• Title/Summary/Keyword: Binding media

Search Result 113, Processing Time 0.023 seconds

Effect of Electrolytes on the Saturable Binding of Morphine in Rat Brain Tissue (백서(白鼠) 뇌조직(腦組織)에서 Morphine의 Saturable Binding에 미치는 전해질(電解質)의 영향(影響))

  • Ko, Bok-Hyun;Chae, Soo-Wan;Cho, Kyu-Park
    • The Korean Journal of Pharmacology
    • /
    • v.18 no.2
    • /
    • pp.33-44
    • /
    • 1982
  • The binding in vitro of an opiate agonist, $(^3H)-morphine$, was studied using rat brain slices which were incubated in the modified Krebs-Henseleit bicarbonate buffer solution containing various concentrations of electrolytes with or without morphine, naloxone or morphine+naloxone at $4^{\circ}C$ for 24 hours. The binding of $(^3H)-morphine$ may be seperated into two component; one a saturable binding and the other nonsaturable. The saturable binding may be calculated from the differences in binding observed in the absence and presence of high concentration of morphine. The maximal saturable binding and $K_D$ value in the naive preparations were $0.32{\pm}0.02\;pmole/mg$ protein and $0.75{\pm}0.07\;nM$ respectively. The saturable binding of $(^3H)-morphine$ was significantly increased by low temperature-treatment, while $K_D$ value was not changed. Morphine in the incubation media significantly increased the saturable binding of $(^3H)-morphine$ and $K_D$ value. Naloxone also increased the maximal saturable binding of $(^3H)-morphine$ and $K_D$ value of the drug. Decrease of $K^+\;and\;Mg^{++}$, and addition of $Mn^{++}$ in the incubation media significantly increased the saturable binding of $(^3H)-morphine$, but decrease of $Na^+$and increase of $Ca^{++}$ in the incubation media did not influence the binding. The increment of the saturable binding of $(^3H)-morphine$ by nonlabeled morphine in the incubation media was notaffected by decrease of $Na^+,\;K^+\;or\;Mg^{++}$, or addition of $Mn^{++}$ into the incubation media, but was inhibited by increase of $Ca^{++}$ in the incubation media, while the increment of the saturable binding of $(^3H)-morphine$ was net observed by decrease of $Na^+,\;K^+\;or\;Mg^{++}$, or increase of $Ca^{++}$ in the incubation media. The above results indicate that change of opiate binding sites in quality, i.e. affinity, and quantity, i.e. number of binding sites, may occur by low temperature-treatment in the absence and presence of morphine or naloxone and that electrolytes play role of the changes of opiate binding sites.

  • PDF

Evaluation of normal morphology, DNA fragmentation, and hyaluronic acid binding ability of human spermatozoa after using four different commercial media for density gradient centrifugation

  • Lee, Dayong;Jee, Byung Chul
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.46 no.1
    • /
    • pp.8-13
    • /
    • 2019
  • Objective: Density gradient centrifugation (DGC) is frequently used to isolate high-motility fractions of spermatozoa. We compared the efficacy of four DGC media in terms of the percentage of morphologically normal spermatozoa, DNA fragmentation level, and hyaluronic acid (HA) binding ability. Methods: Thirty men with a total motile spermatozoa count > 80 million participated. Semen samples were divided into four aliquots, which were processed using PureSperm, PureCeption, Sidney, and SpermGrad media, respectively. The DNA fragmentation level was measured using the Halosperm assay kit and HA binding ability was measured using the HBA assay kit. Results: The mean percentage of morphologically normal spermatozoa was significantly enhanced after DGC using all four media (10.3%, 9.9%, 9.8%, and 10.7%, respectively; p< 0.05 for each when compared with 6.9% in raw semen). The DNA fragmentation level was significantly reduced after DGC using PureSperm, PureCeption, and SpermGrad media (6.0%, 6.5%, and 4.9%, respectively; p< 0.05 for each when compared with 11.2% in raw semen), but not after DGC using Sidney media (8.5%, p> 0.05). HA binding ability did not change after DGC using any of the four media. Conclusion: The four media were equally effective for obtaining a sperm fraction with highly motile, morphologically normal sperm. PureSperm, PureCeption, and SpermGrad media were equally effective for acquiring a sperm fraction with less DNA fragmentation.

