellate, Cochlodinium polykrikoides were studied. In terms of genomic DNA concentration, a similar value of 0.6 was observed at $12^{\circ}C$ and $15^{\circ}C$. However, DNA significantly increased beyond $18^{\circ}C$ (p<0.05), to a maximum of 1.8 at $24^{\circ}C$. DNA concentration significantly decreased to 0.6. The concentrations of RNA and total protein were likely at their highest values of 1.7 and 0.07 ${\mu}g$ $ml^{-1}$ at $24^{\circ}C$, respectively. RNA and total protein concentrations began to increase at $15^{\circ}C$. Oxygen availability between lower and higher temperatures was significantly different and increased from $18^{\circ}C$ according to light intensity, regardless of wavelengths (p<0.05). At $24^{\circ}C$, the highest value of the maximum electron transport rate ($ETR_{max}$), ranging from 537.9 (Ch 1) to 602.5 ${\mu}mol$ electrons $g^{-1}$ Chl ${\alpha}s^{-1}$ (Ch 4), was also apparent. Nitrate reductase (NR) and ATPase activities were at their highest values of 0.11 ${\mu}mol$ $NO_{2}^{-}$ ${\mu}g^{-1}$ Chl ${\alpha}h^{-1}$ and 0.78 pmol 100 $mg^{-1}$ at $24^{\circ}C$, respectively. In an analysis of CHN, the concentration of C and N also significantly increased (p<0.05). Most of the measurements for the cellular activities at $27^{\circ}C$, however, were less than at $24^{\circ}C$. These results suggest that the sub-cellular activities of C. polykrikoides are sensitive to changes in water temperature. It may be desirable to estimate at $18^{\circ}C$ the initiation of the massive blooming development of C. polykrikoides. In nature, it will be very difficult to maintain the massive blooms beyond $24^{\circ}C$ because of a possibly significant decrease in molecular activity of C. polykrikoides.

Functional implications of gene expression analysis from rice tonoplast intrinsic proteins during seed germination and development (벼 종자에서 액포막 aquaporin (tonoplast intrinsic protein) 유전자의 발현과 기능)

  • Huh, Sun-Mi;Lee, In-Sook;Kim, Beom-Gi;Shin, Young-Seop;Lee, Gang-Seop;Kim, Dool-Yi;Byun, Myung-Ok;Kim, Dong-Hern;Yoon, In-Sun
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.517-528
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    • 2010
  • Rice seed maturation and germination involve drastic changes in water and nutrient transport, in which tonoplast aquaporins may play an important role. In the present study, gene expression profiles of 10 tonoplast intrinsic proteins (TIP) from rice were investigated by RT-PCR during seed development and germination. OsTIP3;1 and OsTIP3;2 were specifically expressed in mature seeds. Their transcript level rapidly decreased after onset of seed germination and gene expression was induced by ABA treatment. In contrast, expression of OsTIP2;1 and OsTIP4;3 was not seed specific as transcripts were found in vegetative tissues as well. Their respective transcript levels decreased at an early stage of seed development, whereas they increased at a later stage of seed germination and elongation of embryonic roots and shoots. When seed germination was inhibited by various stress conditions and ABA, expression of OsTIP2;1 and OsTIP4;3 was completely suppressed. In contrast, the expression level of OsTIP2;2 rapidly increased after seed imbibition and the transcript level was maintained under conditions inhibiting seed germination. These results implicate that tissue specific and developmental transcriptional regulation of OsTIPs in rice seeds depends on their specific function. In addition, OsTIPs can be discriminated by different potential phosphorylation and methylation sites in their protein structures. OsTIP3;1 and OsTIP3;2 possess unique phosphorylation signatures at their N-terminal domain, loop B and loop E, respectively. OsTIP2;1 and OsTIP4;3 have a potential methylation site at their Nterminal domain. This suggests that activity of specific tonoplast aquaporins may be regulated by post-translational modification as well as by transcriptional control.

