• Title/Summary/Keyword: density gradient

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Caffeine Indirectly Activates Ca2+-ATPases in the Vesicles of Cardiac Junctional Sarcoplasmic Reticulum

  • Kim, Young-Kee;Cho, Hyoung-Jin;Kim, Hae-Won
    • BMB Reports
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    • v.29 no.1
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    • pp.22-26
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    • 1996
  • Agents that activate or inhibit the $Ca^{2+}$ release channel in cardiac sarcoplasmic reticulum (SR) were tested for their abilities to affect the activity of the SR $Ca^{2+}$-ATPase. Vesicles of junctional SR (heavy SR, HSR) from terminal cisternae were prepared from porcine cardiac muscle by density gradient centrifugation. The steady-state activity of $Ca^{2+}$-ATPases in intact HSR vesicles was/$347{\pm}5\;nmol/min{\cdot}mg$ protein (${\pm}$ SD). When the HSR vesicles were made leaky, the activity was increased to $415{\pm}5\;nmol/min{\cdot}mg$ protein. This increase is probably due to the uncoupling of HSR vesicles. Caffeine (10 mM), an agonist of the SR $Ca^{2+}$ release channel, increased $Ca^{2+}$-ATPase activity in the intact HSR vesicle preparation to $394{\pm}30\;nmol/min{\cdot}mg$ protein. However, caffeine had no significant effect in the leaky vesicle preparation and in the purified $Ca^{2+}$-ATPase preparation. The effect of caffeine on SR $Ca^{2+}$-ATPase was investigated at various concentrations of $Ca^{2+}$. Caffeine increased the pump activity over the whole range of $Ca^{2+}$ concentrations, from $1\;{\mu}M$ to $250\;{\mu}M$, in the intact HSR vesicles. When the SR $Ca^{2+}$-ATPase was inhibited by thapsigargin, no caffeine effect was observed. These results imply that the caffeine effect requires the intact vesicles and that the increase in $Ca^{2+}$-ATPase activity is not due to a direct interaction of caffeine with the enzyme. We propose that the activity of SR $Ca^{2+}$-ATPase is linked indirectly to the activity of the $Ca^{2+}$ release channel (ryanodine receptor) and may depend upon the amount of $Ca^{2+}$ released by the channels.

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An experimental study on the local velocity acceleration in a flat plate boundary layer diffusion flame (평판 경계층 확산화염에서의 국부적 가속현상에 관한 실험적 연구)

  • 심성훈;하지수;신현동
    • Transactions of the Korean Society of Mechanical Engineers
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    • v.12 no.4
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    • pp.847-864
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    • 1988
  • The main purpose of this study is to investigate the local velocity acceleration in a boundary layer diffusion flame over a flat plate. In order to know the effect of separation on the local velocity acceleration, two typical cases, flows with and without separation, are considered. For these cases, flow visualization using paraffine smoke tracers has been made. Mean velocity and r.m.s. value of fluctuating velocity are measured by using a laser Doppler velocimeter. In addition, measurements of time-mean concentration and time-mean temperature have been made. Time-mean density profiles have been obtained from the data of concentration and temperature. The obtained results are summarized as follows : (1) In the case without separation, the local velocity acceleration is clearly observed near the visible flame zone for all flow conditions. On the while, in the case with serration, the local velocity acceleration is observed only at low free stream velocity and high fuel injection velocity. As increasing the free stream velocity or decreasing the fuel injection velocity, it is not distinctly observed in the mean velocity profile. (2) The r.m.s. value of fluctuating velocity is significantly decreased by combustion in the case with separation. But in the case without separation, the r.m.s. value is increased near the visible flame zone in comparison with cold flow. In both cases, the peak value of r.m.s. appeared just at the visible flame zone, where the mean velocity gradient is not too high.

