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The crystal structure of the dopamine D3 receptor (D3R) in complex with eticlopride inspired the design of bitopic ligands that explored (1) N-alkylation of the eticlopride's pyrrolidine ring, (2) shifting of the position of the pyrrolidine nitrogen, (3) expansion of the pyrrolidine ring system, and (4) incorporation of O-alkylations at the 4-position. Structure activity relationships (SAR) revealed that moving the N- or expanding the pyrrolidine ring was detrimental to D2R/D3R binding affinities. Small pyrrolidine N-alkyl groups were poorly tolerated, but the addition of a linker and secondary pharmacophore (SP) improved affinities. Moreover, O-alkylated analogues showed higher binding affinities compared to analogously N-alkylated compounds, e.g., O-alkylated 33 (D3R, 0.436 nM and D2R, 1.77 nM) vs the N-alkylated 11 (D3R, 6.97 nM and D2R, 25.3 nM). All lead molecules were functional D2R/D3R antagonists. Molecular models confirmed that 4-position modifications would be well-tolerated for future D2R/D3R bioconjugate tools that require long linkers and or sterically bulky groups.

We have developed and characterized a novel D2R antagonist with exceptional GPCR selectivity – ML321. In functional profiling screens of 168 different GPCRs, ML321 showed little activity beyond potent inhibition of the D2R, and to a lesser extent the D3R, demonstrating excellent receptor selectivity. The D2R selectivity of ML321 may be related to the fact that, unlike other monoaminergic ligands, ML321 lacks a positively charged amine group and adopts a unique binding pose within the orthosteric binding site of the D2R. PET imaging studies in non-human primates demonstrated that ML321 penetrates the CNS and occupies the D2R in a dose-dependent manner. Behavioral paradigms in rats demonstrate that ML321 can selectively antagonize a D2R-mediated response (hypothermia) while not affecting a D3R-mediated response (yawning) using the same dose of drug, thus indicating exceptionalin vivoselectivity. We also investigated the effects of ML321 in animal models that are predictive of antipsychotic efficacy in humans. We found that ML321 attenuates both amphetamine- and phencyclidine-induced locomotor activity and restored pre-pulse inhibition (PPI) of acoustic startle in a dose-dependent manner. Surprisingly, using doses that were maximally effective in both the locomotor and PPI studies, ML321 was relatively ineffective in promoting catalepsy. Kinetic studies revealed that ML321 exhibits slow-on and fast-off receptor binding rates, similar to those observed with atypical antipsychotics with reduced extrapyramidal side effects. Taken together, these observations suggest that ML321, or a derivative thereof, may exhibit “atypical” antipsychotic activity in humans with significantly fewer side effects than observed with currently FDA-approved D2R antagonists.

Psychostimulant drugs, such as cocaine, inhibit dopamine reuptake via blockading the dopamine transporter (DAT), which is the primary mechanism underpinning their abuse. Atypical DAT inhibitors are dissimilar to cocaine and can block cocaine- or methamphetamine-induced behaviors, supporting their development as part of a treatment regimen for psychostimulant use disorders. When developing these atypical DAT inhibitors as medications, it is necessary to avoid off-target binding that can produce unwanted side effects or toxicities. In particular, the blockade of a potassium channel, human ether-a-go-go (hERG), can lead to potentially lethal ventricular tachycardia. In this study, we established a counter screening platform for DAT and against hERG binding by combining machine learning-based quantitative structure-activity relationship (QSAR) modeling, experimental validation, and molecular modeling and simulations. Our results show that the available data are adequate to establish robust QSAR models, as validated by chemical synthesis and pharmacological evaluation of a validation set of DAT inhibitors. Furthermore, the QSAR models based on subsets of the data according to experimental approaches used have predictive power as well, which opens the door to target specific functional states of a protein. Complementarily, our molecular modeling and simulations identified the structural elements responsible for a pair of DAT inhibitors having opposite binding affinity trends at DAT and hERG, which can be leveraged for rational optimization of lead atypical DAT inhibitors with desired pharmacological properties.