Cellular Energy Allocation of a Marine Polychaete Species (Perinereis aibuhitensis) Exposed to Dissolving Carbon Dioxide in Seawater (해수 중 용존 이산화탄소 농도 증가가 두토막눈썹참갯지렁이(Perinereis aibuhitensis)의 세포내 에너지 할당에 미치는 영향)

  • Moon, Seong-Dae;Lee, Ji-Hye;Sung, Chan-Gyoung;Choi, Tae Seob;Lee, Kyu-Tae;Lee, Jung-Suk;Kang, Seong-Gil
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.16 no.1
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    • pp.9-16
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    • 2013
  • An experiment was conducted to evaluate the biochemical adverse effect of increased carbon dioxide in seawater on marine polychaete, Perinereis aibuhitensis. We measured the available energy reserves, Ea (total carbohydrate, protein, and lipid content) and the energy consumption, Ec (electron transport activity) of Perinereis aibuhitensis exposed for 7-d to a range of $CO_2$ concentration such as 0.39 (control =390 ppmv), 3.03 (=3,030 ppmv), 10.3 (=10,300 ppmv), and 30.1 (=30,100 ppmv) $CO_2$ mM, respectively. The cellular energy allocation (CEA) methodology was used to assess the adverse effects of toxic stress on the energy budget of the test organisms. The results of a decrease in CEA effect of increased carbon dioxide in seawater from all individual in Ea and Ec. Increase of carbon dioxide reduced pH in seawater, significantly. The chemical changes in sea- water caused by increasing $pCO_2$ might cause stresses to test organisms and changes in the cellular energy allocations. Results of this study can be used to understand the possible influence of $CO_2$ concentration increased by the leakage from sub-sea bed storage sites as well as fossil fuel combustion on marine organisms.

Detection of Multidrug Resistance Using Molecular Nuclear Technique (분자핵의학 기법을 이용한 다약제내성 진단)

  • Lee, Jae-Tae;Ahn, Byeong-Cheol
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.2
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    • pp.180-189
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    • 2004
  • Although the outcome of cancer patients after cytotoxic chemotherapy is related diverse mechanisms, multidrug resistance (MDR) for chemotherapeutic drugs due to cellular P-glycoprotein (Pgp) or multidrug-resistance associated protein (MRP) is most important factor in the chemotherapy failure to cancer. A large number of pharmacologic compounds, including verapamil, quinidine, tamoxifen, cyclosporin A and quinolone derivatives have been reported to overcome MDR. Single photon emission computed tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transporter. $^{99m}Tc$-MIBI and other $^{99m}Tc$-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of PgP-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with $^{11}C$ have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and $N-[^{11}C]acetyl-leukotriene$ E4 provides an opportunity to study MRP function non-invasively in vivo. SPECT and PET pharmaceuticals have successfully used to evaluate pharmacologic effects of MDR modulators. Imaging of MDR and reversal of MDR with bioluminescence in a living animal is also evaluated for future clinical trial. We have described recent advances in molecular imaging of MDR and reviewed recent publications regarding feasibility of SPECT and PET imaging to study the functionality of MDR transporters in vivo.

Changes of Stress Response and Physiological Metabolic Activity of Flounder, Paralichthys olivaceus Following to Food Deprivation and Slow Temperature Descending (먹이제한과 단기 수온하강 조건에서 넙치의 스트레스 반응과 생리학적 대사활성 변화)

  • Myeong, Jeong-In;Kang, Duk-Young;Kim, Hyo-Chan;Lee, Jeong-Ho;Noh, Jae-Koo;Kim, Hyun-Chul
    • Korean Journal of Ichthyology
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    • v.23 no.2
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    • pp.87-94
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    • 2011
  • To find the preliminary environmental conditions for a short-time transport of living olive flounder, Paralichthys olivaceus, the stress response and physiological metabolic activity of the cultured fish to feed deprivation and slow temperature descending ($15.8^{\circ}C{\rightarrow}13.3^{\circ}C$) were monitored for 8 days. The monitored variables were the plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose (GLU), total protein (TP), electrolytes ($Na^+$, $K^+$, $Cl^-$) and thyroid hormones ($TT_4$, $TT_3$, $FT_4$ and $FT_3$). In food deprivation experiment for 8 days, we did not find any statistical change of level in AST, ALT and electrolytes ($Na^+$, $K^+$, $Cl^-$), but found a significant decrease in TP and GLU. In thyroid hormones, the levels of four hormones in plasma were all showing a tendency to decrease. Especially, $FT_4$ and $TT_3$ were significantly decreased, indicating a withering of physiologic activity. In the temperature test, although no any significant change in AST, TP and electrolytes ($Na^+$, $K^+$, $Cl^-$), we observed a significant decrease of ALT and GLU following to temperature descending from $15.8^{\circ}C$ to $13.3^{\circ}C$ (P<0.05). In the levels of thyroid hormones, any significant change was not observed for experimental period. We conclude that the stress response and physiological activity of olive flounder were more influenced by feed deprivation than slow temperature descending at a transport of living fish, and plasma GLU appears to be sensitive factor to physiological metabolic activity, indicating that it could be used as a monitering mark or index for a health inspection of the fish.

