Academic literature on the topic 'Non-specific binding'
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Journal articles on the topic "Non-specific binding"
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Flintoft, Louisa. "Surveying non-specific binding." Nature Reviews Genetics 14, no. 10 (September 18, 2013): 676. http://dx.doi.org/10.1038/nrg3586.
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Chen, Shi-Jie. "Site-Specific Binding of Non-Site-Specific Ions." Biophysical Journal 116, no. 12 (June 2019): 2237–39. http://dx.doi.org/10.1016/j.bpj.2019.04.038.
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Zenei, Taira, and Terada Hiroshi. "Specific and non-specific ligand binding to serum albumin." Biochemical Pharmacology 34, no. 11 (June 1985): 1999–2005. http://dx.doi.org/10.1016/0006-2952(85)90322-3.
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Shimizu, Seishi, Steven Abbott, Katarzyna Adamska, and Adam Voelkel. "Quantifying non-specific interactionsvialiquid chromatography." Analyst 144, no. 5 (2019): 1632–41. http://dx.doi.org/10.1039/c8an02244e.
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Mendel, C. M., and D. B. Mendel. "‘Non-specific’ binding. The problem, and a solution." Biochemical Journal 228, no. 1 (May 15, 1985): 269–72. http://dx.doi.org/10.1042/bj2280269.
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The concept of ‘non-specific’ binding, as it relates to studies of the binding of hormones to their receptors, is reviewed. It is concluded that the most widely used operational definition, namely binding that is not displaceable by an excess of unlabelled ligand, is often inaccurate, resulting either in overestimation of the number of high-affinity receptors and underestimation of the affinity of a given hormone for its receptor, or in a curvilinear Scatchard plot suggesting (artifactually) the presence of negative co-operativity or multiple classes of binding sites. The general use of an alternative approach to non-specific binding, in which the non-specific component is assessed from an analysis of total binding, is advocated. The superiority of this approach is illustrated with data on the binding of high-density lipoproteins to their receptors.
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Oda, Masayuki, Koji Furukawa, Kazuhiro Ogata, Akinori Sarai, and Haruki Nakamura. "Thermodynamics of specific and non-specific DNA binding by the c-myb DNA-binding domain." Journal of Molecular Biology 276, no. 3 (February 1998): 571–90. http://dx.doi.org/10.1006/jmbi.1997.1564.
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de Bruijn, Peter, Inge M. Ghobadi Moghaddam-Helmantel, Walter J. Loos, Ron H. J. Mathijssen, and Erik A. C. Wiemer. "The issue of non-specific binding of cabazitaxel." Journal of Chromatography B 932 (August 2013): 74–75. http://dx.doi.org/10.1016/j.jchromb.2013.06.016.
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Taylor, Thomas J., Gerald W. Parker, Abram B. Fajer, and Kay A. Mereish. "Non-specific binding of palytoxin to plastic surfaces." Toxicology Letters 57, no. 3 (August 1991): 291–96. http://dx.doi.org/10.1016/0378-4274(91)90203-i.
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Smith, Phil M., Indorica Sutradhar, Maxwell Telmer, Rishikesh Magar, Amir Barati Farimani, and B. Reeja-Jayan. "Isolating Specific vs. Non-Specific Binding Responses in Conducting Polymer Biosensors for Bio-Fingerprinting." Sensors 21, no. 19 (September 22, 2021): 6335. http://dx.doi.org/10.3390/s21196335.
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A longstanding challenge for accurate sensing of biomolecules such as proteins concerns specifically detecting a target analyte in a complex sample (e.g., food) without suffering from nonspecific binding or interactions from the target itself or other analytes present in the sample. Every sensor suffers from this fundamental drawback, which limits its sensitivity, specificity, and longevity. Existing efforts to improve signal-to-noise ratio involve introducing additional steps to reduce nonspecific binding, which increases the cost of the sensor. Conducting polymer-based chemiresistive biosensors can be mechanically flexible, are inexpensive, label-free, and capable of detecting specific biomolecules in complex samples without purification steps, making them very versatile. In this paper, a poly (3,4-ethylenedioxyphene) (PEDOT) and poly (3-thiopheneethanol) (3TE) interpenetrating network on polypropylene–cellulose fabric is used as a platform for a chemiresistive biosensor, and the specific and nonspecific binding events are studied using the Biotin/Avidin and Gliadin/G12-specific complementary binding pairs. We observed that specific binding between these pairs results in a negative ΔR with the addition of the analyte and this response increases with increasing analyte concentration. Nonspecific binding was found to have the opposite response, a positive ΔR upon the addition of analyte was seen in nonspecific binding cases. We further demonstrate the ability of the sensor to detect a targeted protein in a dual-protein analyte solution. The machine-learning classifier, random forest, predicted the presence of Biotin with 75% accuracy in dual-analyte solutions. This capability of distinguishing between specific and nonspecific binding can be a step towards solving the problem of false positives or false negatives to which all biosensors are susceptible.
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Fowers, K. D., and J. Kopeček. "Development of a fibrinolytic surface: specific and non-specific binding of plasminogen." Colloids and Surfaces B: Biointerfaces 9, no. 6 (September 1997): 315–30. http://dx.doi.org/10.1016/s0927-7765(97)00034-9.
Full textDissertations / Theses on the topic "Non-specific binding"
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Shedge, Hemangi Y. "Specific and non-specific binding of proteins and nucleic acids on chemically modified reticulated vitreous carbon electrodes." Connect to this title online, 2009. http://etd.lib.clemson.edu/documents/1246559605/.
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McLure, James Alexander, and james mclure@flinders edu au. "Physicochemical determinants of the non-specific binding of drugs to human liver microsomes." Flinders University. Medicine, 2008. http://catalogue.flinders.edu.au./local/adt/public/adt-SFU20081102.165952.
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Accurate determination of the in vitro kinetic parameters Km (Michaelis constant) and Ki (inhibition constant) is critical for the quantitative prediction of in vivo drug clearance and the magnitude of inhibitory drug interactions. A cause of inaccuracy in vitro arises from the assumption that all drug added to an incubation mixture is available for metabolism or inhibition. Many drugs bind non-specifically to the membrane of the in vitro enzyme source.
The aims of this thesis were to: 1) investigate the comparative importance of lipophilicity (as log P), and pKa as determinants of the non-specific binding of drugs to human liver microsomes; 2) develop and validate an ANS fluorescence technique for measuring the non-specific binding of drugs to human liver microsomes; 3) characterise the non-specific binding of a large dataset of physicochemically diverse drugs using the ANS fluorescence procedure; 4) evaluate relationships between selected physicochemical characteristics and the extent of non-specific binding of drugs to human liver microsomes and; 5) computationally model the non-specific binding of drugs to discriminate between high binding (fu(mic) less than 0.5) and low binding (fu(mic) greater than 0.5) drugs.
The comparative binding of the basic drugs atenolol (log P = 0.1; fu(mic) = 1.00), of propranolol (log P = 3.1; fu(mic) = 0.36 - 0.84), and imipramine (log P = 4.8; fu(mic) = 0.42 - 0.82) suggested that lipophilicity is a major determinant of non-specific binding. In contrast, the comparative binding of diazepam (pKa = 3.3; fu(mic) = 0.69 - 0.80), a neutral compound; and the bases propranolol (pKa = 9.5; fu(mic) = 0.36 - 0.84) and lignocaine (pKa = 9.5; fu(mic) = 0.98), indicated that pKa was not a determinant of the extent of non-specific binding. The non-binding of lignocaine, a relatively lipophilic base, was unexpected and confirmed by the non-binding of the structurally related compounds bupivacaine and ropivacaine. These results implicated physicochemical characteristics other than lipophilicity and charge as important for the non-specific binding of drugs to human liver microsomes.
An assay based on 1-anilinonaphthalene-8-sulfonate (ANS) fluorescence was developed using the seven drugs employed in the initial study. Non-specific binding data from equilibrium dialysis and the ANS fluorescence methods were compared and a linear correlation (r2 = 0.92, p less than 0.01) was observed at drug concentrations of 100 and 200 micromolar. The approach was further validated by characterising the microsomal binding of nine compounds (bupropion, chloroquine, chlorpromazine, diflunisal, flufenamic acid, meclofenamic acid, mianserine, triflupromazine, and verapamil) using both binding methods (i.e. equilibrium dialysis and ANS fluorescence). A significant logarithmic relationship (r2 greater than or equal to 0.90) was demonstrated between fu(mic) and the modulus of ANS fluorescence for all drugs and for basic drugs alone at concentrations of 100 and 200 micromolar, while the acidic/neutral drugs showed a significant linear relationship (r2 greater than or equal to 0.84) at these two concentrations (p less than 0.01). The non binding of bupropion provided further evidence that physicochemical properties other than log P and charge were important for non-specific binding of drugs to human liver microsomes.
The ANS fluorescence technique was then used to characterise the non-specific binding of 88 physicochemically diverse compounds. In general, acids and neutrals bound to a low extent (fu(mic) greater than 0.5) whereas bases bound the full fu(mic) range (0.0001 to 1). Statistically significant relationships were observed between the non-specific binding of bases and log P, the number of hydrogen bond donors and hydrogen bond acceptors per molecule, and molecular mass.
Preliminary in silico modeling of the dataset generated by the ANS fluorescence technique, using the program ROCS, provided discrimination of all but one (itraconazole) of the high binding bases. However, there were 14 false positives, resulting in low overall prediction accuracy.
Taken together, the studies conducted in this thesis provide important insights into the physicochemical factors that determine the non-specific binding of drugs to human liver microsomes.
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Slowski, Kathryn Johanna. "Identification of a novel microRNA involved in non-specific binding to a decoy transcript." Thesis, University of British Columbia, 2016. http://hdl.handle.net/2429/59879.