Adhesion G protein-coupled receptor latrophilin 3 (ADGRL3), a cell adhesion molecule highly expressed in the central nervous system, acts in synapse formation through trans interactions with its ligands. It is largely unknown if these interactions serve a purely adhesive function or can modulate G protein signaling. To assess how different structural elements of ADGRL3 (e.g., the adhesive domains, autoproteolytic cleavage site, or tethered agonist (TA)) impact receptor function, we require constructs that disrupt specific receptor features without impacting others. While we showed previously that mutating conserved Phe and Met residues in the TA of ADGRL3-C-terminal fragment (CTF), a CTF truncated to the G protein-coupled receptor proteolysis site, abolishes receptor-mediated G protein activation, we now find that autoproteolytic cleavage is disrupted in the full-length version of this construct. To identify a construct that disrupts TA-dependent activity without impacting proteolysis, we explored other mutations in the TA. We found that mutating the sixth and seventh residues of the TA, Leu and Met, to Ala impaired activity in a serum response element activity assay for both full-length and CTF constructs. We confirmed this activity loss results from impaired G protein coupling using an assay that acutely exposes the TA through controlled proteolysis. The ADGRL3 mutant expresses normally at the cell surface, and immunoblotting shows that it undergoes normal autoproteolysis. Thus, we found a construct that disrupts tethered agonism while retaining autoproteolytic cleavage, providing a tool to disentangle these functions in vivo. Our approach and specific findings are likely to be broadly applicable to other adhesion receptors.

As asynchronous online discussions gain a broader usage due to the COVID-19 epidemic, the need for understanding of students' ideas from unstructured textual data becomes more pressing. In this experimental study, we examine the effects of recommendations on message quality and community formation from voluminous online discussions. Drawing on literature from group cognition, knowledge building discourse, and learning analytics, we calculate message quasi-quality index (QQI) scores based on message lexical complexity and topic-related keyword usage by participants in explaining their ideas. Furthermore, we examine the empirical evidence on the relationship between QQI scores and participants' interactions. Finally, we visualize network structures via sociograms and hierarchically cluster participants to identify subgroups. Our analysis of 281 messages finds that recommendations helped participants to write more messages that compared alternative viewpoints and refined preliminary ideas with higher QQI scores. Results also show that recommendations cultivated a sense of collective agency to increase the opportunity for creativity and reduce the likelihood that peripheral participants will be dissatisfied and fail to identify with a community. Theoretical contributions and practical implications are discussed.

Online Word-of-Mouth communication, or eWOM, has dramatically changed the way people network, interact, and share knowledge. Studies have examined why consumers choose to share knowledge online, especially online product reviews, as well as the motivations of individuals to share product ideas online. However, the role of gender in shaping the motivation and types of knowledge shared online has been given little consideration. Using concepts from Social Exchange Theory and the Theory of Reasoned Action, we address this research gap by developing and testing a model of gender’s influence on knowledge sharing in a social media context. A PLS analysis of survey data from 257 students indicates that reputation, altruism, and subjective norms are key motivators for knowledge sharing intention in social media. More importantly, that gender plays a moderating role within the motivation-knowledge sharing relationship. We also find that subjective norms have a greater impact on knowledge sharing with women than with men. Collectively, our research results highlight individualized factors for improving customer participation in external facing social media for marketing and product innovation.

International audience; The temperature dependence of glass relaxation has been intensively studied; however, the effect of an imposed pressure history on relaxation behavior is poorly understood. In this study, we subjected SCHOTT N-BK7® borosilicate glasses to isostatic compression in a Paterson press (PP) and a gas pressure chamber (GPC). The pressure ranged from 0.1 GPa to 2 GPa for various dwell temperatures and times near the glass transition region. Comparison with our recent results on the same glass using the piston-cylinder apparatus (PC, 0.5-1.5 GPa) reveals that the density of a glass, which has been quenched from the equilibrium state under high pressure at 2 K/min (pressure quench), increases approximately linearly with increasing pressure up to 2 GPa. Considering the volume recovery results at ambient pressure, we assert that the preceding high-pressure treatment in PC (uniaxial loading) generates a similar isostatic pressure effect on N-BK7 glass as those of PP and GPC treatments. Finally, we verify the previously proposed two-internal-parameter relaxation model on the volume recovery data using the three different compression methods. With a new set of parameters in the model, we can account for the pressure and temperature dependence of volume relaxation even for the samples quenched from nonequilibrium states at high pressure.

