Tetrameric mouse acetylcholinesterase: continuum diffusion rate calculations by solving the steady-state Smoluchowski equation using finite element methods

The tetramer is the most important form for acetylcholinesterase in physiological conditions, i.e., in the neuromuscular junction and the nervous system. It is important to study the diffusion of acetylcholine to the active sites of the tetrameric enzyme to understand the overall signal transduction process in these cellular components. Crystallographic studies revealed two different forms of tetramers, suggesting a flexible tetramer model for acetylcholinesterase. Using a recently developed finite element solver for the steady-state Smoluchowski equation, we have calculated the reaction rate for three mouse acetylcholinesterase tetramers using these two crystal structures and an intermediate structure as templates. Our results show that the reaction rates differ for different individual active sites in the compact tetramer crystal structure, and the rates are similar for different individual active sites in the other crystal structure and the intermediate structure. In the limit of zero salt, the reaction rates per active site for the tetramers are the same as that for the monomer, whereas at higher ionic strength, the rates per active site for the tetramers are approximately 67%-75% of the rate for the monomer. By analyzing the effect of electrostatic forces on ACh diffusion, we find that electrostatic forces play an even more important role for the tetramers than for the monomer. This study also shows that the finite element solver is well suited for solving the diffusion problem within complicated geometries.     

Modeling the adoption of innovations in the presence of geographic and media influences

While there is a large body of work examining the effects of social network structure on innovation adoption, models to date have lacked considerations of real geography or mass media. In this article, we show these features are crucial to making more accurate predictions of a social contagion and technology adoption at a city-to-city scale. Using data from the adoption of the popular micro-blogging platform, Twitter, we present a model of adoption on a network that places friendships in real geographic space and exposes individuals to mass media influence. We show that homophily both among individuals with similar propensities to adopt a technology and geographic location is critical to reproducing features of real spatiotemporal adoption. Furthermore, we estimate that mass media was responsible for increasing Twitter's user base two to four fold. To reflect this strength, we extend traditional contagion models to include an endogenous mass media agent that responds to those adopting an innovation as well as influencing agents to adopt themselves.     

Coupled information diffusion--pest dynamics models predict delayed benefits of farmer cooperation in pest management programs

Worldwide, the theory and practice of agricultural extension system have been dominated for almost half a century by Rogers' "diffusion of innovation theory". In particular, the success of integrated pest management (IPM) extension programs depends on the effectiveness of IPM information diffusion from trained farmers to other farmers, an important assumption which underpins funding from development organizations. Here we developed an innovative approach through an agent-based model (ABM) combining social (diffusion theory) and biological (pest population dynamics) models to study the role of cooperation among small-scale farmers to share IPM information for controlling an invasive pest. The model was implemented with field data, including learning processes and control efficiency, from large scale surveys in the Ecuadorian Andes. Our results predict that although cooperation had short-term costs for individual farmers, it paid in the long run as it decreased pest infestation at the community scale. However, the slow learning process placed restrictions on the knowledge that could be generated within farmer communities over time, giving rise to natural lags in IPM diffusion and applications. We further showed that if individuals learn from others about the benefits of early prevention of new pests, then educational effort may have a sustainable long-run impact. Consistent with models of information diffusion theory, our results demonstrate how an integrated approach combining ecological and social systems would help better predict the success of IPM programs. This approach has potential beyond pest management as it could be applied to any resource management program seeking to spread innovations across populations.     

转基因抗虫棉在中国的推广和传播途径研究

转基因抗虫棉是中国商业化应用最成功的转基因作物,已有许多研究对转基因棉花种植的成本效益和农户生产决策的影响因素进行了深入分析,但对其具体推广过程缺乏足够了解。通过小组访谈和创新树的分析方法,以转基因抗虫棉为例,对转基因生物技术在我国的推广和传播途径开展了研究。研究发现,种业公司转基因作物种子的生产能力直接影响转基因作物的初始规模,来自政府研究机构和种业公司的技术推广者在转基因生物技术的扩散过程中都起着重要作用,公共农技推广服务对于宣传相关信息和知识尤为重要,社会资本也有助于转基因抗虫棉在中国的快速传播和采用。研究结论对推进我国公共农技推广体系改革、完善多元社会化服务主体协作及生物技术研发具有重要启示作用。     