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MicroRNAs are known to be upregulated or downregulated in various types of cancer, leading to changes in the expression of genes involved in cellular proliferation, anti-apoptosis, migration, and invasion. To study the effects of microRNA loss or gain in different neoplasms, numerous models have been described to decrease or increase expression of microRNAs, but the off-target effects of different methods have not been well investigated. I investigated the possibility of off-target effects in a model of miR-143 knockdown in myeloid leukemia cell lines that implemented a microRNA sponge, or decoy, as a method to reduce microRNA expression. The high expression of a sponge with repetitive sequence elements and low expression of the intended microRNA for knockdown, miR-143, created conditions with increased potential for non-specific microRNAs to bind to the sponge. Therefore, I investigated the potential binding sites present in the sponge and whether any novel microRNAs could bind to these sites. I found a number of potential candidates and eliminated them based on their likelihood of regulating protein targets and their resemblance to a microRNA in structure, leaving one potential candidate. I found genomic evidence of the existence of this novel microRNA, evolutionary conservation of function, and performed assays that confirmed the biological activity. Next, the original sponge was redesigned to inhibit the binding of the potential non-specific microRNA; miR-X, or the miR-143 binding sites were mutated to inhibit the binding of miR-143 and capture miR-X instead. This demonstrated that binding of non-specific microRNA could be abrogated and differential protein abundance specific to the knockdown of each microRNA separately was verified. I conclude that non-specific binding to the sponge is a distinct possibility in experiments using this method of microRNA knockdown, which needs to be taken into account when designing sponges in the future. This work also demonstrates that there remain novel microRNAs awaiting discovery.Medicine, Faculty of
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Child, Chloe Rose. "Determing structure-activity relationships between novel PET radiotracers and their non-specific binding properties." Thesis, Imperial College London, 2012. http://hdl.handle.net/10044/1/9576.
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The non-invasive imaging modality positron emission tomography (PET) is used extensively in clinical settings and is increasingly being used by the pharmaceutical industry in drug development. Molecules of biological interest are labelled with positron emitting isotopes e.g. 11C, allowing their biodistribution and kinetics to be followed in vivo. A major factor in the failure of radioligands is the magnitude of unwanted background signal, non-specific binding (NSB) obscuring binding to the desired target. Assumptions have previously been made as to the physiochemical and pharmacological properties of radioligands that can affect NSB. However, little work has been carried out to quantify NSB with regard to determining structure-activity relationships (SARs) in order to optimise efficient radiotracer discovery. Non-specific binding is a poorly understood process but is believed to be related to the non-saturable binding of labelled molecules with tissue membranes. In this work the synthesis of novel radiolabelled molecular libraries has been conducted, their physicochemical properties determined and their non-specific binding measured in vitro using autoradiographical and cell based mass spectrometry assay methods. Structure-activity relationships have been formed between partition coefficient properties, acid dissociation constants, interaction energies and molecular weight in order to determine the effect each of these properties has on non-specific binding. Traditionally lipophilicity, log P, of a radioligand is the main predictor to its non-specific binding properties. However from this work it has been shown that a single physicochemical property cannot be relied on to predict the NSB of a radioligand but multiple properties must be considered.
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Westerhult, David. "Weak Affinity Chromatography : Evaluation of Different Silica Supports for Protein Immobilizationand Effect of Mobile Phases Regarding Retentionand Non-specific Binding." Thesis, Linnéuniversitetet, Institutionen för naturvetenskap, NV, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-17968.
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Fragment based lead discovery (FBLD), where libraries of small fragments are screened andlater on developed to lead compounds, is an alternative to the classical drug discovery methods such as high trough-put screening. Weak affinity chromatography (WAC) is a new promising approach to the screening process of FBLD. WAC is performed by injections of fragments onto a high performance liquid chromatography (HPLC) column in which a protein is immobilized to a silica support. The retention of the injected fragments is correlated to the binding affinity of the fragments towards the immobilized protein. Immobilization capacity of three different silica materials with varying pore size (Kromasil240 Å, Nucleosil 1000 Å and Kromasil 300 Å) was evaluated by immobilization of trypsin. Retention of benzamidines on the trypsin columns was evaluated with different mobile phases. Contribution of non-specific binding in the interaction between the 4-aminobenzamidine and thrombin was estimated by frontal chromatography on a capillary columnusing PBS and PBS/acetonitrile as mobile phases. This study showed that the Kromasil 300 Å had a superior immobilization capacity of trypsin compared to the Kromasil 240 Å andthe Nucleosil 1000 Å (100 mg compared to 87.4 mg and 15.1 mg trypsin/g silica, respectively). However, the Nucleosil 1000 Å might be a more suitable support for the immobilization of larger proteins. Adding 5 % methanol or acetonitrile to the mobile phase resulted in a significant (p < 0.05) decreased retention of benzamidine fragments on the trypsin 240 Å column. Non-specific binding between thrombin and 4-ABA was not statistically significantly altered when 5 % acetonitrile was added to the mobile phase.
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Fouquier, d'Herouel Aymeric. "Statistical models of TF/DNA interaction." Licentiate thesis, Stockholm : Numerisk analys och datalogi, Numerical Analysis and Computer Science, Kungliga Tekniska högskolan, 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4633.
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Cowsill, Benjamin James. "The physics of pregnancy tests : a biophysical study of interfacial protein adsorption." Thesis, University of Manchester, 2012. https://www.research.manchester.ac.uk/portal/en/theses/the-physics-of-pregnancy-tests-a-biophysical-study-of-interfacial-protein-adsorption(538b8e9c-9111-4eb9-ac7d-e5e75110e315).html.
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Pregnancy tests and related immunoassays are heavily dependent on specific and non-specific protein adsorption. These interfacial processes are affected by many factors that influence the in situ conformations of interfacially immobilised antibodies. This thesis examines a number of representative features with dual polarisation interferometry (DPI) and neutron reflection (NR), thus combining real-time dynamic monitoring with high interfacial structural resolution. Bovine serum albumin (BSA) was initially used as a model system to compare the surface coverage and thickness measurements of DPI and NR. The results show that DPI and NR provided similar surface coverage data but the measured thicknesses differed at BSA concentrations above 0.1 mg/ml. This discrepancy arose from the adoption of the uniform-layer model used by DPI for data analysis and the greater thickness sensitivity of NR. A model pregnancy immunoassay was built in steps on a silica surface so that the adsorption of each protein could be accurately monitored. Both DPI and NR provided evidence of BSA insertion into the gaps on the surface between the antibody molecules. This suggests that BSA adsorption is an excellent method to block the non-specific adsorption of target antigens to the immunoassay test surface. A magnetic tweezer system was designed and built in order to measure the specific antibody/antigen binding force. The antibodies and antigens were used to immuno-link magnetic beads to the experimental surface before the immuno-links were broken by increasing the attractive force between the magnetic tweezers and beads. The force per antibody/antigen immuno-link was estimated to lie between the values of 13.6 pN and 43.8 pN.Immuno-link detachment as a function of time was investigated. It was found that the immuno-link comprised both a strong and a weak interaction. The dissociation constant of the strong antibody/antigen interaction was found to equal 3E-4 /s and had an interaction length of 0.06 nm. The low population of beads bound by the second, weaker interaction meant that it was not possible to obtain accurate values of the dissociation constant and bond length of the second weaker interaction.
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Richardson, Mandek. "Theoretical and Experimental Investigations to Improve the Performance of Surface Acoustic Wave (SAW) Biosensors." Scholar Commons, 2014. https://scholarcommons.usf.edu/etd/5566.
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The objective of this dissertation is to improve the performance of surface acoustic wave (SAW) biosensors for use in point-of-care-testing (POCT) applications. SAW biosensors have the ability to perform fast, accurate detection of an analyte in real time without the use of labels. However, the technology suffers from the inability to differentiate between specific and non-specific binding. Due to this limitation, direct testing of bodily fluids using SAW sensors to accurately determine an analyte's concentration is difficult. In addition, these sensors are challenged by the need to detect small concentrations of a biomarker that are typically required to give a clinical diagnosis. Sensitivity, selectivity and reliability are three critical aspects for any sensing platform. To improve sensitivity the delay path of a SAW sensor has been modified with microcavities filled with various materials. These filled cavities increased sensitivity by confining wave energy to the surface by way of constructive interference and waveguiding. Thus, the improved sensitivity will result in a lower limit of detection. In addition, insertion loss is decreased as a consequence of increased wave confinement to the surface. Sensor selectivity and reliability are adversely affected by non-specific binding of unwanted species present in a sample. To address this issue a multifunctional SAW sensor is presented. The sensor consists of two SAW delay lines oriented orthogonal to each on ST-quartz in order to generate two distinct wave modes. One wave mode is used for sensing while the other is used to remove loosely bound material. By using the same transduction mechanism for both removal and sensing, the sensor chip is simplified and complex electronics are avoided. The findings of this research involve the technological advances for SAW biosensors that make their use in POCT possible.
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Vasquez, Colins. "Allosteric effects of the GTP-specific binding site and a benzimidazole-derivative non-nucleoside inhibitor on the hepatitis C virus RNA-dependent RNA polymerase." Thesis, McGill University, 2009. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=40750.
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The hepatitis C virus (HCV) is a positive-strand RNA virus that belongs to the Flaviviridae family. HCV expresses the non-structural protein, NS5B, an RNA-dependent RNA polymerase required for genome replication. Here, we describe the allosteric properties of both the GTP-specific binding site (G-site) of the HCV polymerase and VIR-1327, a benzimidazole-derivative non-nucleoside inhibitor (NNI). Using biochemical assays, we identified certain G-site residues that affect NS5B activity, including serine-29, proline-495 and valine-499. We also demonstrated that VIR-1327 is a potent inhibitor with a resistance profile consisting of P495A and V499A mutants, but is non-competitive with regards to GTP. Moreover, biophysical results suggest that the mechanism of action of VIR-1327 is to prevent the formation of a stable enzyme-template complex. Further analysis of the G-site and NNIs will not only provide a better understanding of HCV replication, but also validate different allosteric sites as virus-specific targets for the development of anti-HCV therapeutics.Le virus de l’hépatite C (VHC) est un virus d’ARN de polarité positive qui appartient à la famille Flaviviridae. Le VHC exprime la protéine non-structurelle NS5B, une ARN polymérase dépendante de l’ARN indispensable à la réplication du génome virale. Nous décrivons ici les propriétés allostériques du site GTP-spécifique (site-G) du VHC polymérase, ainsi que les propriétés allostériques de VIR-1327, un inhibiteur non-nucléoside (INN) qui est dérivé du benzimidazole. En utilisant des tests biochimiques, nous avons identifié certains résidus du site-G qui influencent l’activité du NS5B, incluant sérine-29, proline-495 et valine-499. Nous avons également démontré que VIR-1327 est un puissant inhibiteur avec un profil de résistance comprenant les mutants P495A et V499A, mais qu’il est non-compétitif avec le GTP. De plus, des résultats biophysiques suggèrent que le méchanisme d’action de VIR-1327 est de prévenir la formation d’un complexe enzyme-matrice stable. D’autres analyses du site-G et des INN vont non seulement fournir une meilleure compréhension de la réplication du VHC, mais aussi valider différents sites allostériques comme étant des cibles virus-spécifiques pour le développement de thérapeutiques contre le VHC.