Vaccines are regarded as the most cost-effective countermeasure against infectious diseases. One challenge often affecting vaccine development is antigenic diversity or pathogen heterogeneity. Different strains produce immunologically heterogeneous virulence factors, therefore an effective vaccine needs to induce broad-spectrum host immunity to provide cross-protection. Recent advances in genomics and proteomics, particularly computational biology and structural biology, establishes structural vaccinology and highlights the feasibility of developing effective and precision vaccines. Here, we introduce the epitope- and structure-based vaccinology platform multiepitope-fusion-antigen (MEFA), and provide instructions to generate polyvalent MEFA immunogens for vaccine development. Conceptually, MEFA combines epitope vaccinology and structural vaccinology to enable a protein immunogen to present heterogeneous antigenic domains (epitopes) and to induce broadly protective immunity against different virulence factors, strains or diseases. Methodologically, the MEFA platform first identifies a safe, structurally stable and strongly immunogenic backbone protein and immunodominant (ideally neutralizing or protective) epitopes from heterogeneous strains or virulence factors of interest. Then, assisted with protein modeling and molecule dynamic simulation, MEFA integrates heterogeneous epitopes into a backbone protein via epitope substitution for a polyvalent MEFA protein and mimics epitope native antigenicity. Finally, the MEFA protein is examined for broad immunogenicity in animal immunization, and assessed for potential application for multivalent vaccine development in preclinical studies.

The crystal structure of the dopamine D(3) receptor (D(3)R) in complex with eticlopride inspired the design of bitopic ligands that explored 1) N-alkylation of eticlopride’s pyrrolidine ring, 2) shifting the position of the pyrrolidine nitrogen, 3) expanding the pyrrolidine ring system, and 4) incorporating O-alkylations at the 4-position. Structure Activity Relationships (SAR) revealed that moving the N- or expanding the pyrrolidine ring was detrimental to D(2)R/D(3)R binding affinities. Small pyrrolidine N-alkyl groups were poorly tolerated, but addition of a linker and secondary pharmacophore (SP) improved affinities. Moreover, O-alkylated analogues showed higher binding affinities compared to analogously N-alkylated compounds, e.g., O-alkylated 33 (D(3)R, 0.436 nM and D(2)R, 1.77 nM) vs. the N-alkylated 11 (D(3)R, 6.97 nM and D(2)R, 25.3 nM). All lead molecules were functional D(2)R/D(3)R antagonists. Molecular models confirmed that 4-position modifications would be well-tolerated for future D(2)R/D(3)R bioconjugate tools that require long linkers and or sterically bulky groups.

In the context of large annotation-based literature discussions, this research examines the affordances of recommender systems on users’ disorientation. Drawing insights from literature on group co...

Borosilicate glasses have been in widespread use for over a century; however, a detailed understanding of the structural response to densification is still lacking. In this work, two commercial borosilicate glasses, viz., SCHOTT N-BK7® (N-BK7) and Borofloat33® (Boro33), are hot compressed up to 2 GPa with nitrogen gas, and the structural response to this densification is explored via 11B solid-state nuclear magnetic resonance (NMR) spectroscopy and classical molecular dynamics (MD) simulations. The molar volume (Vm) of N-BK7 and Boro33 decreases ~5% and ~10%, respectively, as a result of hot compression at 2 GPa. The NMR results demonstrate the presence of three different types of fourfold coordinated boron species (B[4]), which are confirmed in MD simulations to be (Formula presented.) (0B,4Si[4]), (Formula presented.) (1B[3],3Si[4]), and (Formula presented.) (1B[4],3Si[4]) (where subscripts represent different B[4] types and brackets indicate the next nearest neighbors (NNN)). The NMR results also show that the fraction of B[4] increases by ~13% in N-BK7 glass upon hot compression at 2 GPa via the trigonal boron to tetrahedral boron (B[3] to B[4]) conversion, while the fraction of B[4] in Boro33 glass only increases by ~2% at the same pressure, despite the fact that the Vm decrease in N-BK7 is double that of Boro33. The MD simulations capture the experimental trends in B[4] populations, despite an underestimation of the B[4] increase of N-BK7 (only ~6%) at 2 GPa. Moreover, the MD simulations suggest that the Vm reduction is a linear function of bond angle change and the fraction of Si-O-Si and B[4]-O-Si. The modifiers and boron coordination conversion also influence the volume densification of borosilicate glasses by increasing the difficulty of bond bending, decreasing the bond lengths, and increasing the population of B[4]-O-Si linkages. Finally, the B[4] to B[4] conversion, that is, (Formula presented.) (1B[3],3Si[4]) and (Formula presented.) (1B[4],3Si[4]) to (Formula presented.) (0B,4Si[4]), is observed in hot compressed N-BK7 and Boro33 from NMR and qualitatively confirmed in MD.