DNA and nucleosomes direct distinct folding of a linker histone H1 C-terminal domain

We previously documented condensation of the H1 CTD consistent with adoption of a defined structure upon nucleosome binding using a bulk FRET assay, supporting proposals that the CTD behaves as an intrinsically disordered domain. In the present study, by determining the distances between two different pairs of sites in the C-terminal domain of full length H1 by FRET, we confirm that nucleosome binding directs folding of the disordered H1 C-terminal domain and provide additional distance constraints for the condensed state. In contrast to nucleosomes, FRET observed upon H1 binding to naked DNA fragments includes both intra- and inter-molecular resonance energy transfer. By eliminating inter-molecular transfer, we find that CTD condensation induced upon H1-binding naked DNA is distinct from that induced by nucleosomes. Moreover, analysis of fluorescence quenching indicates that H1 residues at either end of the CTD experience distinct environments when bound to nucleosomes, and suggest that the penultimate residue in the CTD (K195) is juxtaposed between the two linker DNA helices, proposed to form a stem structure in the H1-bound nucleosome.     

Improving Parameter Inference from FRAP Data: an Analysis Motivated by Pattern Formation in the <Emphasis Type="BoldItalic">Drosophila</Emphasis> Wing Disc

Fluorescence recovery after photobleaching (FRAP) is used to obtain quantitative information about molecular diffusion and binding kinetics at both cell and tissue levels of organization. FRAP models have been proposed to estimate the diffusion coefficients and binding kinetic parameters of species for a variety of biological systems and experimental settings. However, it is not clear what the connection among the diverse parameter estimates from different models of the same system is, whether the assumptions made in the model are appropriate, and what the qualities of the estimates are. Here we propose a new approach to investigate the discrepancies between parameters estimated from different models. We use a theoretical model to simulate the dynamics of a FRAP experiment and generate the data that are used in various recovery models to estimate the corresponding parameters. By postulating a recovery model identical to the theoretical model, we first establish that the appropriate choice of observation time can significantly improve the quality of estimates, especially when the diffusion and binding kinetics are not well balanced, in a sense made precise later. Secondly, we find that changing the balance between diffusion and binding kinetics by changing the size of the bleaching region, which gives rise to different FRAP curves, provides a priori knowledge of diffusion and binding kinetics, which is important for model formulation. We also show that the use of the spatial information in FRAP provides better parameter estimation. By varying the recovery model from a fixed theoretical model, we show that a simplified recovery model can adequately describe the FRAP process in some circumstances and establish the relationship between parameters in the theoretical model and those in the recovery model. We then analyze an example in which the data are generated with a model of intermediate complexity and the parameters are estimated using models of greater or less complexity, and show how sensitivity analysis can be used to improve FRAP model formulation. Lastly, we show how sophisticated global sensitivity analysis can be used to detect over-fitting when using a model that is too complex.     

Unveiling the role of Dps in the organization of mycobacterial nucleoid

In order to preserve genetic information in stress conditions, bacterial DNA is organized into higher order nucleoid structure. In this paper, with the help of Atomic Force Microscopy, we show the different structural changes in mycobacterial nucleoid at different points of growth in the presence of different concentrations of glucose in the medium. We also observe that in Mycobacterium smegmatis, two different Dps proteins (Dps1 and Dps2) promote two types of nucleoid organizations. At the late stationary phase, under low glucose availability, Dps1 binds to DNA to form a very stable toroid structure. On the other hand, under the same condition, Dps2-DNA complex forms an incompletely condensed toroid and finally forms a further stable coral reef structure in the presence of RNA. This coral reef structure is stable in high concentration of bivalent ion like Mg(2+).     