Dietary L-carnitine Influences Broiler Thigh Yield

  • Kidd, M.T.;Gilbert, J.;Corzo, A.;Page, C.;Virden, W.S.;Woodworth, J.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.5
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    • pp.681-685
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    • 2009
  • L-carnitine promotes mitochondrial ${\beta}$-oxidation of long chain fatty acids and their subsequent transport across the inner mitochondrial membrane. Although the role of L-carnitine in fatty acid metabolism has been extensively studied, its role in live performance and carcass responses of commercial broilers is less understood. The objective of this research was to determine if Lcarnitine fed at various levels in diets differing in CP and amino acids impacted on live performance and carcass characteristics of commercial broilers. Two floor pen experiments were conducted to assess the effect of dietary L-carnitine in grower diets. In Exp. 1, Ross${\times}$Hubbard Ultra Yield broilers were placed in 48 floor pens (12 birds/pen) and fed common diets to d 14. A two (0 or 50 ppm Lcarnitine) by three (173, 187, and 202 g/kg CP) factorial arrangement of treatments was employed from 15 to 35 d of age (8 replications/treatment). An interaction (p<0.05) in carcass yield indicated that increasing CP (187 g/kg) resulted in improved yield in the presence of L-carnitine. Increasing CP from 173 to 202 g/kg increased (p<0.05) BW gain and decreased (p<0.05) feed conversion and percentage abdominal fat. Feeding dietary L-carnitine increased back-half carcass yield which was attributable to an increase (p<0.05) in thigh, but not drumstick, yield relative to carcass. In Exp. 2, $Ross{\times}Ross$ 708 broilers were fed common diets until 29 d. From 30 to 42 d of age, birds were fed one of seven diets: i) 200 g/kg CP, 0 ppm L-carnitine; ii) 200 g/kg CP, 40 ppm L-carnitine; iii) 180 g/kg CP, 0 ppm L-carnitine; iv) 180 g/kg CP, 10 ppm L-carnitine; v) 180 g/kg CP, 20 ppm L-carnitine; vi) 180 g/kg CP, 30 ppm L-carnitine; and vii) 180 g/kg CP, 40 ppm L-carnitine (6 replications of 12 birds each). BW gain, feed conversion, mortality (30 to 42 d), and carcass traits (42 d) were measured on all birds by pen. There were no treatment differences (p<0.05). However, the addition of 40 ppm L-carnitine in the 200 g CP/kg diet increased (p = 0.06) thigh yields relative to BW in comparison to birds fed diets without L-carnitine, which was further confirmed via a contrast analysis (0 vs. 40 ppm L-carnitine in the 200 and 180 g CP/kg diets; p<0.05). These results indicated that dietary L-carnitine may heighten metabolism in dark meat of commercial broilers resulting in increased relative thigh tissue accretion without compromising breast accretion.

Identification of Differentially Expressed Genes Related to Intramuscular Fat Development in the Early and Late Fattening Stages of Hanwoo Steers

  • Lee, Seung-Hwan;Park, Eung-Woo;Cho, Yong-Min;Kim, Sung-Kon;Lee, Jun-Heon;Jeon, Jin-Tae;Lee, Chang-Soo;Im, Seok-Ki;Oh, Sung-Jong;Thompson, J.M.;Yoon, Du-Hak
    • BMB Reports
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    • v.40 no.5
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    • pp.757-764
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    • 2007
  • Marbling of cattle meat is dependent on the coordinated expression of multiple genes. Cattle dramatically increase their intramuscular fat content in the longissimus dorsi muscle between 12 and 27 months of age. We used the annealing control primer (ACP)-differential display RT-PCR method to identify differentially expressed genes (DEGs) that may participate in the development of intramuscular fat between early (12 months old) and late fattening stages (27 months old). Using 20 arbitrary ACP primers, we identified and sequenced 14 DEGs. BLAST searches revealed that expression of the MDH, PI4-K, ferritin, ICER, NID-2, WDNMI, telethonin, filamin, and desmin (DES) genes increased while that of GAPD, COP VII, ACTA1, CamK II, and nebulin decreased during the late fattening stage. The results of functional categorization using the Gene Ontology database for 14 known genes indicated that MDH, GAPD, and COP VII are involved in metabolic pathways such as glycolysis and the TCA cycle, whereas telethonin, filamin, nebulin, desmin, and ACTA1 contribute to the muscle contractile apparatus, and PI4-K, CamK II, and ICER have roles in signal transduction pathways regulated by growth factor or hormones. The final three genes, NID-2, WDNMI, and ferritin, are involved in iron transport and extracellular protein inhibition. The expression patterns were confirmed for seven genes (MDH, PI4-K, ferritin, ICER, nebulin, WDNMI, and telethonin) using real-time PCR. We found that the novel transcription repressor ICER gene was highly expressed in the late fattening stage and during bovine preadipocyte differentiation. This information may be helpful in selecting candidate genes that participate in intramuscular fat development in cattle.