Staphylococcal methicillin resistance expression under various growth conditions

  • Lee, Yoo-Nik;Ryoung, Poo-Ha;Lee, Young-Ik
    • Journal of Microbiology
    • /
    • v.35 no.2
    • /
    • pp.103-108
    • /
    • 1997
  • To improve the detection of methicillin resistant staphylococci, lowered incubation temperature (30.deg.) and inclusion of sodium chloride in media have been empirically recommended. However, in this study, we found that sodium chloride in Peptone-Yeast Extract-K$\_$2/HPO$\_$4/ (PYK) medium decreased methicillin minimum inhibitory concentrations. Divalent cations were shown to restore the expression of staphylococcal methicillin resistance. However, when it was determined by efficiency of plating, sodium chloride increased methicillin resistance expression on agar medium in which higher divalent cations were contained in the agar medium. The decrease of minimum inhibitory concentrations at 30.deg.C by sodium chloride occurred in Brain Heart Infusion but did not occur in other media investigated. Interestingly, both PYK and Brain Heart Infusion media had peptone, which contain cholic acids having detergent activities. Inclusion of sodium chloride in PYK caused a higher rate of autolysis. Penicillin binding protein 2a that has a low affinity to beta-lactam antibiotics, was highly inducible in methicillin resistant Staphylococcus epidermidis strains. In this study, we found that autolysins that are activated by the sodium chloride decreased the minimum inhibitory concentration at 30.deg.C, and peptidoglycan is weakened due to the presence of methicillin. Peptone in the media may aggravate the fragile cells. However, stabilization due to the presence of divalent cations and production of penicilin binding protein 2a increase the survival of staphylococci.

  • PDF

Estrogen Inhibits Bcl-2 Expression and Stimulates Apoptosis Mediated by 2,3,7,8-Tetrachlrodibenzo-p-dioxirn

  • Hwang, Sohyun;Such, Jaehong;Byun, Boo-Hyeong;Joe, Cheol O.
    • Toxicological Research
    • /
    • v.19 no.4
    • /
    • pp.325-330
    • /
    • 2003
  • The effects of estrogen on apoptosis induced by 2,3,7,8-tetrachlorodibenzo-p-doxin (TCDD) were examined in cultured MCF-7 cells. TCDD stimulated apoptosis and inhibited the expression of bcl-2 gene in MCF-7 cells grown in the media supplemented with 10% fetal bovine serum. However, TCDD failed to induce apoptosis if cells were grown in the media deprived of all estrogen-like compounds. Removal of estrogen-like compounds from the growth media also led to the activation of bcl-2 gene expression in cells treated with TCDD. Combined treatment of estrogen with TCDD abrogated the binding of Aryl hydrocarbon Receptor (AhR)-TCDD complex to Dioxin response element (DRE) of bcl-2 gene leading to the inhibition of bcl-2 gene expression as well as stimulation of apoptosis. The present study suggests that the binding of estrogen receptor (ER)-estrogen complex to the estrogen responsive element (E) interferes with the binding of AhR- TCDD complex to the DRE and inhibits the bcl-2 expression.

Hormonal Regulation of Insulin-Like Growth Factor Binding Protein Secretion by a Bovine Mammary Epithelial Cell Line

  • Kim, W.Y.;Chow, J.C.;Hanigan, M.D.;Calvert, C.C.;Ha, J.K.;Baldwin, R.L.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.10 no.2
    • /
    • pp.233-239
    • /
    • 1997
  • A mammary epithelial cell line (MAC-T) established as a model for lactation was utilized to identify and characterize effects of various hormones upon insulin-like growth factor binding protein secretion. Ligand and immunoblot analyses of conditioned media indicated that insulin-like growth factor binding protein-2 was secreted by MAC-T cells. Insulin-like growth factor-I stimulated insulin-like growth factor binding protein-2 secretion in a dose-dependent manner, but prolactin and bovine somatotropin did not alter insulin-like growth factor binding protein-2 secretion. Insulin increased and cortisol decreased insulin-like growth factor binding protein-2 secretion. Effects of insulin-like growth factor-I on insulin-like growth factor binding protein-2 secretion support previous studies using primary cultures of bovine mammary cells and bovine fibroblasts. Effects of cortisol and insulin on insulin-like growth factor binding protein-2 secretion may be explained by changes in protein synthesis. In addition, supraphysiological doses of insulin can cross-react with the insulin-like growth factor-I receptor and stimulate insulin-like growth factor binding protein-2 secretion. MAC-T cells provide a model system to study mechanisms that regulate local insulin-like growth factor-I bioactivity.

Characterization of Soluble Type Human Folate Receptor on Folate Binding and Transport (Soluble type의 human folate receptor의 folate 결합과 수송에 관한 특성)

  • Kim, Won-Sin
    • Toxicological Research
    • /
    • v.14 no.3
    • /
    • pp.371-377
    • /
    • 1998
  • We have cloned a soluble type human folate receptor(hFR type${\gamma}$) from human thymus cDNA library using the PCR amplification technique. To examine whether hFR type${\gamma}$ has a folate transport activity, CHO cells were transfected with the pcDNAhFR${\gamma}$ expression plasmid, and the stable cell line CHO/hFR${\gamma}$ expressing a high level of the hFR type${\gamma}$ was identified by northern and western blot analysis. The CHO/hFR${\gamma}$ cells produced a [$H^3$]folic acid binding protein in the culture medium. However, we couldn't detect any cell surface [$H^3$] folic acid binding and transport activities. The growth of the CHO/hFR${\gamma}$ cells was more rapidly inhibited than the wild type CHO cells in the low concentration folic acid media. These observations indicate that although soluble type human folate receptor can bind [$H^3$]folate, it does not involve in folate transport.