Unraveling the impact of lipid domains on the dimerization processes of single-molecule EGFRs of live cells

Epidermal growth factor receptor (EGFR/ErbB1) is a transmembrane protein that can drive cell growth and survival via the ligand-induced dimerization of receptors. Because dimerization is a common mechanism for signal transduction, it is important to improve our understanding of how the dimerization process and membrane structure regulate signal transduction. In this study, we examined the effect of lipid nanodomains on the dimerization process of EGFR molecules. We discovered that after ligand binding, EGFR molecules may move into lipid nanodomains. The lipid nanodomains surrounding two liganded EGFRs can merge during their correlated motion. The transition rates between different diffusion states of liganded EGFR molecules are regulated by the lipid domains. Our method successfully captures both the sensitivity of single-molecule processes and statistic accuracy of data analysis, providing insight into the connection between the mobile clustering process of receptors and the hierarchical structure of plasma membrane.     

A moving boundary model of acrosomal elongation

A sperm penetrates an egg by extending a long, actin-filled tube known as the acrosomal process. This simple example of biomotility is one of the most dramatic. In Thyone, a 90 m process can extend in less than 10 s. Experiments have shown that actin monomers stored in the base of the sperm are transported to the growing tip of the acrosomal process where they add to the ends of the existing filaments.The force that drives the elongation of the acrosomal process has not yet been identified although the most frequently discussed candidate is the actin polymerization reaction. Developing what we believe are realistic moving boundary models of diffusion limited actin fiber polymerization, we show that actin filament growth occurs too slowly to drive acrosomal elongation. We thus believe that other forces, such as osmotically driven water flow, must play an important role in causing the elongation. We conjecture that actin polymerization merely follows to give the appropriate shape to the growing structure and to stabilize the structure once water flow ceases.Work partially supported by the United States Department of Energy     

Social Network Analysis for Program Implementation

This paper introduces the use of social network analysis theory and tools for implementation research. The social network perspective is useful for understanding, monitoring, influencing, or evaluating the implementation process when programs, policies, practices, or principles are designed and scaled up or adapted to different settings. We briefly describe common barriers to implementation success and relate them to the social networks of implementation stakeholders. We introduce a few simple measures commonly used in social network analysis and discuss how these measures can be used in program implementation. Using the four stage model of program implementation (exploration, adoption, implementation, and sustainment) proposed by Aarons and colleagues [1] and our experience in developing multi-sector partnerships involving community leaders, organizations, practitioners, and researchers, we show how network measures can be used at each stage to monitor, intervene, and improve the implementation process. Examples are provided to illustrate these concepts. We conclude with expected benefits and challenges associated with this approach.     

Adaptability and the integration of computer-based information processing into the dynamics of organizations

The structure of a system influences its adaptability. An important result of adaptability theory is that subsystem independence increases adaptability [Conrad, M., 1983. Adaptability. Plenum Press, New York]. Adaptability is essential in systems that face an uncertain environment such as biological systems and organizations. Modern organizations are the product of human design. And so it is their structure and the effect that it has on their adaptability. In this paper we explore the potential effects of computer-based information processing on the adaptability of organizations. The integration of computer-based processes into the dynamics of the functions they support and the effect it has on subsystem independence are especially relevant to our analysis.     

Faster Is More Different: Mean-Field Dynamics of Innovation Diffusion

Based on a recent model of paradigm shifts by Bornholdt et al., we studied mean-field opinion dynamics in an infinite population where an infinite number of ideas compete simultaneously with their values publicly known. We found that a highly innovative society is not characterized by heavy concentration in highly valued ideas: Rather, ideas are more broadly distributed in a more innovative society with faster progress, provided that the rate of adoption is constant, which suggests a positive correlation between innovation and technological disparity. Furthermore, the distribution is generally skewed in such a way that the fraction of innovators is substantially smaller than has been believed in conventional innovation-diffusion theory based on normality. Thus, the typical adoption pattern is predicted to be asymmetric with slow saturation in the ideal situation, which is compared with empirical data sets.     

