• Title/Summary/Keyword: Molecular aggregation

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Inhibitory effects of artemether on thrombus formation via regulation of cyclic nucleotides in collagen-induced platelets (콜라겐-유도의 혈소판에서 사이클릭 뉴클레오티드의 조절을 통한 Artemether의 항혈전 효과)

  • Chang-Eun Park;Dong-Ha Lee
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.239-245
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    • 2022
  • Although normal activation of platelets is important in the process of hemostasis, excessive or abnormal activation of platelets can lead to cardiovascular diseases. Therefore, the discovery of novel substances capable of regulating or inhibiting platelet activation may be helpful in the prevention and treatment of cardiovascular diseases. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. This study investigated the effects of artemether on platelet activation and thrombus formation induced by collagen. As a result, cAMP level was significantly increased by artemether, and VASP and IP3R, substrates of cAMP-dependent kinase, were phosphorylated. IP3R phosphorylation by Artemether inhibited Ca2+ recruitment into the cytoplasm, and phosphorylated VASP inhibited fibrinogen binding by inactivating αIIb/β3 located on the platelet membrane. Consequently, artemether inhibited thrombin-induced fibrin clot formation. Therefore, we propose that artemether can act as an effective prophylactic and therapeutic agent for cardiovascular diseases caused by excessive platelet activation and thrombus formation.

EFFECTS OF SIGNAL TRANSDUCTION PATHWAY IN THE RAS-INDUCED CELLULAR TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (인체 상피세포에서 ras-종양유전자의 발암화가 신호 전달 기작에 미치는 영향)

  • Jang, Do-Geun;Byeon, Ki-Jeong;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.3
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    • pp.254-261
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    • 2000
  • The present study has attempted to look into the mechanism of ras-induced carcinogenesis in a human epithelial cell system. Human epithelial cells immortalized with Ad12-SV40 hybrid virus were used to assess carcinogenic potential of the ras-oncogene. Cells transfected with pSV2-ras showed characteristics of cellular transformation. The transformation parameters such as cell density, soft-agar colony formation, and cell aggregation were significantly increased in the cells expressing ras oncoprotein. In addition, the duration required for the appearance of foci was shortened in the ras-transfected cells. Consistent with other reports, our results demonstrated an evidence that the ras-oncogene induced the cellular transformation of human epithelial cell system. When a high concentration of glucocorticoid was added into the media, transformation process was accelerated. It is speculated that glucocorticoid may provide an advantageous environment for the proliferation of the transformed cells. The induction of the intracellular free calcium concentrations following agonist treatment was significantly lower in the transformed cells than in the control cells. These effects were more manifested in the presence of extracellular cacium, indicating that the transformation process may alter the influx pathway of extracellular calcium. The induction of $IP_3$ following agonist treatment was also lower in the transformed cells than in the control cells. Thus, it is suggested that phospholipase C-coupled pathway was down-regulated in the process of the ras-induced transformation. While the levels of $TGF-{\beta}_1$ and PAI-2 mRNAs were decreased, the level of fibronectin mRNA was increased. The results indicate that mechanism of the ras-induced transformation may be associated with the altered expressions of growth regulatory factors. The present study demonstrates an evidence that the ras-induced cellular transformation may be associated with alteration of signal transduction and growth regulatory factors. The study will contribute to improve the understanding of molecular mechanism of epithelium-derived cancers including oral cancer.

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Pharmaceutical Characteristics of Korean Lumbricus rubellus Lumbrokinase (한국산 지렁이[Lumbricus rubellus]에서 분리한 Lumbrokinase의 약리학적 특성)

  • 조일환;이철규;임헌길;이형환
    • KSBB Journal
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    • v.19 no.4
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    • pp.274-283
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    • 2004
  • Six lumbrokinase (LK) fractions from Lumbricus rubellus lysates were purified by a series of column chromatographies. The molecular weights of the six LK fractions appeared to range from 24.6 to 33.1 kDa. In the experimental model of rat venous thrombosis, the thrombus weight and PAI activity decreased significantly when the LK was administered orally. However, the activities of APTT, PT and plasmin showed a significant increase. The aggregation of rat platelets pretreated with various LK doses was inhibited by thrombin, and the MDA generation decreased. The rat thoracic aorta and mesentric arteries contracted with phenylephrine relaxed due to the treatment of the LK fractions. These results suggest that the fibrinolytic effects of LK were mediated not only by proteolytic activity, but also by the inhibition of platelet agregation and the relaxation of blood vessels. It is concluded that the LK may be useful as a hemolytic agent for treatment of fibrin clot.