Collagen prolyl 4-hydroxylase 1 (C-P4H1) is an &alpha, ketoglutarate (&alpha, KG)-dependent dioxygenase that catalyzes 4-hydroxylation of proline on collagen. C-P4H1-induced prolyl hydroxylation is required for proper collagen deposition and cancer metastasis. Therefore, targeting C-P4H1 is considered a potential therapeutic strategy for collagen-related cancer progression and metastasis. However, no C-P4H1 inhibitors are available for clinical testing, and the high content assay is currently not available for C-P4H1 inhibitor screening. In the present study, we developed a high-throughput screening assay by quantifying succinate, a byproduct of C-P4H-catalyzed hydroxylation. C-P4H1 is the major isoform of collagen prolyl 4-hydroxylases (CP4Hs) that contributes the majority prolyl 4-hydroxylase activity. Using C-P4H1 tetramer purified from the eukaryotic expression system, we showed that the Succinate-GloTM Hydroxylase assay was more sensitive for measuring C-P4H1 activity compared with the hydroxyproline colorimetric assay. Next, we performed high-throughput screening with the FDA-approved drug library and identified several new C-P4H1 inhibitors, including Silodosin and Ticlopidine. Silodosin and Ticlopidine inhibited C-P4H1 activity in a dose-dependent manner and suppressed collagen secretion and tumor invasion in 3D tissue culture. These C-P4H1 inhibitors provide new agents to test clinical potential of targeting C-P4H1 in suppressing cancer progression and metastasis.

Conformational changes of proteins are essential to their functions. Yet it remains challenging to measure the amplitudes and timescales of protein motions. Here we show that the cytolysin A (ClyA) nanopore was used as a molecular tweezer to trap a single maltose-binding protein (MBP) within its lumen, which allows conformation changes to be monitored as electrical current fluctuations in real time. In contrast to the current two state binding model, the current measurements revealed three distinct ligand-bound states for MBP in the presence of reducing saccharides. Our analysis reveal that these three states represented MBP bound to different isomers of reducing sugars. These findings contribute to on the understanding of the mechanism of substrate recognition by MBP and illustrate that the nanopore tweezer is a powerful, label-free, single-molecule approach for studying protein conformational dynamics under functional conditions.

By analyzing and simulating inactive conformations of the highly homologous dopamine D2 and D3 receptors (D2R and D3R), we find that eticlopride binds D2R in a pose very similar to that in the D3R/eticlopride structure but incompatible with the D2R/risperidone structure. In addition, risperidone occupies a sub-pocket near the Na+ binding site, whereas eticlopride does not. Based on these findings and our experimental results, we propose that the divergent receptor conformations stabilized by Na+-sensitive eticlopride and Na+-insensitive risperidone correspond to different degrees of inverse agonism. Moreover, our simulations reveal that the extracellular loops are highly dynamic, with spontaneous transitions of extracellular loop 2 from the helical conformation in the D2R/risperidone structure to an extended conformation similar to that in the D3R/eticlopride structure. Our results reveal previously unappreciated diversity and dynamics in the inactive conformations of D2R. These findings are critical for rational drug discovery, as limiting a virtual screen to a single conformation will miss relevant ligands., eLife digest Almost a third of prescribed drugs work by acting on a group of proteins known as GPCRs (short for G-protein coupled receptors), which help to transmit messages across the cell’s outer barrier. The neurotransmitter dopamine, for instance, can act in the brain and body by attaching to dopamine receptors, a sub-family of GPCRs. The binding process changes the three-dimensional structure (or conformation) of the receptor from an inactive to active state, triggering a series of molecular events in the cell. However, GPCRs do not have a single ‘on’ or ‘off’ state; they can adopt different active shapes depending on the activating molecule they bind to, and this influences the type of molecular cascade that will take place in the cell. Some evidence also shows that classes of GPCRs can have different inactive structures; whether this is also the case for the dopamine D2 and D3 receptors remained unclear. Mapping out inactive conformations of receptors is important for drug discovery, as compounds called antagonists can bind to inactive receptors and interfere with their activation. Lane et al. proposed that different types of antagonists could prefer specific types of inactive conformations of the dopamine D2 and D3 receptors. Based on the structures of these two receptors, the conformations of D2 bound with the drugs risperidone and eticlopride (two dopamine antagonists) were simulated and compared. The results show that the inactive conformations of D2 were very different when it was bound to eticlopride as opposed to risperidone. In addition D2 and D3 showed a very similar conformation when attached to eticlopride. The two drugs also bound to the inactive receptors in overlapping but different locations. These computational findings, together with experimental validations, suggest that D2 and D3 exist in several inactive states that only allow the binding of specific drugs; these states could also reflect different degrees of inactivation. Overall, the work by Lane et al. contributes to a more refined understanding of the complex conformations of GPCRs, which could be helpful to screen and develop better drugs.