Characterization and Distribution of Transferrin from the Last Larval Haemolymph of Papilio xuthus (호랑나비 유충 혈림프 Transferrin의 특성과 분포)

  • Shin, Myung-Ja;Kim, Kyoung-Keun;Kim, Jeong-Sook;Lim, Jae-Hwan;Seo, Eul-Won
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1046-1052
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    • 2007
  • Transferrin is a molecule carrying iron to store and maintain for iron homeostasis of living organisms. In this study, we have purified transferrin, as an iron-binding protein, from the last larval haemolymph of Papilio xuthus by KBr density gradient ultracentrifugation and gel filtration (superose 6 HR) using fast protein liquid chromatography (FPLC) and transferrin containing iron was identified by Ferene S staining. The purified haemolymph transferrin was shown to have molecular mass of 78 and 80 kDa and amino acid composition of transferrin was rich in aspartic acid, valine, leucine and glutamic acid. With immuno-diffusion assay, we confirmed the existence of the transferrin in the haemo-lymph and fat body by detection of visible and clear positive reaction. From the quantitative comparison by rocket immuno-electrophoresis process, the amount of transferrin were increased in the haemolymph of 3 days after pupation and the whole 5 days after pupation. Here, with biochemical and immunohistochemical analysis, we speculate the relationship of transferrin between the physical characteristics and distribution during metamorphosis of P. xuthus.

Prediction of Chemotherapeutic Response in Unresectable Non-small-cell Lung Cancer (NSCLC) Patients by 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) Assay

  • Chen, Juan;Cheng, Guo-Hua;Chen, Li-Pai;Pang, Ting-Yuan;Wang, Xiao-Le
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3057-3062
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    • 2013
  • Background: Selecting chemotherapy regimens guided by chemosensitivity tests can provide individualized therapies for cancer patients. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt (MTS) assay is one in vitro assay which has become widely used to evaluate the sensitivity to anticancer agents. The aim of this study was to evaluate the clinical applicability and accuracy of MTS assay for predicting chemotherapeutic response in unresectable NSCLC patients. Methods: Cancer cells were isolated from malignant pleural effusions of patients by density gradient centrifugation, and their sensitivity to eight chemotherapeutic agents was examined by MTS assay and compared with clinical response. Results: A total of 37 patients participated in this study, and MTS assay produced results successfully in 34 patients (91.9%). The sensitivity rates ranged from 8.8% to 88.2%. Twenty-four of 34 patients who received chemotherapy were evaluated for in vitro-in vivo response analysis. The correlation between in vitro chemosensitivity result and in vivo response was highly significant (P=0.003), and the total predictive accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for MTS assay were 87.5%, 94.1%, 71.4%, 88.9%, and 83.3%, respectively. The in vitro sensitivity for CDDP also showed a significant correlation with in vivo response (P=0.018, r=0.522). Conclusion: MTS assay is a preferable in vitro chemosensitivity assay that could be use to predict the response to chemotherapy and select the appropriate chemotherapy regimens for unresectable NSCLC patients, which could greatly improve therapeutic efficacy and reduce unnecessary adverse effects.

First-principles Study of Graphene/Hexagonal Boron Nitride Stacked Layer with Intercalated Atoms

  • Sung, Dongchul;Kim, Gunn;Hong, Suklyun
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.185.2-185.2
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    • 2014
  • We have studied the atomic and electronic structure of graphene nanoribbons (GNRs) on a hexagonal boron nitride (h-BN) sheet with intercalated atoms using first-principles calculations. The h-BN sheet is an insulator with the band gap about 6 eV and then it may a good candidate as a supporting dielectric substrate for graphene-based nanodevices. Especially, the h-BN sheet has the similar bond structure as graphene with a slightly longer lattice constant. For the computation, we use the Vienna ab initio simulation package (VASP). The generalized gradient approximation (GGA) in the form of the PBE-type parameterization is employed. The ions are described via the projector augmented wave potentials, and the cutoff energy for the plane-wave basis is set to 400 eV. To include weak van der Waals (vdW) interactions, we adopt the Grimme's DFT-D2 vdW correction based on a semi-empirical GGA-type theory. Our calculations reveal that the localized states appear at the zigzag edge of the GNR on the h-BN sheet due to the flat band of the zigzag edge at the Fermi level and the localized states rapidly decay into the bulk. The open-edged graphene with a large corrugation allows some space between graphene and h-BN sheet. Therefore, atoms or molecules can be intercalated between them. We have considered various types of atoms for intercalation. The atoms are initially placed at the edge of the GNR or inserted in between GNR and h-BN sheet to find the effect of intercalated atoms on the atomic and electronic structure of graphene. We find that the impurity atoms at the edge of GNR are more stable than in between GNR and h-BN sheet for all cases considered. The nickel atom has the lowest energy difference of ~0.2 eV, which means that it is relatively easy to intercalate the Ni atom in this structure. Finally, the magnetic properties of intercalated atoms between GNR and h-BN sheet are investigated.