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Engqvist, Martin. "A generic capture assay for immunogenicity, using Biacore." Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-198767.
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The purpose of this investigation was to create and optimise a capture assay for the detectionof anti-drug antibodies (ADA) in human plasma, using Biacore. We also dealt with the nonspecificplasma binding to mouse-derived anti-biotin which may occur in the capture assay.By paying attention to these things we aimed at reaching as high sensitivity as possible for theADA detection. The capture assay also benefited and gained flexibility from using the same regenerationsolution irrespective of drug and from having a composition that minimises the risk ofdamaging drug epitopes.
Books on the topic "Non-specific binding"
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Jörres, Achim, Dietrich Hasper, and Michael Oppert. Non-dialytic management of the patient with acute kidney injury. Edited by Norbert Lameire. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199592548.003.0228.
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The main focus in the non-dialytic management of patients with acute kidney injury (AKI) is the prevention and treatment of complications.Nutritional support is an important aspect as many patients tend to be hypercatabolic, thus requiring adequate caloric intake, yet without administration of excessive fluid volumes. Inadequate nutrition in AKI may lead to enhanced production of urea nitrogen and azotaemia. However, hyperglycaemia is a frequent complication in these patients, often requiring continuous insulin therapy to achieve the recommended blood glucose target range of 110–150 mg/dL (6.11–8.33 mmol/L).Patients with AKI are prone to infections which are a common cause of death in this population. Careful search for and intensive treatment of infections is therefore of utmost importance, and antimicrobial chemotherapy must be initiated as early as possible, especially in patients with sepsis and AKI.Drug dosing in patients with AKI is complex and difficult. Residual kidney function can be highly variable and drug disposition may be altered due to changes in distribution volume, protein binding, and metabolism. Moreover, many drugs can be removed by renal replacement therapy (RRT). Therefore, adequate dosing must take into account the patient’s individual clinical characteristics, the specific pharmacokinetic/pharmacodynamic properties of the drug, and the mode and intensity of renal replacement therapy.
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Pauwelyn, Joost. Sources of International Trade Law. Edited by Samantha Besson and Jean d’Aspremont. Oxford University Press, 2018. http://dx.doi.org/10.1093/law/9780198745365.003.0048.
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This chapter argues that the World Trade Organization (WTO) approach to sources of law is legal-positivist, non-teleological, and focused predominantly on the text of WTO covered agreements as explicitly agreed to by WTO members. This approach places heavy reliance on a de facto rule of precedent and an increasing role for non-binding instruments, with little or no reference to academic writings and a limited role for non-WTO rules of international law other than mainly procedural rules of general international law. Moreover, the WTO’s sources doctrine remains relatively traditional or mainstream. It is difficult to speak of a WTO- or trade-specific ‘deviation’ from the general rule of recognition regarding the establishment of sources. At the same time, the WTO experience does have specific features, with a more prominent role for some sources over others and some pushing of the boundaries when it comes to certain less traditional sources of international law.
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Roy, Goode, Kronke Herbert, and McKendrick Ewan, eds. Part III Harmonization of General Contract Law, 16 Restatements of Contract Law. Oxford University Press, 2015. http://dx.doi.org/10.1093/law/9780198735441.003.0017.
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This chapter examines non-binding restatements of contract law, in particular the UNIDROIT Principles of International Commercial Contracts and the Principles of European Contract Law. It considers the nature, purposes, scope, sphere of application, and substantive content of these Principles (including freedom of contract, pacta sunt servanda, good faith, interpretation, adequate assurance of performance, specific performance, and other remedies and hardship and change of circumstances). The chapter considers the extent to which these Principles can be used in litigation and in arbitration and their relative advantages and disadvantages.
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Berger, Tobias. Between the State and the Shalish. Oxford University Press, 2017. http://dx.doi.org/10.1093/oso/9780198807865.003.0004.
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This chapter reconstructs the background knowledge that underpins all processes of conflict resolution in rural Bangladesh. It starts with the in-depth analysis of an attempted murder case and shows how state and non-state courts are intricately intertwined in what is conceptualized as the logic of non-enforcement. According to this logic, people frequently file cases with the state court, but not to get binding and enforceable verdicts. Instead, they do so to alter the dynamics of non-state courts. Although non-state justice institutions respond to representations of the state (for example in the form of official documents), they nonetheless do not conform to state law. Instead, their decisions are informed by specific normative convictions about the importance of social harmony and religious prescripts. Importantly, this logic of non-enforcement also applies to the state’s Muslim family laws, which are frequently not enforced if they threaten the patriarchal order of things in rural Bangladesh.
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Papadopoulos, Yannis. Multilevel Governance and Depoliticization. Oxford University Press, 2017. http://dx.doi.org/10.1093/oso/9780198748977.003.0007.
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‘Multilevel’ governance (MLG) refers to the fact that, in contemporary established democracies, collectively binding decisions are frequently formulated or implemented in a cooperative manner by networks composed of public actors attached to different jurisdictional levels (from the local to the supranational) and of non-public actors such as experts, interest representatives, and members of cause groups. This chapter develops the expectation that the occurrence and magnitude of depoliticization in MLG depend on a number of its defining traits, and that the presence and intensity of these traits depend in turn on the specific empirical configuration and actor constellation of governance arrangements. The chapter first lays out the relationships that may exist between different facets of depoliticization in MLG, and then explores how MLG is depoliticized when technocratic rule, deficits of representation, lack of political control, and lack of public debate tend to prevail.
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James, Harrison. 1 Introduction. Oxford University Press, 2017. http://dx.doi.org/10.1093/law/9780198707325.003.0001.
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Chapter 1 provides an introduction to both the anthropogenic threats facing the marine environment at the beginning of the twenty-first century and the role that international law plays in regulating humankind’s impacts on the oceans. It argues that the protection of the marine environment is a common concern of humankind, requiring the cooperation of all States in adopting appropriate international rules and standards to address the main threats to the marine environment and collective efforts to ensure the enforcement of those norms. The chapter then explains the key sources of international law that are relevant to the regulation of marine activities, highlighting the central role played by treaties, related non-binding instruments, judicial decisions, and general principles of international law. This introduction to the sources of international law provides a basic background to the more detailed analysis of specific treaties and related instruments in the remainder of the book.
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Das, Onita, and Aneaka Kellayv. Private Security Companies and Other Private Security Service Providers (PSCs) and Environmental Protection in Jus Post Bellum. Oxford University Press, 2017. http://dx.doi.org/10.1093/oso/9780198784630.003.0014.
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A challenge to environmental protection and the jus post bellum framework is the rise in Private Security Companies and other Private Security Service Providers (PSCs). The marked increase in the outsourcing of vast amounts of operational and logistical work to PSCs have caused key issues around PSC oversight, regulation, and concern around civilian protection linked to environmental issues to arise. Using the Iraq (2003–11) and Afghanistan (2001–14) conflicts as examples, this chapter explores the growth of PSCs, their environmental performance, and reviews the adequacy of legal and policy frameworks that regulate PSCs to ensure the provision of adequate environmental protection as part of jus post bellum in order to contribute to sustainable peace. Areas of law explored include international humanitarian law, international human rights law, binding legislation and soft law specific to PSCs, contract litigation, corporate liability, state and non-state actor obligations in respect to PSCs, and shared responsibility.
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Kohn, Robert. Human Rights and the Elderly. Oxford University Press, 2017. http://dx.doi.org/10.1093/med/9780199374656.003.0042.
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Attention to the human rights of the elderly has gained increased international attention. This chapter provides an overview of some international agencies’ efforts to address human rights of elders, as well as related efforts in different regions of the world. The UN does not have a specific treaty or convention regarding human rights of the elderly; only in 1995 was comprehensive legal analysis of the rights of the elderly addressed. The 2002 Madrid International Action Plan on Aging was a non-legally binding declaration by participating governments. Article 14 addresses equal access to healthcare and services, including physical and mental health services. The Commission on Human Rights 2011 Special Rapporteur emphasized human rights issues in primary healthcare and chronic illness; long-term care; palliative care; and informed consent as related to older persons. The Alzheimer’s Disease International Kyoto Declaration provides countries with a framework of action to address the needs and quality of life of those with dementia and their caregivers. Similarly, the United States has developed a national plan to address Alzheimer’s disease.
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Faarlund, Jan Terje. The Syntax of Mainland Scandinavian. Oxford University Press, 2019. http://dx.doi.org/10.1093/oso/9780198817918.001.0001.
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The term Mainland Scandinavian covers the North Germanic languages spoken in Denmark, Norway, Sweden, and parts of Finland. There is a continuum of mutually intelligible standard languages, regional varieties, and dialects stretching from southern Jutland to Eastern Finland. Linguistically, Danish, Norwegian, and Swedish are thus to be considered one language. Most syntactic patterns and features are shared among the national and regional varieties, but there are also interesting differences. This book presents the main syntactic structures of this language, with the focus on the standard languages, but some widespread or typologically interesting non-standard phenomena are included. This is mainly a descriptive work, with a minimum of technical formalities and theoretical discussion. The theoretical background and descriptive framework is generative grammar in its current version, known as ‘minimalism’. The minimalist architecture partly determines the ‘bottom-up’ organization of the book, with separate chapters or subchapters dealing with each of the phrase types, starting with the lexical phrases. After an introductory chapter, chapter 2 deals with the noun phrase and the determiner phrase. Chapters 3–5 deal with lexical phrase types with adjectives, prepositions. and verbs as their heads. Chapter 6 deals with the TP domain, and chapter 7 with the CP domain. The last three chapters deal with more specific topics, subordination, anaphor binding, and conjunction, and ellipsis.
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Gill, Steven J., and Michael H. Nathanson. Central nervous system pathologies and anaesthesia. Edited by Philip M. Hopkins. Oxford University Press, 2017. http://dx.doi.org/10.1093/med/9780199642045.003.0081.