Public health reports indicate that high PM2.5 concentration can impair the respiratory health of the residents, especially for those affected by asthma. Therefore, there is a need to determine the deposition mechanism and efficiencies for PM2.5 in asthmatic human airways. In this study, gas flow dynamics and deposition fractions (DFs) of PM2.5 in generations 10–11 of Weibel's lung model were investigated where the two-phase gas-solid flow behaviors in healthy and asthmatic airways were considered. The gas phase was modeled as a transient laminar and incompressible flow while the discrete phase model (DPM) was applied for the particle phase. Three different air flow rates under rest, light activity, and moderate exercise were considered. For the healthy airways, higher total mass DFs were observed during a moderate exercise as compared to rest and light activity conditions. Deposition fractions were higher in asthmatic airways compared to those of healthy ones, stemming from tapering in the airways as well as complex secondary flow fields, namely, Dean vortices, in the folds. Deposition was mainly due to inertial forces of particles, but a small amount of PM2.5 was deposited near the entrance of asthmatic tube, as a result of the secondary flow. The numerical results revealed that the Dean vortices was an important factor for particle deposition. With increased DF, asthmatic people have a higher total respiratory dose of PM2.5 for a given exposure compared to healthy individuals. Thus contributing to their increased susceptibility to adverse health effects caused by PM2.5.

In this paper, we calculate four different kinds of means—AM, GM, HM, and GDM—to investigate the risk-return contour using Markowitz risk minimization and Sharpe’s angle maximization models. For a given value (target portfolio return), the rank order of risk or variance-covariance (υ) can change. In the vertical segment of an efficient frontier curve, we observed v(GDM) > v(HM) > v(GM) > v(AM). At higher k values, the rank changes to v(GDM) > v(HM) > v(AM) > v(GM). That is to say, ranking a portfolio using different kinds of means may well give different rankings depending on what k value one is evaluating. It is also shown the harmonic mean should not be used in the case of a small negative growth rate in stock prices.

Recasting temperature replica exchange (T-RE) as a special case of Gibbs sampling has led to a simple and efficient scheme for enhanced mixing (Chodera and Shirts, J. Chem. Phys., 2011, 135, 194110). To critically examine if T-RE with independence sampling (T-REis) improves conformational sampling, we performed T-RE and T-REis simulations of ordered and disordered proteins using coarse-grained and atomistic models. The results demonstrate that T-REis effectively increase the replica mobility in temperatures space with minimal computational overhead, especially for folded proteins. However, enhanced mixing does not translate well into improved conformational sampling. The convergences of thermodynamic properties interested are similar, with slight improvements for T-REis of ordered systems. The study re-affirms the efficiency of T-RE does not appear to be limited by temperature diffusion, but by the inherent rates of spontaneous large-scale conformational re-arrangements. Due to its simplicity and efficacy of enhanced mixing, T-REis is expected to be more effective when incorporated with various Hamiltonian-RE protocols. © 2017 Wiley Periodicals, Inc.