Inhibitory mechanism of α1-adrenergic stimulation on the release of thyroxine in mouse thyroids (Mouse 갑상선에서 α1-adrenoceptor 자극에 의한 thyroxine 유리 억제기전)

  • Kang, Hyung-sub;Kim, Song-kyu;Kang, Chang-won;Kim, Jin-sang;Lee, Ho-il
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.712-719
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    • 1998
  • Thyroid function is mainly regulated through cAMP and phophatidylinositol, and it is well known that TSH-stimulated thyroxine ($T_4$) release is inhibited by catecholamine from mouse thyroids via the ${\alpha}_1$-adrenoceptor stimulation. Previous study has established that the inhibition of $T_4$ release by ${\alpha}_1$-adrenoceptor stimulation results in activated protein kinase C (PKC). The purpose of this study was to determine if ion transport systems are involved in the inhibition of $T_4$ release elicited by ${\alpha}_1$-adrenergic agonist in mouse thyroids. TSH-, IBMX- and cAMP analogue-stimulated $T_4$ release were significantly inhibited by methoxamine, R59022 (diacylglycerol kinase inhibitor), and MDL (adenylate cyclase inhibitor). TSH-stimulated $T_4$ release could be inhibited by Bay K 8644 and cyclopiazoic acid, but not by verapamil and tetrodotoxin. The addition of nifedipine ($Ca^{2+}$ channel blocker), tetrodotoxin and lidocaine ($Na^+$ channel blockers), but not amiloride (EIPA) and ryanodine, completely blocked the inhibitory effects of methoxamine on $T_4$ release. TSH-stimulated $T_4$ release was also inhibited by benzamil ($Na^+-Ca^{2+}$ exchange inhibitor). TSH-, IBMX- and cAMP-stimulated $T_4$ release were inhibited by methoxamine or R59022, these effects were reversed by nifedipine. but not by verapamil. Furthermore, nifedipine reversed the inhibitory effects of benzamil and R59022 on TSH-stimulated $T_4$ release. These data suggest that the observed ${\alpha}_1$-adrenoceptor-mediated inhibition of $T_4$ release in mouse thyroids is the result of an increase in intracellular $Na^+$ or $Ca^{2+}$ effected via activation of fast $Na^+$ or nifedipine-sensitive $Ca^{2+}$ channels, and that $Na^+-Ca^{2+}$ exchange may play an important role in reducing thyroid hormone by increasing intracellular $Ca^{2+}$.

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Identification of Blood Cells and Their Physiological Functions in the Scallop, Patinopecten yessoensis (가리비, Patinopecten yessoensis 혈구의 형태학적 동정과 기능)

  • CHANG Young Jin
    • Journal of Aquaculture
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    • v.6 no.1
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    • pp.1-12
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    • 1993
  • Identification of blood cells and their physological functions in the cultured scallop, Patinopecten yessoensis collected from Abashiri Bay, Hokkaido, Japan were studied by electron microscopic structure and histological observations. The physiological function of each blood cell type was studied on the basis of its cytological structure, the lysosome in the blood cell and its phagocytosis. The blood cells were classified as Type I, Type II and Type III. The morphological characteristics of each blood cell type are as follows ; Type I : The cell is oval shaped and its cytoplasm contains comparatively low electron dense materials. The oval nucleus is sometimes ramified into two nuclei. Lumps of tubular smooth endoplasmic reticula and vacuoles are distributed near the nucleus. Type II : The cell appears long and oval shaped, and its cytoplasm contains high electron dense materials. The oval nucleus does not ramify, and large numbers of sac-like smooth endoplasmic reticula and free ribosomes are developed around the nucleus. No vacuoles exist in the cytoplasm Type 1II : The cell is round in shape and the electron density of the cytoplasm is the highest among the three types of cells because of large quantities of rough­surfaced endoplasmic reticula and no vacuoles. Particularly, the nucleus reveals a wheel-like shape owing to lumps of tuberous chromatin. The cells of Type I and II seem to have the role of carrying out phagocytosis on either foreign materials such as bacteria or endogenous old cells and the transport of nutritive materials. The type III cell, which has not been found in any bivalve species of non Pectinidae, may be said to have the function of production and the secretion of protein related to some humoral defense materials.

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