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Anaesthesia induces changes in many organ systems within the body, though clearly none more so than the central nervous system. The physiology of the normal central nervous system is complex and the addition of chronic pathology and polypharmacy creates a significant challenge for the anaesthetist. This chapter demonstrates a common approach for the anaesthetist and specific considerations for a wide range of neurological conditions. Detailed preoperative assessment is essential to gain understanding of the current symptomatology and neurological deficit, including at times restrictions on movement and position. Some conditions may pose challenges relating to communication, capacity, and consent. As part of the consent process, patients may worry that an anaesthetic may aggravate or worsen their neurological disease. There is little evidence to support this understandable concern; however, the risks and benefits must be considered on an individual patient basis. The conduct of anaesthesia may involve a preference for general or regional anaesthesia and requires careful consideration of the pharmacological and physiological impact on the patient and their disease. Interactions between regular medications and anaesthetic drugs are common. Chronically denervated muscle may induce hyperkalaemia after administration of succinylcholine. Other patients may have an altered response to non-depolarizing agents, such as those suffering from myasthenia gravis. The most common neurological condition encountered is epilepsy. This requires consideration of the patient’s antiepileptic drugs, often relating to hepatic enzyme induction or less commonly inhibition and competition for protein binding, and the effect of the anaesthetic technique and drugs on the patient’s seizure risk. Postoperative care may need to take place in a high dependency unit, especially in those with limited preoperative reserve or markers of frailty, and where the gastrointestinal tract has been compromised, alternative routes of drug delivery need to be considered. Overall, patients with chronic neurological conditions require careful assessment and preparation, a considered technique with attention to detail, and often higher levels of care during their immediate postoperative period.
Book chapters on the topic "Non-specific binding"
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Meijer, Ellen A., Sacco C. de Vries, Peter Sterk, Dorus W. J. Gadella, Karel W. A. Wirtz, and Theo Hendriks. "Characterization of the non-specific lipid transfer protein EP2 from carrot (Daucus carota L.)." In Cellular Fatty Acid-Binding Proteins II, 159–66. Boston, MA: Springer US, 1993. http://dx.doi.org/10.1007/978-1-4615-3096-1_21.
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Wangkam, Thidarat, and Toemsak Srikhirin. "Design Sensor Chip for Repelling the Non-specific Binding for Its Application in Biomedical Sensor." In IFMBE Proceedings, 857–60. Cham: Springer International Publishing, 2014. http://dx.doi.org/10.1007/978-3-319-02913-9_221.
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Vorwerk, P., K. Mohnike, W. F. Blum, U. Kluba, V. Aumann, and U. Mittler. "Insulin-Like Growth Factors and Their Specific Binding Proteins in Leukemia (ALL) and Non-Hodgkin Lymphoma." In Acute Leukemias VI, 161–65. Berlin, Heidelberg: Springer Berlin Heidelberg, 1997. http://dx.doi.org/10.1007/978-3-642-60377-8_28.
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Korth, Ruth-Maria. "Non-Specific PAF Binding to Embryonal F9 Cells and Prostacyclin Synthesis in Human Umbilical Vein Endothelial Cells." In Advances in Experimental Medicine and Biology, 291–95. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4899-0179-8_46.
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Allfrey, Vincent G., Akira Inoue, Jonathan Karn, Edward M. Johnson, Robert A. Good, and John W. Hadden. "Sequence-Specific Binding of DNA by Non-Histone Proteins and Their Migration from Cytoplasm to Nucleus During Gene Activation." In Novartis Foundation Symposia, 199–228. Chichester, UK: John Wiley & Sons, Ltd, 2008. http://dx.doi.org/10.1002/9780470720103.ch12.
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Levitan, Irena, Sang Joon Ahn, Ibra Fancher, and Avia Rosenhouse-Dantsker. "Physiological Roles and Cholesterol Sensitivity of Endothelial Inwardly-Rectifying K+ Channels: Specific Cholesterol-Protein Interactions Through Non Annular Binding Sites." In Vascular Ion Channels in Physiology and Disease, 327–47. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/978-3-319-29635-7_15.
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Prakash, Om, and Feroz Khan. "CoSSDb: A Database of Co-crystallized Ligand Sub-structures for Anticancer Lead Designing & Optimization." In Proceedings of the Conference BioSangam 2022: Emerging Trends in Biotechnology (BIOSANGAM 2022), 133–41. Dordrecht: Atlantis Press International BV, 2022. http://dx.doi.org/10.2991/978-94-6463-020-6_14.
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AbstractThe Discovery of the novel optimized structures of small molecules for selective targeting is one of the challenging tasks in drug designing. Bioisosteres are the key components of the lead compound, which provide hidden power to the compound scaffold for selective targeting. We are presenting a database, named CoSSDb which stands for Co-crystallized Sub-Structure Database. The CoSSDb contains ligand sub-structures as possible bioisosteres. extracted from PDB files, available in Protein Data Bank. Sub-structures were extracted through an algorithm, which utilizes the location of atoms in the 3D domain of the complex ligand & protein. It processes the relative positioning of atoms for demarcation of the influential part of the ligand, which interacts with macromolecule and provides potency to that ligand for binding with a specific binding pocket of the protein. The algorithm was used to extract sub-structures from the ligands co-crystallized with proteins involved in cancer. About 7721 x-ray crystallography PDB files were processed, and 654 non-redundant substructures were identified. These sub-structures will be useful during designing & optimization of novel ligands for selective targets. The database is freely accessible at ‘https://opticket49.wixsite.com/substructdb’.
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Arnoldi, Michele, Giulia Zarantonello, Stefano Espinoza, Stefano Gustincich, Francesca Di Leva, and Marta Biagioli. "Design and Delivery of SINEUP: A New Modular Tool to Increase Protein Translation." In Methods in Molecular Biology, 63–87. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2010-6_4.
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AbstractSINEUP is a new class of long non-coding RNAs (lncRNAs) which contain an inverted Short Interspersed Nuclear Element (SINE) B2 element (invSINEB2) necessary to specifically upregulate target gene translation. Originally identified in the mouseAS-Uchl1 (antisense Ubiquitin carboxyl-terminal esterase L1) locus, natural SINEUP molecules are oriented head to head to their sense protein coding, target gene (Uchl1, in this example). Peculiarly, SINEUP is able to augment, in a specific and controlled way, the expression of the target protein, with no alteration of target mRNA levels. SINEUP is characterized by a modular structure with the Binding Domain (BD) providing specificity to the target transcript and an effector domain (ED)—containing the invSINEB2 element—able to promote the loading to the heavy polysomes of the target mRNA. Since the understanding of its modular structure in the endogenous AS-Uchl1 ncRNA, synthetic SINEUP molecules have been developed by creating a specific BD for the gene of interest and placing it upstream the invSINEB2 ED. Synthetic SINEUP is thus a novel molecular tool that potentially may be used for any industrial or biomedical application to enhance protein production, also as possible therapeutic strategy in haploinsufficiency-driven disorders.Here, we describe a detailed protocol to (1) design a specific BD directed to a gene of interest and (2) assemble and clone it with the ED to obtain a functional SINEUP molecule. Then, we provide guidelines to efficiently deliver SINEUP into mammalian cells and evaluate its ability to effectively upregulate target protein translation.
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Magraw, Kendra. "Trends and ISDS Backlash Related to Non-Disputing Treaty Party Submissions." In Public Actors in International Investment Law, 79–96. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-58916-5_5.
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AbstractSome international investment agreements (IIAs) allow states that are parties to a treaty, but are not party to a specific dispute under that treaty, to intervene on a limited basis in order to make submissions on matters of treaty interpretation. Such mechanisms have proved to be highly valued by treaty parties, as evident by the many recently-concluded IIAs containing increasingly sophisticated non-disputing treaty party (NDTP) provisions. This chapter: (1) provides the background on NDTPs mechanisms, with a focus on the North American Free Trade Agreement (NAFTA) (the first-known IIA to contain such a provision); (2) examines the possible connection between tribunals failing to give due regard to treaty parties’ interpretive positions (again focusing on NAFTA) and the current backlash against investor-state dispute settlement (ISDS); and (3) analyses trends in recently-concluded IIAs. It is argued that the apparent lack of deference given by tribunals to NDTP submissions may be contributing to the current backlash against ISDS, based on two discernible trends: (1) an increase in the number of IIAs containing NDTPs provisions; and (2) provisions that now state that not only are treaty interpretations made by treaty parties binding on tribunals (such provisions also have their genesis in NAFTA), but that, in addition, tribunals’ decisions must be consistent with such agreed interpretations (the latter an innovation of a NAFTA party in 2003). Such trends are also visible at the institutional and multilateral levels, such as the revision of the Arbitration Rules of the International Centre for Settlement of Investment Disputes (ICSID) and the Mauritius Convention on Transparency in ISDS of the United Nations Commission on International Trade Law (UNCITRAL), and show no sign of slowing down.
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Danisi, Carmelo, Moira Dustin, Nuno Ferreira, and Nina Held. "Why Sexual Orientation and Gender Identity Asylum?" In IMISCOE Research Series, 3–21. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-69441-8_1.
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AbstractForced migration – no matter how we wish to define it – has been high in the political agendas and debates across the world for several decades. Forced migrants become claimants of international protection, or ‘asylum claimants’, and then find themselves trapped in convoluted, constraining and highly politicised systems. Often accused of being ‘bogus’ asylum claimants, they are also regularly accused of abusing the hospitality of the host country, violating countries’ borders and territorial sovereignty, and simply seeking economic benefits (Ford 2009; UNHCR 2007). Conversely, asylum legal instruments have been repeatedly criticised for inadequately addressing the rights and needs of asylum claimants, therefore preventing those with legitimate claims from being granted protection. These debates have more recently been rehashed in the context of the negotiations behind the Global Compact for Safe Orderly and Regular Migration, a non-legally binding agreement negotiated under the aegis of the United Nations (UN) and endorsed by the UN General Assembly. In this atmosphere of permanent politicised and humanitarian ‘crisis’ (McAdam 2014), a group warranting specific attention is constituted by those asylum claimants presenting a claim based on their sexual orientation or gender identity (SOGI).