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THE INHIBITORY EFFECT OF STREPTOCOCCAL CELL WALL EXTRACTS ON STIMULATION OF LYMPHOCYTES (연쇄구균의 세포벽 단백질 추출물이 림프구 활성의 억제에 미치는 영향)

  • Sang, Hyoung-Sook;Jeong, Hee-Il;Oh, Se-Hong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.275-288
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    • 1995
  • The inhibitory effect of cell wall extracts of streptococci, have been investigated to know host-parasite relationship or pathogenesis of abscess formation. Streptococci isolated from the infected root canals were sonicated to get cell wall extracts which have been known as one of the factors of pyogenesis. Lymphocytes separated by density gradient were stimulated with phytohemagglutinin and exposed to cell wall extracts of Streptococcus sanguis, S. mitis, S. uberis, S. mutans (ATCC 10449), and S. faecalis (ATCC 19433). [$^3H$]-thymidine uptake of lymphocytes was analyzed with scintillation counter and lactate dehyrogenase (LD) activity was measured with autochemistry analyzer. S. faeealis had the strongest inhibitory effect. beginning at $100\;{\mu}g/ml$ concentration of sonic extracts. S. sanguis and S. mitis had inhibitory effect at $300\;{\mu}g/ml$, while S. uberis and S. mutans showed no inhibitory, effect on DNA syntheis even at $300\;{\mu}g/ml$. Each streptococci showed different inhibitory effect on the DNA synthesis of lymphocytes, which finding indicated wide spectrum of susceptibility of lymphocytes according to streptococcus spp. There were no significant difference of LD activities between control and each streptococcal extracts. Streptococcal sonic extracts did not affect the morphological findings or number of colonies activated lymphocytes. These finding suggested the inhibitory effect of sonic extract of streptococci to lymphocytes could be detected by DNA synthesis inhibition, not by cellular membrane damage.

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The Origin of Organic Matters Utilized by Soft Bottom Macrozoobenthos in Tongyeong (안정동위원소 분석을 이용한 통영 연질조하대 대형저서동물의 유기물 기원 추적)

  • Yun Sung-Gyu;Yoon Byoung-Sun;Paik Sang-Gyu;Kang Chang-Keun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.spc1
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    • pp.189-197
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    • 2006
  • To elucidate the organic matter sources in soft-bottom macrozoobenthos at Tongyeong, Korea, macrozoobenthos and their potential food sources were sampled in May 2004, and their C and N stable isotope compositions were analyzed. In total we collected 128 macrozoobenthic species, which ranged from 465 to 3,775 individuals/$m^2$(average 2,108 ind.$m^2$) in density and from 47.8 to 539.9 gwwt/$m^2$(average 366.0 gwwt/$m^2$) in biomass. Cluster and multi-dimensional scaling analyses indicated that the macrozoobenthic community was divided into two distinct groups. Coastal inner stations, where commercial fish cages were established, were dominated by Amphioplus ancistrotus, Scoletoma longifoila and Tharyx sp., whereas open sea stations were dominated by Chaetozone spinosa, Scoletoma longifolia and Capitella capitata. ${\delta}^{13}C$ values of sedimentary organic matter showed a distinct gradient in the range of -18.4 to $-15.2\%_{\circ}$, with a declining trend from the coastal inner stations to open sea stations. This probably reflects the settling rate of organic wastes such as feces and pellets from fish cages near the coastal inner stations. The macrozoobenthos showed a broad ${\delta}^{13}C$ range from -19.5 to $-8.9\%_{\circ}$ at the coastal inner stations, and a relatively narrow range from -21.3 to $-12.9\%_{\circ}$ at the open sea stations. ${\delta}^{13}C$ values of macrozoobenthos paralleled those of sedimentary organic matter, Our isotope results suggested that macrozoobenthos near the coastal inner stations used organic wastes derived from fish cages along with phytoplankton, whereas the macrozoobenthos near the open sea stations used organic wasters derived mainly from phytoplankton.