  • PDF

Versatilities of Calix[4]pyrrole Based Anion Receptors

  • Lee, Chang-Hee
    • Bulletin of the Korean Chemical Society
    • /
    • v.32 no.3
    • /
    • pp.768-778
    • /
    • 2011
  • Calixpyrroles and related macrocycles are non-planer synthetic anion receptors that have attracted considerable attentions in recent years. Although the synthesis of calix[4]pyrrole (known as meso-octamethylporphyrinogen) was reported more than 100 years ago, the anion binding properties were first discovered in 1996. The simple calix[4]pyrroles can be synthesized in single step in high yield by condensation of pyrrole with acetone. The compounds showed preferential binding for halide anions including fluoride, phosphate, carboxylate, and chloride in organic media. Efforts to improve the anion affinity of calix[4]pyrrole and to enhance its selectivity have led to the synthesis of a variety of new calixpyrrole derivatives. Among the various modifications, introduction of straps on one side of the calix[4]pyrroles are the most effective. Incorporation of aromatic rings other than pyrroles also exhibited interesting binding behaviour. Introduction of signalling units as part of the strapping element enable to detect the anions on chromogenic or fluorogenic fashion. Finding of the anion transport properties across the membrane and cytotoxic effects of the calix[4]pyrroles open new window for calixpyrrole-related research. The polymer-incorporated systems have also been employed as anion complexants in solvent-solvent extraction. These old, yet easy-to-make macrocycles have well advanced more recently with the discovery of the ion-pair complexation properties. In this review, the synthetic developments and anion binding properties of calixpyrroles for the last decades will be discussed and will cover the advances in calixpyrrole chemistry.

Biochemical Application of IgG Fc-binding peptide: From Biochip to Targeted Nano Carrier

  • Chung, Sang Jeon
    • Proceedings of the Korean Vacuum Society Conference
    • /
    • 2013.02a
    • /
    • pp.84-84
    • /
    • 2013
  • FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetratingpeptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells.

  • PDF

Staphylococcus aureus Siderophore-Mediated Iron-Acquisition System Plays a Dominant and Essential Role in the Utilization of Transferrin-Bound Iron

  • Park Ra Young;Sun Hui Yu;Choi Mi Hwa;Bai Young Hoon;Shin Sung Heui
    • Journal of Microbiology
    • /
    • v.43 no.2
    • /
    • pp.183-190
    • /
    • 2005
  • Staphylococcus aureus is known to be capable of utilizing transferrin-bound iron, via both siderophore­and transferrin-binding protein (named IsdA)-mediated iron-acquisition systems. This study was designed in order to determine which iron-acquisition system plays the essential or dominant role with respect to the acquisition of iron from human transfenin, in the growth of S. aureus. Holotransferrin (HT) and partially iron-saturated transferrin (PT), but not apotransferrin (AT), were found to stimulate the growth of S. aureus. S. aureus consumed most of the transferrin-bound iron during the exponential growth phase. Extracellular proteases were not, however, involved in the liberation of iron from transferrin. Transferrin-binding to the washed whole cells via IsdA was not observed during the culture. The expression of IsdA was observed only in the deferrated media with AT, but not in the media supplemented with PT or HT. In contrast, siderophores were definitely produced in the deferrated media with PT and HT, as well as in the media supplemented with AT. The siderophores proved to have the ability to remove iron directly from transferrin, but the washed whole cells expressing IsdA did not. In the bioassay, the growth of S. aureus on transferrin-bound iron was stimulated by the siderophores alone. These results demonstrate that the siderophore-mediated iron-acquisition system plays a dominant and essential role in the uptake of iron from transferrin, whereas the IsdA-mediated iron-acquisition system may play only an ancillary role in the uptake of iron from transferrin.

Phase Transition Characteristics in $Ge_xSb_{100-x}$ Film for Optical Storage Media

  • Park Tae-jin;Kang Myung-jin;Choi Se-young
    • 정보저장시스템학회:학술대회논문집
    • /
    • 2005.10a
    • /
    • pp.124-127
    • /
    • 2005
  • Rewritable optical memory devices such as an CD-RW and DVD+RW are data storage media, which take advantage of the different optical properties in the amorphous and crystalline states of phase change materials. The switching property, structural transformation, transformation kinetics and chemical bindings of $Ge_xSb_{100-x}$($6{\le}x{\le}$34) were studied to investigate the feasibility of applying $Ge_xSb_{100-x}$ alloys in optical memory. The $Ge_xSb_{100-x}$ thin film was deposited by RF magnetron co-sputtering system and phase change characteristics were investigated by X-ray diffraction (XRD), static tester, inductively coupled plasma atomic emission spectrometer (ICP-AES) and atomic force microscopy (AEM). Optimum fiim composition of $Ge_xSb_{100-x}$ was studied and its minimum time fur laser induced crystallization and optical contrast fur phase transition was performed. These results might be correlated with the binding energies between Ge and Sb, and indicate that $Ge_xSb_{100-x}$ have an potential far optical memory applications.

  • PDF