Conference papers on the topic "Non-specific binding"
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King, Benjamin C., Michael Clark, Thomas Burkhead, Palaniappan Sethu, Shesh Rai, Goetz Kloecker, and Balaji Panchapakesan. "Electrical detection of specific versus non-specific binding events in breast cancer cells." In SPIE NanoScience + Engineering, edited by Hooman Mohseni, Massoud H. Agahi, and Manijeh Razeghi. SPIE, 2012. http://dx.doi.org/10.1117/12.929875.
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Tsukiiwa, Mirano, Hironaga Noguchi, and Yuhei Hayamizu. "Self-assembled peptides for suppression of non-specific binding of biomolecules on graphene." In 2020 International Conference on Solid State Devices and Materials. The Japan Society of Applied Physics, 2020. http://dx.doi.org/10.7567/ssdm.2020.g-8-02.
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Zainudin, Nor Syuhada, Nor Azura Malini Ahmad Hambali, Mohamad Halim Abd Wahid, Vithyacharan Retnasamy, and Mukhzeer Mohamad Shahimin. "The study of non-fouling and non-specific cellular binding on functionalized surface for mammalian cell identification and manipulation." In ADVANCED MATERIALS ENGINEERING AND TECHNOLOGY V: International Conference on Advanced Material Engineering and Technology 2016. Author(s), 2017. http://dx.doi.org/10.1063/1.4981840.
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Wang, Qian, Min Zhang, Ling Wang, and Yueqing Gu. "Effective reduction of non-specific binding by bovine serum albumin modified quantum dot surface for cell targeted imaging." In Twelfth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2014), edited by Qingming Luo, Lihong V. Wang, and Valery V. Tuchin. SPIE, 2014. http://dx.doi.org/10.1117/12.2068693.
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Zheng, Xiangjun, Siu Lun Cheung, Lian Wang, Joyce A. Schroeder, Ronald L. Heimark, James C. Baygents, Roberto Guzman, and Yitshak Zohar. "Specific Binding of Cancer Cells Using a Microchamber Array Functionalized With Antibodies." In ASME 2009 International Mechanical Engineering Congress and Exposition. ASMEDC, 2009. http://dx.doi.org/10.1115/imece2009-13217.
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Specific binding of target suspended metastatic cancer cells to an antibody-functionalized surface utilizing a microfluidic device has experimentally been investigated under various conditions. The microfluidic devices, fabricated in silicon using DRIE process, consisted of a 5×5 micochamber array; each 1mm×1mm in area, and 50μm in depth. The oxide surface of the microchammber array was functionalized with various antibodies immobilized on a protein G layer. The microfluidic device design allows accurate counting of cells loaded into each microchamber and, thus, enabling a reliable counting of cells captured from homogeneous suspensions. The effects of cell suspension concentration, incubation times and ambient temperature on cell capture efficiency have been examined. Furthermore, to evaluate the specificity of the cell-surface interaction, several cell cancer types expressing different membrane receptors have been incubated on surfaces functionalized with various counter receptors. Specific binding of up to 100% of the suspended cells is observed when using surfaces functionalized with counter receptors matching the cell receptors; otherwise, non-specific binding of less than 15% of suspended cells is obtained if the functionalized counter receptors do not match the cell receptors.
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Wirth, Rachel, Sangeetha Rajagopalan, Erin Willert, William Null, Jennifer Erdman, Jen-Sing Liu, and Jack P. Higgins. "Abstract 5477: Engineered toxin body demonstrating CD20-specific binding and cell kill in B-cell non-Hodgkin's lymphoma cells." In Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC. American Association for Cancer Research, 2013. http://dx.doi.org/10.1158/1538-7445.am2013-5477.
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Bowry, S. K., and M. Tepel. "NON-REVERSIBLE BINDING OF 5'-p-FLUOROSULFONYLBENZOYL ADENOSINE TO WASHED INTACT PLATELETS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643506.
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ADP is an important in vivo mediator of platelet aggregation. The identification and estimation of a finite number of receptor sites has been made difficult by the reversible nature of the action of ADP on platelets and because ADP can be degraded by enzymes on the platelet surface. Analogues of ADP have therefore been commonly used to study the ADP receptor. We investigated the validity of using 5'-p-fluorosulfonylbenzoyl adenosine (FSBA), an affinity analogue of ADP that inhibits ADP-induced aggregation, for studying the ADP receptor on intact washed platelets. Binding of labelled FSBA, like for ADP, was saturable. Scatchard plot analyses of equilibrium binding data indicated, positive cooperativity; the apparent affinity (2.4 × 10−6) correlated well with the amount of ADP required to cause aggregation and with the value obtained from aggregation studies. However, the platelet bound FSBA was not displaceable with excess unlabelled FSBA indicating non-reversibility of ligand binding. As the minimal criteria for receptor identification (specificity, saturability and reversibility) are not fulfilled, the usage of FSBA to study the ADP receptor must be treated with caution. As FSBA is able to bind covalently to at least four polypeptides in isolated platelet membranes, a finite number of receptors cannot be postulated using FSBA. Furthermore, positive cooperativity, as indicated by the Scatchard plots, may be explained in terms of specific non-receptor binding. As the ADP receptor cannot be identified with any degree of certainty, the possibility therfore exists that the mechanism by which ADP initiates aggregation by rearrangement of platelet membrane proteins without binding to a single protein receptor.
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Bowry, S. X. "ENHANCED ASSOCIATION OF FIBRINOGEN WITH ITS PLATELET RECEPTOR DUE TO SODIUM CITRATE INDICATION FOR ONE FIBRINOGEN RECEPTOR ONLY." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643701.
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Washed platelet suspensions are almost always prepared from blood anticoagulated with sodium citrate. As citrate has been implicated to affect platelet function and as estimates of the number of fibrinogen (Fbg) bindingsites on platelets range from 4,700 -82,500, we examined the involvement of citrate on the platelet-fibrinogen interaction. The binding of 125-1-fibrinogen to washed platelets from citrated blood (PCB) and thosefrom gel-filtered non-anticoagulated blood (PNB) from the same donor showed linear Scatchard plots for PNB and curvilinear plots for PCB. Assuming the presence of two classes of bindingsites, a high_.affinity site (5,540±760 molecules/ platelets; Kd, 1.29 x 10-7 M) and low affinity site (32,070 α 6,520 molecules/platelet; Kd 1.02 x 10-6 M) were determined for PCB. However, PNB indicated a single class of binding sites with 16,480α 2,800 Fbg molecules/platelet.When blood from one donor was collected into 10 mM and 20 mM citrate, increased binding of Fbg was observed onplatelets exposed to 20 mM citrate. The effects of citrate appear to be on theplatelet Fbg receptor since non-specific binding was not affected by the citrate. As no 14-C-citric acid binding to platelets was observed, citrate may affect the receptors without binding to the platelets. The dependence of binding on the pHof thecitrate suggests ionic interactions between platelets and citrate. Different amounts of Fbg were bound when four different preparations of sodium citrate varying in concentration and pH were used. Ourdata suggest that acombination of thedirect effects of citrate on the Fbg bindingof platelets, together with the variable concentrations and pH of the different citrate preparations routinelyused toanticoagulate blood, may explain why some investigators obtain upwardly concave Scatchard plotswhile others obtain linear plots. Ourresultsindicate that the major reasonfor the disparities in the Fbg binding datais due to the effects of citrate on platelets.
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Lian, E. C. Y., and F. A. Siddigui. "BINDING OF 37-DKa PLATELET AGGLUTINATING PROTEIN TO HUMAN PLATELETS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643976.
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We have previously reported the purification of a 37-KDa platelet agglutinating protein (PAP p37) from the plasma of a patient with thrombotic thrombocytopenic purpura. using 125I-labeled p37, the properties of its binding to platelets were studied. The binding of p37 to washed human platelets from 4 normal subjects and two TTP patients after recovery was specific, concentration dependent and saturable. The Scatchard analysis revealed that the binding sites for p37 was about 100,000 per platelet with a dissociation constant of 48 × 10−9 M. The binding of p37 to erythrocytes was very little and non-specific. Stimulation of platelets by thrombin or ADP did not have any effect on the binding of p37 to platelets. The monoclonal antibodies to platelet GP lb (6D1) and GP Ilb-llla (10E5)(A gift of Dr. Barry coller) did not inhibit the binding of p37 to platelets. Fibrinogen (1 mg/ml) and FVIII/vWF (250 ug/ml) reduced the binding slightly. The polyclonal antibodies to p37 as well as concanavalin-A inhibited the binding of p37 to platelets through their direct interaction with p37. Other lectins such as phytohemagglutinin, potato lectin and helix pomatia lectin did not have any effect. At 40 mM, sialic acid, α-D-(+)-glucose, D-(+)-mannose and D-fructose caused 91%,44%,79%, and 63% inhibition of p37 binding respectively. D-(+)-galactose did not interfere with the binding. It is concluded that p37 binds to platelets on the sites other than GP lb and Gp IIb-IIIa and its binding to platelets is inhibited by certain sugars.
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Cho, Hongkwan, Abdul Sheikh, and Daria A. Narmoneva. "Non-Specific Endothelial Cell Interactions With the Substrate Result in Cell Activation and Angiogenesis In Vitro." In ASME 2010 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2010. http://dx.doi.org/10.1115/sbc2010-19094.
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Vascularization is critical for success of tissue engineering applications. Previous studies by us and others have shown that self-assembling peptide nanoscaffold RAD16-II promotes de novo capillary formation (angiogenesis) in vitro and neovascularization in vivo, and is a promising material for tissue engineering applications [1, 2]. However, the molecular mechanisms for cell interactions with this material are not known. Angiogenesis is mediated via interactions between integrins, which are expressed on the surface of activated endothelial cells (ECs), and extracellular matrix proteins. Among several integrins, αvβ3 is the most abundant and influential receptor regulating angiogenesis [3]. The αvβ3 integrin binds to its ligands via Arg-Gly-Asp (RGD) biding motif. However, there are no RGD motifs on RAD 16-II peptide. Instead, it contains three RAD motifs. Studies have shown that non-specific binding of αvβ3 with RAD can be retained through R and D sides [4]. The objective of this study, therefore, is to elucidate the underlying molecular mechanisms of RAD16-II nanoscaffold interactions with microvascular endothelial cells. We hypothesize that non-specific interactions between RAD16-II peptide nanoscaffold and αvβ3 integrin result in phosphorylations of β3 cytoplasmic domain, which then activate downstream angiogenic signaling pathways and promote angiogenesis.