Conformational changes of proteins are essential to their functions. Yet it remains challenging to measure the amplitudes and timescales of protein motions. Here we show that the ClyA nanopore can be used as a molecular tweezer to trap a single maltose-binding protein (MBP) within its lumen, which allows conformation changes to be monitored as electrical current fluctuations in real time. The current measurements revealed three distinct ligand-bound states for MBP in the presence of reducing saccharides. Our biochemical and kinetic analysis reveal that these three states represented MBP bound to different isomers of reducing sugars. These findings shed light on the mechanism of substrate recognition by MBP and illustrate that the nanopore tweezer is a powerful, label-free, single-molecule approach for studying protein conformational dynamics under functional conditions.

In this study, we compare two sets of classical interatomic potentials for borosilicate glasses by investigating two major commercial borosilicate glasses, Borofloat®33 (Boro33) and N-BK7®, via molecular dynamics simulations. To evaluate the performance of the two potentials, structural and elastic properties are analyzed for the two glass compositions and are compared with available experimental values. We find that the potential by Wang et al. [M. Wang, N.M. Anoop Krishnan, B. Wang, M.M. Smedskjaer, J.C. Mauro, and M. Bauchy, J. Non. Cryst. Solids, 498 294–304 (2018)] provides a closer N4 value for Boro33 but underpredicts the N4 value for N-BK7. In contrast, the N4 value of N-BK7 using the potential of Deng and Du [L. Deng and J. Du, J. Am. Ceram. Soc., 102 [5] 2482–2505 (2019)] agrees well with the experimental data, but that of Boro33 sample is overpredicted. Our result also indicates that Wang's potential gives a better prediction in the short-range structure, while Du's potential provides a closer medium-range structure compared with the experimental data. Neither set of potentials is able to provide accurate predictions of elastic moduli. Wang's potential predicts lower elastic modulus due to the under predicted N4 value, whereas Du's potential yields higher elastic modulus compared with the experimental values, resulting from its overpredicted N4 value.

A topological constraint model is developed to predict the compositional scaling of glass transition temperature (Tg) in a metal–organic framework glass, agZIF-62 [Zn(Im2–xbImx)]. A hierarchy of bond constraints is established using a combination of experimental results and molecular dynamic simulations with ReaxFF. The model can explain the topological origin of Tg as a function of the benzimidazolate concentration with an error of 3.5 K. The model is further extended to account for the effect of 5-methylbenzimidazolate, enabling calculation of a ternary diagram of Tg with a mixture of three organic ligands in an as-yet unsynthesized, hypothetical framework. We show that topological constraint theory is an effective tool for understanding the properties of metal–organic framework glasses.

We show that by using different x-ray irradiation times of BSA-coated Au nanoparticles (NPs) we can change their ultraviolet-stimulated photoluminescence and shift the spectral weight over the visible spectral range. This is due to the interplay of two emission bands, one due to BSA and the other related to gold. The emission properties did not change with time over a period of several months. (C) 2014 Published by Elsevier B.V.

Vaccine development often encounters the challenge of virulence heterogeneity. Enterotoxigenic Escherichia coli (ETEC) bacteria producing immunologically heterogeneous virulence factors are a leading cause of children’s diarrhea and travelers’ diarrhea. Currently, we do not have licensed vaccines against ETEC bacteria. While conventional methods continue to make progress but encounter challenge, new computational and structure-based approaches are explored to accelerate ETEC vaccine development. In this study, we applied a structural vaccinology concept to construct a structure-based multiepitope fusion antigen (MEFA) to carry representing epitopes of the seven most important ETEC adhesins [CFA/I, CFA/II (CS1-CS3), CFA/IV (CS4-CS6)], simulated antigenic structure of the CFA/I/II/IV MEFA with computational atomistic modeling and simulation, characterized immunogenicity in mouse immunization, and examined the potential of structure-informed vaccine design for ETEC vaccine development. A tag-less recombinant MEFA protein (CFA/I/II/IV MEFA) was effectively expressed and extracted. Molecular dynamics simulations indicated that this MEFA immunogen maintained a stable secondary structure and presented epitopes on the protein surface. Empirical data showed that mice immunized with the tagless CFA/I/II/IV MEFA developed strong antigen-specific antibody responses, and mouse serum antibodies significantly inhibited in vitro adherence of bacteria expressing these seven adhesins. These results revealed congruence of antigen immunogenicity between computational simulation and empirical mouse immunization and indicated this tag-less CFA/I/II/IV MEFA potentially an antigen for a broadly protective ETEC vaccine, suggesting a potential application of MEFA-based structural vaccinology for vaccine design against ETEC and likely other pathogens.