Effect of High Dose ${\gamma}-irradiation$ on the Physicochemical Properties of Shell Eggs during Storage (고선량 감마선 조사가 신선란의 저장 중 이화학적 특성에 미치는 영향)

  • Moon, Sun-Ae;Song, Kyung-Bin
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.260-265
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    • 2000
  • To determine the quality change of the irradiated eggs during storage, fresh shell eggs were irradiated using $^{60}Co$ at 0, 1, 5, 10, 30 kGy and stored for 30 days. The york index, color, pH, viscosity, egg weight, and SDS-PAGE profile of the irradiated eggs were examined. During storage, york index values of the irradiated eggs and the control were decreased and the increase of dose decreased yolk index. However, the yolk index values were increased temporarily at 10 kGy and 30 kGy. The yolk color had a bright yellow with increases in dose level and there was no significant change during storage. The albumen viscosities were decreased with increases in dosage and were decreased during storage. Also, the albumen pH values of the irradiated eggs were higher than that of the control and were increased during storage. The weight losses of eggs were increased during storage and there were no significant changes by dose level. SDS-PAGE profile of the egg white proteins of the shell eggs showed the change in molecular weight distribution and had aggregation pattern as well as degradation. CD and fluorescence spectroscopy study showed changes in the secondary and tertiary structure of egg white proteins by ${\gamma}-irradiation$. Therefore, this study clearly indicates that irradiation dose of eggs should be appropriate to prevent the loss of egg qualities.

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Solid-Phase Refolding of Poly-Lysine fusion Protein of hEGF and Angiogenin (Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘)

  • Park, Sang-Joong;Ryu, Kang;Suh, Chang-Woo;Chai, Young-Gyu;Kwon, Oh-Byung;Park, Seung-Kook;Lee, Eun-Kyu
    • KSBB Journal
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    • v.17 no.2
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    • pp.153-157
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.

Stability increase in the activity of tolaasin inhibitors under reducing conditions (환원 조건에서 톨라신 저해 물질 활성의 안정성 증가)

  • Yun, Yeong-Bae;Kim, Min-Hee;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.60 no.4
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    • pp.351-355
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    • 2017
  • Tolaasin, peptide toxin produced by Pseudomonas tolaasii, causes a brown blotch disease on the cultivated mushrooms. Tolaasin peptides form membrane pores and disrupt cellular membrane structure. Molecular actions of tolaasin consist of the aggregation of peptide molecules, binding to the cell membrane, and formation of membrane pores. Therefore, the inhibitions of any of these actions are able to suppress the blotch disease. We have isolated and identified several tolaasin inhibitors (named tolaasin inhibitory factors, TIF) from food additives. TIFs were able to suppress the blotch-formation by the pathogen inoculated to the mushrooms. In this study, TIFs were incubated under various conditions and their activities for the inhibition of tolaasin-induced hemolytic activity were investigated. Since TIFs are unsaturated carbon compounds, they were sensitive to the air exposure and light irradiation. In the anaerobic conditions, TIFs were stable and their activities were decreased by 10% for three months. However, near 90% of TIF activity was suppressed by two weeks in the presence of air and sun light. Temperature did not show any significant effects on the activity of TIF, since storages at 5, 25, $45^{\circ}C$ did not show any difference. Therefore, for the stable storage of TIF compounds, container should be designed to be dark and air-tight.

Expression of HSP70 Immunoreactivity in EPO Treated Rat Kidney (콩팥에서 Erythropoietin 투여로 인한 HSP70의 발현 변화)

  • Jung, Ju-Young;Kim, Jin
    • Applied Microscopy
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    • v.37 no.3
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    • pp.167-174
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    • 2007
  • Heat shock protein (HSP) 70 functions as a molecular chaperon and reduces stress-induced denaturation and aggregation of intracellular proteins. Erythropoietin (EPO) plays an important role during acute renal failure repair process by rapidly correcting anemia and enhancing renal tubular regeneration. The purpose of this study was to examine the effect of EPO treatment on renal HSP70 expression. Male Sprague-Dawley rats were injected rHUEPO. Kidney were preserved by in vivo perfusion with paraformaldehyde-lysine-periodate (PLP) and processed for immunohistochemistry and electron microscopy. In control kidney, HSP70 was expressed in the cortex, outer medulla and inner medulla. Especially, HSP immunoreactiviy was mainly founded in descending thin limb of outer medulla and inner medullary collecting duct. In EPO treated kidney, HSP70 expression markedly increased in the descending thin limb of outer medulla and newly detected in cortical collecting duct. Electron microscopy showed the presence of HSP immunoreactivity on the intracelluar vesicles and Golgi complex of descending thin limb and cortical collecting duct. These findings suggest that EPO treatment leads to new production of HSP70 in renal tubular cells, and induction of HSP70 by rHuEPO is causally related to protective function.

Effect of Amine Functional Group on Removal Rate Selectivity between Copper and Tantalum-nitride Film in Chemical Mechanical Polishing