Reports on the topic "Non-specific binding"
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Gurevitz, Michael, Michael E. Adams, Boaz Shaanan, Oren Froy, Dalia Gordon, Daewoo Lee, and Yong Zhao. Interacting Domains of Anti-Insect Scorpion Toxins and their Sodium Channel Binding Sites: Structure, Cooperative Interactions with Agrochemicals, and Application. United States Department of Agriculture, December 2001. http://dx.doi.org/10.32747/2001.7585190.bard.
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Integrated pest management in modern crop protection may combine chemical and biological insecticides, particularly due to the risks to the environment and livestock arising from the massive use of non-selective chemicals. Thus, there is a need for safer alternatives, which target insects more specifically. Scorpions produce anti-insect selective polypeptide toxins that are biodegradable and non-toxic to warm-blooded animals. Therefore, integration of these substances into insect pest control strategies is of major importance. Moreover, clarification of the molecular basis of this selectivity may provide valuable information pertinent to their receptor sites and to the future design of peptidomimetic anti-insect specific substances. These toxins may also be important for reducing the current overuse of chemical insecticides if they produce a synergistic effect with conventional pesticides. Based on these considerations, our major objectives were: 1) To elucidate the three-dimensional structure and toxic-site of scorpion excitatory, "depressant, and anti-insect alpha toxins. 2) To obtain an initial view to the sodium channel recognition sites of the above toxins by generating peptide decoys through a phage display system. 3) To investigate the synergism between toxins and chemical insecticides. Our approach was to develop a suitable expression system for toxin production in a recombinant form and for elucidation of toxin bioactive sites via mutagenesis. In parallel, the mode of action and synergistic effects of scorpion insecticidal toxins with pyrethroids were studied at the sodium channel level using electrophysiological methods. Objective 1 was achieved for the alpha toxin, LqhaIT Zilberberg et al., 1996, 1997; Tugarinov et al., 1997; Froy et al., 2002), and the excitatory toxin, Bj-xtrIT (Oren et al., 1998; Froy et al., 1999; unpublished data). The bioactive surface of the depressant toxin, LqhIT2, has been clarified and a crystal of the toxin is now being analyzed (unpublished). Objective 2 was not successful thus far as no phages that recognize the toxins were obtained. We therefore initiated recently an alternative approach, which is introduction of mutations into recombinant channels and creation of channel chimeras. Objective 3 was undertaken at Riverside and the results demonstrated synergism between LqhaIT or AaIT and pyrethroids (Lee et al., 2002). Furthermore, negative cross-resistance between pyrethroids and scorpion toxins (LqhaIT and AaIT) was demonstrated at the molecular level. Although our study did not yield a product, it paves the way for future design of selective pesticides by capitalizing on the natural competence of scorpion toxins to distinguish between sodium channels of insects and vertebrates. We also show that future application of anti-insect toxins may enable to decrease the amounts of chemical pesticides due to their synergism.
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Rafaeli, Ada, and Russell Jurenka. Molecular Characterization of PBAN G-protein Coupled Receptors in Moth Pest Species: Design of Antagonists. United States Department of Agriculture, December 2012. http://dx.doi.org/10.32747/2012.7593390.bard.
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The proposed research was directed at determining the activation/binding domains and gene regulation of the PBAN-R’s thereby providing information for the design and screening of potential PBAN-R-blockers and to indicate possible ways of preventing the process from proceeding to its completion. Our specific aims included: (1) The identification of the PBAN-R binding domain by a combination of: (a) in silico modeling studies for identifying specific amino-acid side chains that are likely to be involved in binding PBAN with the receptor and; (b) bioassays to verify the modeling studies using mutant receptors, cell lines and pheromone glands (at tissue and organism levels) against selected, designed compounds to confirm if compounds are agonists or antagonists. (2) The elucidation ofthemolecular regulationmechanisms of PBAN-R by:(a) age-dependence of gene expression; (b) the effect of hormones and; (c) PBAN-R characterization in male hair-pencil complexes. Background to the topic Insects have several closely related G protein-coupled receptors (GPCRs) belonging to the pyrokinin/PBAN family, one with the ligand pheromone biosynthesis activating neuropeptide or pyrokinin-2 and another with diapause hormone or pyrokinin-1 as a ligand. We were unable to identify the diapause hormone receptor from Helicoverpa zea despite considerable effort. A third, related receptor is activated by a product of the capa gene, periviscerokinins. The pyrokinin/PBAN family of GPCRs and their ligands has been identified in various insects, such as Drosophila, several moth species, mosquitoes, Triboliumcastaneum, Apis mellifera, Nasoniavitripennis, and Acyrthosiphon pisum. Physiological functions of pyrokinin peptides include muscle contraction, whereas PBAN regulates pheromone production in moths plus other functions indicating the pleiotropic nature of these ligands. Based on the alignment of annotated genomic sequences, the primary and secondary structures of the pyrokinin/PBAN family of receptors have similarity with the corresponding structures of the capa or periviscerokinin receptors of insects and the neuromedin U receptors found in vertebrates. Major conclusions, solutions, achievements Evolutionary trace analysisof receptor extracellular domains exhibited several class-specific amino acid residues, which could indicate putative domains for activation of these receptors by ligand recognition and binding. Through site-directed point mutations, the 3rd extracellular domain of PBAN-R was shown to be critical for ligand selection. We identified three receptors that belong to the PBAN family of GPCRs and a partial sequence for the periviscerokinin receptor from the European corn borer, Ostrinianubilalis. Functional expression studies confirmed that only the C-variant of the PBAN-R is active. We identified a non-peptide agonist that will activate the PBAN-receptor from H. zea. We determined that there is transcriptional control of the PBAN-R in two moth species during the development of the pupa to adult, and we demonstrated that this transcriptional regulation is independent of juvenile hormone biosynthesis. This transcriptional control also occurs in male hair-pencil gland complexes of both moth species indicating a regulatory role for PBAN in males. Ultimate confirmation for PBAN's function in the male tissue was revealed through knockdown of the PBAN-R using RNAi-mediated gene-silencing. Implications, both scientific and agricultural The identification of a non-peptide agonist can be exploited in the future for the design of additional compounds that will activate the receptor and to elucidate the binding properties of this receptor. The increase in expression levels of the PBAN-R transcript was delineated to occur at a critical period of 5 hours post-eclosion and its regulation can now be studied. The mysterious role of PBAN in the males was elucidated by using a combination of physiological, biochemical and molecular genetics techniques.
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Altstein, Miriam, and Ronald Nachman. Rationally designed insect neuropeptide agonists and antagonists: application for the characterization of the pyrokinin/Pban mechanisms of action in insects. United States Department of Agriculture, October 2006. http://dx.doi.org/10.32747/2006.7587235.bard.
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The general objective of this BARD project focused on rationally designed insect neuropeptide (NP) agonists and antagonists, their application for the characterization of the mechanisms of action of the pyrokinin/PBAN (PK-PBAN) family and the development of biostable, bioavailable versions that can provide the basis for development of novel, environmentally-friendly pest insect control agents. The specific objectives of the study, as originally proposed, were to: (i) Test stimulatory potencies of rationally designed backbone cyclic (BBC) peptides on pheromonotropic, melanotropic, myotropic and pupariation activities; (ii) Test the inhibitory potencies of the BBC compounds on the above activities evoked either by synthetic peptides (PBAN, LPK, myotropin and pheromonotropin) or by the natural endogenous mechanism; (iii) Determine the bioavailability of the most potent BBC compounds that will be found in (ii); (iv) Design, synthesize and examine novel PK/PBAN analogs with enhanced bioavailability and receptor binding; (v) Design and synthesize ‘magic bullet’ analogs and examine their ability to selectively kill cells expressing the PK/PBAN receptor. To achieve these goals the agonistic and antagonistic activities/properties of rationally designed linear and BBC neuropeptide (NP) were thoroughly studied and the information obtained was further used for the design and synthesis of improved compounds toward the design of an insecticide prototype. The study revealed important information on the structure activity relationship (SAR) of agonistic/antagonistic peptides, including definitive identification of the orientation of the Pro residue as trans for agonist activity in 4 PK/PBANbioassays (pheromonotropic, pupariation, melanotropic, & hindgut contractile) and a PK-related CAP₂b bioassay (diuretic); indications that led to the identification of a novel scaffold to develop biostbiostable, bioavailable peptidomimetic PK/PBANagonists/antagonists. The work led to the development of an arsenal of PK/PBAN antagonists with a variety of selectivity profiles; whether between different PKbioassays, or within the same bioassay between different natural elicitors. Examples include selective and non-selective BBC and novel amphiphilic PK pheromonotropic and melanotropic antagonists some of which are capable of penetrating the moth cuticle in efficacious quantities. One of the latter analog group demonstrated unprecedented versatility in its ability to antagonize a broad spectrum of pheromonotropic elicitors. A novel, transPro mimetic motif was proposed & used to develop a strong, selective PK agonist of the melanotropic bioassay in moths. The first antagonist (pure) of PK-related CAP₂b diuresis in flies was developed using a cisPro mimetic motif; an indication that while a transPro orientation is associated with receptor agonism, a cisPro orientation is linked with an antagonist interaction. A novel, biostablePK analog, incorporating β-amino acids at key peptidase-susceptible sites, exhibited in vivo pheromonotropic activity that by far exceeded that of PBAN when applied topically. Direct analysis of neural tissue by state-of-the-art MALDI-TOF/TOF mass spectrometry was used to identify specific PK/PK-related peptides native to eight arthropod pest species [house (M. domestica), stable (S. calcitrans), horn (H. irritans) & flesh (N. bullata) flies; Southern cattle fever tick (B. microplus), European tick (I. ricinus), yellow fever mosquito (A. aegypti), & Southern Green Stink Bug (N. viridula)]; including the unprecedented identification of mass-identical Leu/Ile residues and the first identification of NPs from a tick or the CNS of Hemiptera. Evidence was obtained for the selection of Neb-PK-2 as the primary pupariation factor of the flesh fly (N. bullata) among native PK/PK-related candidates. The peptidomic techniques were also used to map the location of PK/PK-related NP in the nervous system of the model fly D. melanogaster. Knowledge of specific PK sequences can aid in the future design of species specific (or non-specific) NP agonists/antagonists. In addition, the study led to the first cloning of a PK/PBAN receptor from insect larvae (S. littoralis), providing the basis for SAR analysis for the future design of 2ⁿᵈgeneration selective and/or nonselective agonists/antagonists. Development of a microplate ligand binding assay using the PK/PBAN pheromone gland receptor was also carried out. The assay will enable screening, including high throughput, of various libraries (chemical, molecular & natural product) for the discovery of receptor specific agonists/antagonists. In summary, the body of work achieves several key milestones and brings us significantly closer to the development of novel, environmentally friendly pest insect management agents based on insect PK/PBANNPs capable of disrupting critical NP-regulated functions.