Fluorescence correlation spectroscopy diffusion laws to probe the submicron cell membrane organization

To probe the complexity of the cell membrane organization and dynamics, it is important to obtain simple physical observables from experiments on live cells. Here we show that fluorescence correlation spectroscopy (FCS) measurements at different spatial scales enable distinguishing between different submicron confinement models. By plotting the diffusion time versus the transverse area of the confocal volume, we introduce the so-called FCS diffusion law, which is the key concept throughout this article. First, we report experimental FCS diffusion laws for two membrane constituents, which are respectively a putative raft marker and a cytoskeleton-hindered transmembrane protein. We find that these two constituents exhibit very distinct behaviors. To understand these results, we propose different models, which account for the diffusion of molecules either in a membrane comprising isolated microdomains or in a meshwork. By simulating FCS experiments for these two types of organization, we obtain FCS diffusion laws in agreement with our experimental observations. We also demonstrate that simple observables derived from these FCS diffusion laws are strongly related to confinement parameters such as the partition of molecules in microdomains and the average confinement time of molecules in a microdomain or a single mesh of a meshwork.     

How to Control the Molecular Architecture of a Monolayer of Proteins Supported by a Lipid Bilayer

In this work, we report the spontaneous formation of a new structure composed of two lipid layers surrounding a dense monolayer of soluble proteins (lysozyme). We extend a process, initially discovered with nonionic surfactants to phospholipids (DMPC and DOPC). The motor of the protein insertion process is the difference between the protein chemical potential in the solution and in the freshly formed Newton black film (NBF). This process is completely controlled by adjusting the protein chemical potential in the solution. By means of x-ray reflectivity, we follow the evolution of the freestanding sandwich structure until a stable equilibrium state is reached. Depending on the lipid concentration with respect to the protein concentration, we observe two different behaviors of the film leading to the formation of such unique structure: at the highest lipid concentration, the usual protein diffusion into the NBF, and, at the lowest lipid concentration, the spontaneous formation of a sandwich structure immediately obtained after the drainage. Finally, we show that the insertion process is reversible, because, if the lipid concentration varies in the bulk solution, a “deswelling” of the film can be observed.     

Exploring pathways and barriers for coupled ET/PT in cytochrome c oxidase: a general framework for examining energetics and mechanistic alternatives

Gaining a detailed understanding of the energetics of the proton pumping process in cytochrome c oxidase (CcO) is one of the challenges of modern biophysics. Although there are several current mechanistic proposals, most of these ideas have not been subjected to consistent structure-function considerations. In particular most works have not related the activation barriers for different mechanistic proposals to the protein structure. The present work describes a general approach for exploring the energetics of different feasible models of the action of CcO, using the observed protein structure, established simulation methods and a modified Marcus' formulation. We start by reviewing our methods for evaluation of the energy diagrams for different proton translocation paths and then present a systematic analysis of various constraints that should be imposed on any energy diagram for the pumping process. After the general analysis we turn to the actual computational study, where we construct energy diagrams for forward and backward paths, using the estimated calculated reduction potentials and pK(a) values of all the relevant sites (including internal water molecules). We then explore the relationship between the calculated energy diagrams and key experimental constraints. This comparison allows us to identify some barriers that are not fully consistent with the overall requirement for an efficient pumping. In particular we identify back leakage channels, which are hard to block without stopping the forward channels. This helps to identify open problems that will require further experimental and theoretical studies. We also consider reasonable adjustments of the calculated barriers that may lead to a working pump. Although the present analysis does not establish a unique and workable model for the mechanism of CcO, it presents what is probably the most consistent current analysis of the barriers for different feasible pathways. Perhaps more importantly, the framework developed here should provide a general way for examining any proposal for the action of CcO as well as for the analysis of further experimental findings about the action of this fascinating system.     

Exploring pathways and barriers for coupled ET/PT in cytochrome c oxidase: A general framework for examining energetics and mechanistic alternatives