Protein structural stability and biological functionality are dictated by the formation of intradomain cores and interdomain interfaces, but the intricate sequence-structure-function interrelationships in the packing of protein cores and interfaces remain difficult to elucidate due to the intractability of enumerating all packing possibilities and assessing the consequences of all the variations. In this work, groups of β strand residues of model antibody variable domains were randomized with saturated mutagenesis and the functional variants were selected for high-throughput sequencing and high-throughput thermal stability measurements. The results show that the sequence preferences of the intradomain hydrophobic core residues are strikingly flexible among hydrophobic residues, implying that these residues are coupled indirectly with antigen binding through energetic stabilization of the protein structures. By contrast, the interdomain interface residues are directly coupled with antigen binding. The interdomain interface should be treated as an integral part of the antigen-binding site.

Nanoindentation was performed on a metallic glass with an embedded glassy nanoparticle using molecular dynamics simulations to investigate the effect of the second phase on shear band formation and crack initiation. It was found that the addition of a nanoparticle having a lower elastic modulus compared to that of matrix centralizes the region of shear strain below the indenter, which facilitates crack initiation. In contrast, the addition of a nanoparticle with a higher elastic modulus has the effect of branching the shear band, which confines the shear bands within the hydrostatic zone and suppresses crack initiation.

This paper studies the effects of alternative tax policies (a consumption tax, seigniorage tax, and tax switch) on economic growth under different methods of government budget adjustment in a monetary endogenous growth model with a labor–leisure choice and a cash-in-advance constraint which is only imposed on consumption. It is found that the validity of both the Mundell–Tobin effect and the consumption tax neutrality crucially depends on the adjustment methods used to maintain the balanced government budget. In addition, we find that a switch from a consumption tax to a seigniorage tax unambiguously enhances economic growth. This result stands in sharp contrast to that of Ho et al. (J Money Credit Bank 39:105–131, 2007), in that it does confirm the validity of the qualitative equivalence between the money-in-the-utility-function and cash-in-advance approaches in terms of the effect of tax shifting away from a consumption tax towards a seigniorage tax.

Human capital plays an important role and acts as a strategic resource that helps firms achieve strategic competitiveness. In the global business context, understanding the expectations and behaviors of employees in different cultures is paramount for international and multicultural organizations to succeed. This paper examines the task and relationship orientations as well as the work overload stress perception of people in the low-context culture of the Netherlands and in the high-context culture of Vietnam. As a result of the analysis of 396 responses, some significant differences were found between the two samples. It appears that Vietnamese have significantly higher scores on task, relationship and stress orientations than Dutch respondents. While gender is a significant factor in task and relationship orientations, it did not demonstrate any differences in the stress perceptions of these respondents. In this paper, literature on Dutch and Vietnamese cultures is presented along with practical application, suggestions and implications for future studies.

Accurate treatment of solvent environment is critical for reliable simulations of protein conformational equilibria. Implicit treatment of solvation, such as using the generalized Born (GB) class of models arguably provides an optimal balance between computational efficiency and physical accuracy. Yet, GB models are frequently plagued by a tendency to generate overly compact structures. The physical origins of this drawback are relatively well understood, and the key to a balanced implicit solvent protein force field is careful optimization of physical parameters to achieve a sufficient level of cancellation of errors. The latter has been hampered by the difficulty of generating converged conformational ensembles of non-trivial model proteins using the popular replica exchange sampling technique. Here, we leverage improved sampling efficiency of a newly developed multi-scale enhanced sampling technique to re-optimize the generalized-Born with molecular volume (GBMV2) implicit solvent model with the CHARMM36 protein force field. Recursive optimization of key GBMV2 parameters (such as input radii) and protein torsion profiles (via the CMAP torsion cross terms) has led to a more balanced GBMV2 protein force field that recapitulates the structures and stabilities of both helical and β-hairpin model peptides. Importantly, this force field appears to be free of the over-compaction bias, and can generate structural ensembles of several intrinsically disordered proteins of various lengths that seem highly consistent with available experimental data. © 2017 Wiley Periodicals, Inc.