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Kelly, Luke. Characteristics of Global Health Diplomacy. Institute of Development Studies (IDS), June 2021. http://dx.doi.org/10.19088/k4d.2021.09.
Full textAbstract:
This rapid review focuses on Global Health Diplomacy and defines it as a method of interaction between the different stakeholders of the public health sector in a bid to promote representation, cooperation, promotion of the right to health and improvement of health systems for vulnerable populations on a global scale. It is the link between health and international relations. GHD has various actors including states, intergovernmental organizations, private companies, public-private partnerships and non-governmental organizations. Foreign policies can be integrated into national health in various ways i.e., designing institutions to govern practices regarding health diplomacy (i.e., health and foreign affairs ministries), creating and promoting norms and ideas that support foreign policy integration and promoting policies that deal with specific issues affecting the different actors in the GHD arena to encourage states to integrate them into their national health strategies. GHD is classified into core diplomacy – where there are bilateral and multilateral negotiations which may lead to binding agreements, multistakeholder diplomacy – where there are multilateral and bilateral negotiations which do not lead to binding agreements and informal diplomacy – which are interactions between other actors in the public health sector i.e., NGOs and Intergovernmental Organizations. The US National Security Strategy of 2010 highlighted the matters to be considered while drafting a health strategy as: the prevalence of the disease, the potential of the state to treat the disease and the value of affected areas. The UK Government Strategy found the drivers of health strategies to be self-interest (protecting security and economic interests of the state), enhancing the UK’s reputation, and focusing on global health to help others. The report views health diplomacy as a field which requires expertise from different disciplines, especially in the field of foreign policy and public health. The lack of diplomatic expertise and health expertise have been cited as barriers to integrating health into foreign policies. States and other actors should collaborate to promote the right to health globally.
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Gurevitz, Michael, William A. Catterall, and Dalia Gordon. face of interaction of anti-insect selective toxins with receptor site-3 on voltage-gated sodium channels as a platform for design of novel selective insecticides. United States Department of Agriculture, December 2013. http://dx.doi.org/10.32747/2013.7699857.bard.
Full textAbstract:
Voltage-gated sodium channels (Navs) play a pivotal role in excitability and are a prime target of insecticides like pyrethroids. Yet, these insecticides are non-specific due to conservation of Navs in animals, raising risks to the environment and humans. Moreover, insecticide overuse leads to resistance buildup among insect pests, which increases misuse and risks. This sad reality demands novel, more selective, insect killers whose alternative use would avoid or reduce this pressure. As highly selective insect toxins exist in venomous animals, why not exploit this gift of nature and harness them in insect pest control? Many of these peptide toxins target Navs, and since their direct use via transformed crop plants or mediator microorganisms is problematic in public opinion, we focus on the elucidation of their receptor binding sites with the incentive of raising knowledge for design of toxin peptide mimetics. This approach is preferred nowadays by agro-industries in terms of future production expenses and public concern. However, characterization of a non-continuous epitope, that is the channel receptor binding site for such toxins, requires a suitable experimental system. We have established such a system within more than a decade and reached the stage where we employ a number of different insect-selective toxins for the identification of their receptor sites on Navs. Among these toxins we wish to focus on those that bind at receptor site-3 and inhibit Nav inactivation because: (1) We established efficient experimental systems for production and manipulation of site-3 toxins from scorpions and sea anemones. These peptides vary in size and structure but compete for site-3 on insect Navs. Moreover, these toxins exhibit synergism with pyrethroids and with other channel ligands; (2) We determined their bioactive surfaces towards insect and mammalian receptors (see list of publications); (3) We found that despite the similar mode of action on channel inactivation, the preference of the toxins for insect and mammalian channel subtypes varies greatly, which can direct us to structural features in the basis of selectivity; (4) We have identified by channel loop swapping and point mutagenesis extracellular segments of the Navinvolved with receptor site-3. On this basis and using channel scanning mutagenesis, neurotoxin binding, electrophysiological analyses, and structural data we offer: (i) To identify the residues that form receptor site-3 at insect and mammalian Navs; (ii) To identify by comparative analysis differences at site-3 that dictate selectivity toward various Navs; (iii) To exploit the known toxin structures and bioactive surfaces for modeling their docking at the insect and mammalian channel receptors. The results of this study will enable rational design of novel anti-insect peptide mimetics with minimized risks to human health and to the environment. We anticipate that the release of receptor site-3 molecular details would initiate a worldwide effort to design peptide mimetics for that site. This will establish new strategies in insect pest control using alternative insecticides and the combined use of compounds that interact allosterically leading to increased efficiency and reduced risks to humans or resistance buildup among insect pests.
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Diop, Ahmed. Country Diagnostic Study – Senegal. Islamic Development Bank Institute, October 2021. http://dx.doi.org/10.55780/rp21003.
Full textAbstract:
The Country Diagnostic Study (CDS) for Senegal uses the Hausmann-Rodrik-Velasco growth diagnostics model to identify the binding constraints being faced in its quest for higher and more sustained economic growth and make recommendations to relax these constraints. Hence, the findings of the CDS can help the Islamic Development Bank in identifying areas where it can have a greater impact and provide an evidence-basis to support the development of the Member Country Partnership Strategy. After decades of subdued and highly volatile economic growth due to heavy dependence on primary commodities and low productivity, Senegal experienced an unprecedented growth acceleration from 2014 to 2019. However, there appeared to be a weak correlation between economic growth and jobs creation. In addition, about 90 percent of non-agricultural employment is estimated to be informal. The national poverty rate decreased by 5 percentage points between 2011 and 2018. Nonetheless, the absolute number of poor people has increased. Furthermore, regional disparities are persistent. Despite the country’s solid performance in the field of governance, further simplification and transparency of business procedures and regulations will be critical in addressing the challenge of informality. Efforts to address informality in the economy should also target the issue of access to finance through the design of financing mechanisms based on specific needs assessment and risk management tools. Senegal will also need to create the conditions for higher competitiveness and follow upgrading trajectories in global and regional value chains. In this respect, both physical and digital connectivity will be essential.
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Grumet, Rebecca, and Benjamin Raccah. Identification of Potyviral Domains Controlling Systemic Infection, Host Range and Aphid Transmission. United States Department of Agriculture, July 2000. http://dx.doi.org/10.32747/2000.7695842.bard.
Full textAbstract:
Potyviruses form one of the largest and most economically important groups of plant viruses. Individual potyviruses and their isolates vary in symptom expression, host range, and ability to overcome host resistance genes. Understanding factors influencing these biological characteristics is of agricultural importance for epidemiology and deployment of resistance strategies. Cucurbit crops are subject to severe losses by several potyviruses including the highly aggressive and variable zucchini yellow mosaic virus (ZYMV). In this project we sought to investigate protein domains in ZYMV that influence systemic infection and host range. Particular emphasis was on coat protein (CP), because of known functions in both cell to cell and long distance movement, and helper component-protease (HC-Pro), which has been implicated to play a role in symptom development and long distance movement. These two genes are also essential for aphid mediated transmission, and domains that influence disease development may also influence transmissibility. The objectives of the approved BARD project were to test roles of specific domains in the CP and HC-Pro by making sequence alterations or switches between different isolates and viruses, and testing for infectivity, host range, and aphid transmissibility. These objectives were largely achieved as described below. Finally, we also initiated new research to identify host factors interacting with potyviral proteins and demonstrated interaction between the ZYMV RNA dependent RNA polymerase and host poly-(A)-binding protein (Wang et al., in press). The focus of the CP studies (MSU) was to investigate the role of the highly variable amino terminus (NT) in host range determination and systemic infection. Hybrid ZYMV infectious clones were produced by substituting the CP-NT of ZYMV with either the CP-NT from watermelon mosaic virus (overlapping, but broader host range) or tobacco etch virus (TEV) (non- overlapping host range) (Grumet et al., 2000; Ullah ct al., in prep). Although both hybrid viruses initially established systemic infection, indicating that even the non-cucurbit adapted TEV CP-NT could facilitate long distance transport in cucurbits, after approximately 4-6, the plants inoculated with the TEV-CPNT hybrid exhibited a distinct recovery of reduced symptoms, virus titer, and virus specific protection against secondary infection. These results suggest that the plant recognizes the presence of the TEV CP-NT, which has not been adapted to infection of cucurbits, and initiates defense responses. The CP-NT also appears to play a role in naturally occurring resistance conferred by the zym locus in the cucumber line 'Dina-1'. Patterns of virus accumulation indicated that expression of resistance is developmentally controlled and is due to a block in virus movement. Switches between the core and NT domains of ZYMV-NAA (does not cause veinal chlorosis on 'Dina-1'), and ZYMV-Ct (causes veinal chlorosis), indicated that the resistance response likely involves interaction with the CP-NT (Ullah and Grumet, submitted). At the Volcani Center the main thrust was to identify domains in the HC-Pro that affect symptom expression or aphid transmissibility. From the data reported in the first and second year report and in the attached publications (Peng et al. 1998; Kadouri et al. 1998; Raccah et al. 2000: it was shown that: 1. The mutation from PTK to PAK resulted in milder symptoms of the virus on squash, 2. Two mutations, PAK and ATK, resulted in total loss of helper activity, 3. It was established for the first time that the PTK domain is involved in binding of the HC-Pro to the potyvirus particle, and 4. Some of these experiments required greater amount of HC-Pro, therefore a simpler and more efficient purification method was developed based on Ni2+ resin.
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Barg, Rivka, Erich Grotewold, and Yechiam Salts. Regulation of Tomato Fruit Development by Interacting MYB Proteins. United States Department of Agriculture, January 2012. http://dx.doi.org/10.32747/2012.7592647.bard.