  • Cui, Hao;Hwang, Hee-Sub;Park, Jin-Hyung;Paik, Ungyu;Park, Jea-Gun
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.546-546
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    • 2008
  • Copper (Cu) Chemical mechanical polishing (CMP) has been an essential process for Cu wifing of DRAM and NAND flash memory beyond 45nm. Copper has been employed as ideal material for interconnect and metal line due to the low resistivity and high resistant to electro-migration. Damascene process is currently used in conjunction with CMP in the fabrication of multi-level copper interconnects for advanced logic and memory devices. Cu CMP involves removal of material by the combination of chemical and mechanical action. Chemicals in slurry aid in material removal by modifying the surface film while abrasion between the particles, pad, and the modified film facilitates mechanical removal. In our research, we emphasized on the role of chemical effect of slurry on Cu CMP, especially on the effect of amine functional group on removal rate selectivity between Cu and Tantalum-nitride (TaN) film. We investigated the two different kinds of complexing agent both with amine functional group. On the one hand, Polyacrylamide as a polymer affected the stability of abrasive, viscosity of slurry and the corrosion current of copper film especially at high concentration. At higher concentration, the aggregation of abrasive particles was suppressed by the steric effect of PAM, thus showed higher fraction of small particle distribution. It also showed a fluctuation behavior of the viscosity of slurry at high shear rate due to transformation of polymer chain. Also, because of forming thick passivation layer on the surface of Cu film, the diffusion of oxidant to the Cu surface was inhibited; therefore, the corrosion current with 0.7wt% PAM was smaller than that without PAM. the polishing rate of Cu film slightly increased up to 0.3wt%, then decreased with increasing of PAM concentration. On the contrary, the polishing rate of TaN film was strongly suppressed and saturated with increasing of PAM concentration at 0.3wt%. We also studied the electrostatic interaction between abrasive particle and Cu/TaN film with different PAM concentration. On the other hand, amino-methyl-propanol (AMP) as a single molecule does not affect the stability, rheological and corrosion behavior of the slurry as the polymer PAM. The polishing behavior of TaN film and selectivity with AMP appeared the similar trend to the slurry with PAM. The polishing behavior of Cu film with AMP, however, was quite different with that of PAM. We assume this difference was originated from different compactness of surface passivation layer on the Cu film under the same concentration due to the different molecular weight of PAM and AMP.

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Spatial Distribution Pattern of the Populations of Cephalanthera erecta at Mt. Ahop in Busan (부산시 아홉산의 은난초 집단의 공간적 분포 양상)

  • Huh, Man Kyu
    • Journal of Life Science
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    • v.26 no.8
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    • pp.881-886
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    • 2016
  • Cephalanthera erecta (Thunb. ex, Murray) is an herbaceous and a member of the genus Cephalanthera in the family Orchidaceae. The species is an herbaceous and has reputed Chinese medicinal value. It has been investigated the population density and spatial distribution of this species at Mt. Ahop in Korea during 2015. The spatial pattern of C. erecta was analyzed according to several patchiness indexes, population uniformity or aggregation under different sizes of plots by dispersion indices, and spatial autocorrelation. The mean crowding (M*) and patchiness index (PAI) showed positive values except one small plot (2 m × 2 m). Most natural individuals of C. erecta for plots were not uniformly distributed in the forest community. The small plots (2 m × 2 m, to 8 m × 16 m) of C. erecta were uniformly distributed in the forest community and large plots (16 m × 16 m and 16 m × 32 m) were aggregately distributed. Significant aggregations by Moran's I of C. erecta were partially observed within IV classes (12 m). Dissimilarity among pairs of individuals could found by more than 18.0 m. In conclusion, the geographic distribution of C. erecta is not even with varying degrees of size of plots and human activities give rise to density effects in the plots at Mt. Ahop in Korea.

Geochemical Characteristics of Granodiorite and Arenaceous Sedimentary Rocks in Chon-Ashuu Area, Kyrgyzstan (키르키스스탄 촌아슈 지역 화강섬록암질암 및 사질원 퇴적암의 지화학적 특징)

  • Kim, Soo-Young;Chi, Sei-Jung;Park, Sung-Won
    • Economic and Environmental Geology
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    • v.44 no.4
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    • pp.273-288
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    • 2011
  • Chon-Ashuu copper mining claim area is located, in terms of the geotectonic setting, in the northern part of the suture line which is bounded with the marginal part of Issik-kul micro-continent on the southern part of North Tien-Shan terrane. The geological blocks of Chon-Ashuu districts belong to the southern tip of Kazakhstan orocline. The rock formation of this area are composed of the continental crust or/and arc collage and the paleo-continental fragments-accretionary wedge complex of pre-Altaid orogenic materials. ASI(Alumina Saturation Index) of Paleozoic plutonic rocks in Chon-Ashuu area belong to the peraluminous and metaluminous rocks which were generated from fractional crystallization of Island and volcanic arc crusts in syn-post collisional plate. The geology of the ChonAshuu area consists of upper Proterozoic and Paleozoic rock formations. According to Harker variation diagrams for Chon-Ashuu arenaceous sedimentary rocks, the silty sandstone of Chon-Ashuu area showing the mineralogical immaturity were derived from Island arc or the marginal environments of active continent in Cambro-Carboniferous period. Numerous intrusive rocks of Chon-Ashuu area are distributed along north east trending tectonic structures and are bounded on four sides by the conjugate pattern. The most common type of the plutonic rocks are granodiorite and monzodiorite. According to the molecular normative An-Ab-Or composition (Barker, 1979), the plutonic rocks in Chon-Ashuu area are classified into tonalite - trondhjemite - granodiorite (TTG) series which are an aggregation of rocks which is the country rock of copper mineralization, that are formed by melting of hydrous mafic crust at high pressure.