Gaining a detailed understanding of the energetics of the proton pumping process in cytochrome c oxidase (CcO) is one of the challenges of modern biophysics. Although there are several current mechanistic proposals, most of these ideas have not been subjected to consistent structure-function considerations. In particular most works have not related the activation barriers for different mechanistic proposals to the protein structure. The present work describes a general approach for exploring the energetics of different feasible models of the action of CcO, using the observed protein structure, established simulation methods and a modified Marcus' formulation. We start by reviewing our methods for evaluation of the energy diagrams for different proton translocation paths and then present a systematic analysis of various constraints that should be imposed on any energy diagram for the pumping process. After the general analysis we turn to the actual computational study, where we construct energy diagrams for forward and backward paths, using the estimated calculated reduction potentials and pK_a values of all the relevant sites (including internal water molecules). We then explore the relationship between the calculated energy diagrams and key experimental constraints. This comparison allows us to identify some barriers that are not fully consistent with the overall requirement for an efficient pumping. In particular we identify back leakage channels, which are hard to block without stopping the forward channels. This helps to identify open problems that will require further experimental and theoretical studies. We also consider reasonable adjustments of the calculated barriers that may lead to a working pump. Although the present analysis does not establish a unique and workable model for the mechanism of CcO, it presents what is probably the most consistent current analysis of the barriers for different feasible pathways. Perhaps more importantly, the framework developed here should provide a general way for examining any proposal for the action of CcO as well as for the analysis of further experimental findings about the action of this fascinating system.     

Using metadynamics to understand the mechanism of calmodulin/target recognition at atomic detail

The ability of calcium-bound calmodulin (CaM) to recognize most of its target peptides is caused by its binding to two hydrophobic residues ('anchors'). In most of the CaM complexes, the anchors pack against the hydrophobic pockets of the CaM domains and are surrounded by fully conserved Met side chains. Here, by using metadynamics simulations, we investigate quantitatively the energetics of the final step of this process using the M13 peptide, which has a high affinity and spans the sequence of the skeletal myosin light chain kinase, an important natural CaM target. We established the accuracy of our calculations by a comparison between calculated and NMR-derived structural and dynamical properties. Our calculations provide novel insights into the mechanism of protein/peptide recognition: we show that the process is associated with a free energy gain similar to that experimentally measured for the CaM complex with the homologous smooth muscle MLCK peptide (Ehrhardt et al., 1995, Biochemistry 34, 2731). We suggest that binding is dominated by the entropic effect, in agreement with previous proposals. Furthermore, we explain the role of conserved methionines by showing that the large flexibility of these side chains is a key feature of the binding mechanism. Finally, we provide a rationale for the experimental observation that in all CaM complexes the C-terminal domain seems to be hierarchically more important in establishing the interaction.     

Evolving patterns in a collaboration network of global R&D on monoclonal antibodies

We investigated the evolution process of collaborative inter-organizational network of the research and development (R&D) on monoclonal antibody (mAb) over the past 30 y. The annual detection of the collaboration network provides dynamics on network structures and relationship changes among different organizations. Our research showed continuous growth of the network's scale and complexity due to the constant entry of new organizations and the forging of new partnering relationships. The evolving topological features reveal a core-periphery structure that became clearer over time and an increasing heterogeneity within the collaborative mAb R&D network. As measured by the number of network participants, dedicated biotechnology firms (DBFs) were the dominant organization form in the field of mAb development, but their average centrality was reduced during the period of 2004–2009, when pharmaceutical companies took over the positions of DBFs. Along with the network evolution, 2 waves of substitution on the leading positions were driven by technological innovations and mergers and acquisitions (M&A). In addition, this study analyzed organizational-level behaviors to help understand the evolution of network structures over the field of mAb development across the different technologically innovative or economic contexts.     

Social Capital and Sustainable Coffee Certifications in Costa Rica

Social capital has many applications in the adoption and management of voluntary certifications such as Fair trade, Rainforest Alliance, Utz, and CAFE Practices. We used a mixed-methods approach to assess social capital in five Costa Rican coffee cooperatives. We applied this information to its effects on the management of sustainable coffee certifications We found that the level of social capital affects the manner in which cooperatives manage certifications in terms of incentivizing certified members, distributing the profits from certification, and cooperating with outside organizations. Generalized trust was found to have an important link with voluntary participation in Rainforest Alliance certification when no financial incentive was provided. However, given the small differences among the cooperatives in micro-scale social capital, we conclude that certifications have not or have not yet made great impacts on the social capital on the micro-scale. This research presents important considerations for employing certifications and other sustainable development projects in different national contexts.