Full textAbstract:
Background to the topic: Early tomato fruit development is executed via extensive cell divisions followed by cell expansion concomitantly with endoreduplication. The signals involved in activating the different modes of growth during fruit development are still inadequately understood. Addressing this developmental process, we identified SlFSM1 as a gene expressed specifically during the cell-division dependent stages of fruit development. SlFSM1 is the founder of a class of small plant specific proteins containing a divergent SANT/MYB domain (Barg et al 2005). Before initiating this project, we found that low ectopic over-expression (OEX) of SlFSM1 leads to a significant decrease in the final size of the cells in mature leaves and fruits, and the outer pericarp is substantially narrower, suggesting a role in determining cell size and shape. We also found the interacting partners of the Arabidopsis homologs of FSM1 (two, belonging to the same family), and cloned their tomato single homolog, which we named SlFSB1 (Fruit SANT/MYB–Binding1). SlFSB1 is a novel plant specific single MYB-like protein, which function was unknown. The present project aimed at elucidating the function and mode of action of these two single MYB proteins in regulating tomato fruit development. The specific objectives were: 1. Functional analysis of SlFSM1 and its interacting protein SlFSB1 in relation to fruit development. 2. Identification of the SlFSM1 and/or SlFSB1 cellular targets. The plan of work included: 1) Detailed phenotypic, histological and cellular analyses of plants ectopically expressing FSM1, and plants either ectopically over-expressing or silenced for FSB1. 2) Extensive SELEX analysis, which did not reveal any specific DNA target of SlFSM1 binding, hence the originally offered ChIP analysis was omitted. 3) Genome-wide transcriptional impact of gain- and loss- of SlFSM1 and SlFSB1 function by Affymetrix microarray analyses. This part is still in progress and therefore results are not reported, 4) Search for additional candidate partners of SlFSB1 revealed SlMYBI to be an alternative partner of FSB1, and 5) Study of the physical basis of the interaction between SlFSM1 and SlFSB1 and between FSB1 and MYBI. Major conclusions, solutions, achievements: We established that FSM1 negatively affects cell expansion, particularly of those cells with the highest potential to expand, such as the ones residing inner to the vascular bundles in the fruit pericarp. On the other hand, FSB1 which is expressed throughout fruit development acts as a positive regulator of cell expansion. It was also established that besides interacting with FSM1, FSB1 interacts also with the transcription factor MYBI, and that the formation of the FSB1-MYBI complex is competed by FSM1, which recognizes in FSB1 the same region as MYBI does. Based on these findings a model was developed explaining the role of this novel network of the three different MYB containing proteins FSM1/FSB1/MYBI in the control of tomato cell expansion, particularly during fruit development. In short, during early stages of fruit development (Phase II), the formation of the FSM1-FSB1 complex serves to restrict the expansion of the cells with the greatest expansion potential, those non-dividing cells residing in the inner mesocarp layers of the pericarp. Alternatively, during growth phase III, after transcription of FSM1 sharply declines, FSB1, possibly through complexing with the transcription factor MYBI serves as a positive regulator of the differential cell expansion which drives fruit enlargement during this phase. Additionally, a novel mechanism was revealed by which competing MYB-MYB interactions could participate in the control of gene expression. Implications, both scientific and agricultural: The demonstrated role of the FSM1/FSB1/MYBI complex in controlling differential cell growth in the developing tomato fruit highlights potential exploitations of these genes for improving fruit quality characteristics. Modulation of expression of these genes or their paralogs in other organs could serve to modify leaf and canopy architecture in various crops.
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Morrison, Mark, Joshuah Miron, Edward A. Bayer, and Raphael Lamed. Molecular Analysis of Cellulosome Organization in Ruminococcus Albus and Fibrobacter Intestinalis for Optimization of Fiber Digestibility in Ruminants. United States Department of Agriculture, March 2004. http://dx.doi.org/10.32747/2004.7586475.bard.
Full textAbstract:
Improving plant cell wall (fiber) degradation remains one of the highest priority research goals for all ruminant enterprises dependent on forages, hay, silage, or other fibrous byproducts as energy sources, because it governs the provision of energy-yielding nutrients to the host animal. Although the predominant species of microbes responsible for ruminal fiber degradation are culturable, the enzymology and genetics underpinning the process are poorly defined. In that context, there were two broad objectives for this proposal. The first objective was to identify the key cellulosomal components in Ruminococcus albus and to characterize their structural features as well as regulation of their expression, in response to polysaccharides and (or) P AA/PPA. The second objective was to evaluate the similarities in the structure and architecture of cellulosomal components between R. albus and other ruminal and non-ruminal cellulolytic bacteria. The cooperation among the investigators resulted in the identification of two glycoside hydrolases rate-limiting to cellulose degradation by Ruminococcus albus (Cel48A and CeI9B) and our demonstration that these enzymes possess a novel modular architecture specific to this bacterium (Devillard et al. 2004). We have now shown that the novel X-domains in Cel48A and Cel9B represent a new type of carbohydrate binding module, and the enzymes are not part of a ceiluiosome-like complex (CBM37, Xu et al. 2004). Both Cel48A and Cel9B are conditionally expressed in response to P AA/PPA, explaining why cellulose degradation in this bacterium is affected by the availability of these compounds, but additional studies have shown for the first time that neither PAA nor PPA influence xylan degradation by R. albus (Reveneau et al. 2003). Additionally, the R. albus genome sequencing project, led by the PI. Morrison, has supported our identification of many dockerin containing proteins. However, the identification of gene(s) encoding a scaffoldin has been more elusive, and recombinant proteins encoding candidate cohesin modules are now being used in Israel to verify the existence of dockerin-cohesin interactions and cellulosome production by R. albus. The Israeli partners have also conducted virtually all of the studies specific to the second Objective of the proposal. Comparative blotting studies have been conducted using specific antibodies prepare against purified recombinant cohesins and X-domains, derived from cellulosomal scaffoldins of R. flavefaciens 17, a Clostridium thermocellum mutant-preabsorbed antibody preparation, or against CbpC (fimbrial protein) of R. albus 8. The data also suggest that additional cellulolytic bacteria including Fibrobacter succinogenes S85, F. intestinalis DR7 and Butyrivibrio fibrisolvens Dl may also employ cellulosomal modules similar to those of R. flavefaciens 17. Collectively, our work during the grant period has shown that R. albus and other ruminal bacteria employ several novel mechanisms for their adhesion to plant surfaces, and produce both cellulosomal and non-cellulosomal forms of glycoside hydrolases underpinning plant fiber degradation. These improvements in our mechanistic understanding of bacterial adhesion and enzyme regulation now offers the potential to: i) optimize ruminal and hindgut conditions by dietary additives to maximize fiber degradation (e.g. by the addition of select enzymes or PAA/PPA); ii) identify plant-borne influences on adhesion and fiber-degradation, which might be overcome (or improved) by conventional breeding or transgenic plant technologies and; iii) engineer or select microbes with improved adhesion capabilities, cellulosome assembly and fiber degradation. The potential benefits associated with this research proposal are likely to be realized in the medium term (5-10 years).
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Landau, Sergei Yan, John W. Walker, Avi Perevolotsky, Eugene D. Ungar, Butch Taylor, and Daniel Waldron. Goats for maximal efficacy of brush control. United States Department of Agriculture, March 2008. http://dx.doi.org/10.32747/2008.7587731.bard.
Full textAbstract:
Background. Brush encroachment constitutes a serious problem in both Texas and Israel. We addressed the issue of efficacy of livestock herbivory - in the form of goat browsing - to change the ecological balance to the detriment of the shrub vegetation. Shrub consumption by goats is kept low by plant chemical defenses such as tannins and terpenes. Scientists at TAES and ARO have developed an innovative, cost-effective methodology using fecal Near Infrared Spectrometry to elucidate the dietary percentage of targeted, browse species (terpene-richredberry and blueberry juniper in the US, and tannin-rich Pistacialentiscus in Israel) for a large number of animals. The original research objectives of this project were: 1. to clarify the relative preference of goat breeds and the individual variation of goats within breeds, when consuming targeted brush species; 2. to assess the heritability of browse intake and validate the concept of breeding goat lines that exhibit high preference for chemically defended brush, using juniper as a model; 3. to clarify the relative contributions of genetics and learning on the preference for target species; 4. to identify mechanisms that are associated with greater intake of brush from the two target species; 5. to establish when the target species are the most vulnerable to grazing. (Issue no.5 was addressed only partly.) Major conclusions, solutions, achievements: Both the Israel and US scientists put significant efforts into improving and validating the technique of Fecal NIRS for predicting the botanical composition of goat diets. Israeli scientists validated the use of observational data for calibrating fecal NIRS, while US scientists established that calibrations could be used across animals differing in breed and age but that caution should be used in making comparisons between different sexes. These findings are important because the ability to select goat breeds or individuals within a breed for maximal efficiency of brush control is dependent upon accurate measurement of the botanical composition of the diet. In Israel it was found that Damascus goats consume diets more than twice richer in P. lentiscus than Mamber or Boer goats. In the US no differences were found between Angora and Boer cross goats but significant differences were found between individuals within breeds in juniper dietary percentage. In both countries, intervention strategies were found that further increased the consumption of the chemically defended plant. In Israel feeding polyethylene glycol (PEG, MW 4,000) that forms high-affinity complexes with tannins increased P. lentiscus dietary percentage an average of 7 percentage units. In the US feeding a protein supplement, which enhances rates of P450-catalyzed oxidations and therefore the rate of oxidation of monoterpenes, increased juniper consumption 5 percentage units. However, the effects of these interventions were not as large as breed or individual animal effects. Also, in a wide array of competitive tannin-binding assays in Israel with trypsin, salivary proteins did not bind more tannic acid or quebracho tannin than non-specific bovine serum albumin, parotid saliva did not bind more tannins than mixed saliva, no response of tannin-binding was found to levels of dietary tannins, and the breed effect was of minor importance, if any. These fundings strongly suggest that salivary proteins are not the first line of defense from tannin astringency in goats. In the US relatively low values for heritability and repeatability for juniper consumption were found (13% and 30%, respectively), possibly resulting from sampling error or non-genetic transfer of foraging behavior, i.e., social learning. Both alternatives seem to be true as significant variation between sequential observations were noted on the same animal and cross fostering studies conducted in Israel demonstrated that kids raised by Mamber goats showed lower propensity to consume P. lentiscus than counterparts raised by Damascus goats.