Studies on the Migratory Ability of Primordial Germ Cells from Embryonic Gonads at Different Developmental Stages in Quail (메추리의 발달 중 배자 생식선에서 분리한 원시생식세포의 이동능에 대한 연구)

  • D. K. Kim;G. H. Song;J. N. Kim;D. K. Jeong;K. D. Kim
    • Korean Journal of Poultry Science
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    • v.28 no.1
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    • pp.69-76
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    • 2001
  • Avian primordial germ cells (PGCs) originate from the epiblast and appear in the germinal crescent. These PGCs enter the developing blood vessels during stage 10∼12 (H&H), circulate in the blood stream, migrate into the developing gonadal anlage and differentiate into germ cells. However, it is not clear until when the migratory ability of PGC is maintained. This study was conducted to examine whether migratory ability is present in PGCs from the gonad at later embryonic developmental stages. In the present study, gonads were dissected from 5-, 6- and 10-day old quail embryos and treated with trypsin-EDTA. Gonadal PGCs (gPGCs) were purified by Ficoll-density-gradient-centrifugation and labeled with PKH26 fluorescent dye. The PKH26-labeled gPGCs were microinjected into the blood vessel of the recipient quail embryo. Manipulated recipients were incubated for 3 days, embedded in paraffin and sdctioned. The foreign gPGCs were detected by fluorescent and confocal laser microscopy. As a result, quail gPGCs, from 10, 6 and 5 day old embryos could migrate through the recipient blood stream at early stage and settle in the gonads. Thus, results suggest that gPGCs from upto 10-day old embryos keep properties seen in circulating PGC. Therefore, the PGCs of 10-day old embryonic gonads can be used for the tools of genetic manipulation.

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Thermal Conductivity of Saturated Unfrozen Kaolinite during Consolidation (포화된 카올리나이트의 압밀에 따른 열전도계수에 관한 연구)

  • Kim, Hak-Seung;Lee, Jang-Guen;Kang, Jae-Mo;Kim, Young-Seok;Hong, Seung-Seo
    • The Journal of Engineering Geology
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    • v.21 no.2
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    • pp.157-162
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    • 2011
  • Experimental tests have been performed to measure the thermal conductivity of unsaturated soils, and computational models have been widely used to predict thermal conductivity. However, measured values of the thermal conductivity of unsaturated soils cannot be compared with predicted values because of the gradient in moisture content within unsaturated soils. In this study, experimental consolidation tests on saturated unfrozen kaolinite were performed to investigate the effect of dry density and moisture content on thermal conductivity. The results were used to evaluate the validity of a model employed to calculate thermal conductivity.

Purification and Characterization of Ice Nucleating Proteins from Ice Nucleation-Active Bacteria (빙핵활성 세균으로부터 빙핵활성 단백질의 정제 및 특성)

  • Kim, Ki-Chung;Lee, Ung;Song, Dong-Up;Cho, Baik-Ho
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.99-108
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    • 1996
  • 3종의 빙핵세균 Peudomonas syringae 8401, Pseudomonas fuorescens 8701, Erwinia herbicola 8701의 세포 외막으로부터 아무런 변성제도 사용치 않고 sucrose density gradient centrifugation, Sephacryl gel filtration chromatography, DEAE-cellulose ion exchange chromatography, non-denaturing buffer를 이용한 PAGE, electroelution, SDS-PAGE를 통해 빙핵활성 단백질을 고도로 정제할 수 있었다. P. suringae와 P. fluorescens에서는 각각 3종류(155 kD, 75 kD, 50 kD)의 빙핵활성 단백질이, E. herbicola에서는 155 kD를 제외한 2종류(75 kD, 50 kD)의 빙핵활성 단백질은 이 연구를 통해 처음 밝혀진 것으로 , 지금까지 보고된 빙핵활성 단백질(150 KD 이상)보다는 훨씬 작은 것이다. 이는 빙핵활성을 나타내는 단백질의 기본단위는 이 실험의 결과만에 의하면 최대 50 kD임을 시사한다. 이들 단백질은 그 유래된 세균의 종류나 또는 단백질 분자량의 크기에 관계없이 모두 -5.5~7.5$^{\circ}C$에서 물을 동결시키는 높은 빙핵활성을 갖고 있었다. 이는 지금까지 보고된 어느 정제단백질의 빙핵활성보다 높은 것이다. 정제된 단백질의 빙핵활성은 trypsin 처리에 의해 상실되었고, pH6~8범위에서는 안정하였으며, pH5이하, pH9이상에서는 활성을 상실하였다. 보존온도에 대한 영향은 3$0^{\circ}C$이상이 되면 점차 활성이 감소하는 경향을 보이다 37$^{\circ}C$이상에서는 활성이 완전히 상실되었다. 금속이온으로서 Hg\ulcorner이온과 SDS에 의해 활성이 상실되었으나 phosphatidylinositol의 첨가에 의해서는 활성이 약간 증가(-1$^{\circ}C$)하였다.

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