Dual-acting stapled peptides target both HIV-1 entry and assembly

Background: Previously, we reported the conversion of the 12-mer linear and cell-impermeable peptide CAI to a cell-penetrating peptide NYAD-1 by using an i,i + 4 hydrocarbon stapling technique and confirmed its binding to the C-terminal domain (CTD) of the HIV-1 capsid (CA) protein with an improved affinity (Kd ~ 1 μM) compared to CAI (Kd ~ 15 μM). NYAD-1 disrupts the formation of both immature- and mature-like virus particles in in vitro and cell-based assembly assays. In addition, it displays potent anti-HIV-1 activity in cell culture against a range of laboratory-adapted and primary HIV-1 isolates.<p></p> Results: In this report, we expanded the study to i,i + 7 hydrocarbon-stapled peptides to delineate their mechanism of action and antiviral activity. We identified three potent inhibitors, NYAD-36, -66 and -67, which showed strong binding to CA in NMR and isothermal titration calorimetry (ITC) studies and disrupted the formation of mature-like particles. They showed typical α-helical structures and penetrated cells; however, the cell penetration was not as efficient as observed with the i,i + 4 peptides. Unlike NYAD-1, the i,i + 7 peptides did not have any effect on virus release; however, they impaired Gag precursor processing. HIV-1 particles produced in the presence of these peptides displayed impaired infectivity. Consistent with an effect on virus entry, selection for viral resistance led to the emergence of two mutations in the gp120 subunit of the viral envelope (Env) glycoprotein, V120Q and A327P, located in the conserved region 1 (C1) and the base of the V3 loop, respectively.<p></p> Conclusion: The i,i + 7 stapled peptides derived from CAI unexpectedly target both CA and the V3 loop of gp120. This dual-targeted activity is dependent on their ability to penetrate cells as well as their net charge. This mechanistic revelation will be useful in further modifying these peptides as potent anti-HIV-1 agents.<p></p&gt

Three-Body Dynamics and Self-Powering of an Electrodynamic Tether in a Plasmasphere

The dynamics of an electrodynamic tether in a three-body gravitational environment are investigated. In the classical two-body scenario the extraction of power is at the expense of orbital kinetic energy. As a result of power extraction, an electrodynamic tether satellite system loses altitude and deorbits. This concept has been proposed and well investigated in the past, for example for orbital debris mitigation and spent stages reentry. On the other hand, in the three-body scenario an electrodynamic tether can be placed in an equilibrium position fixed with respect to the two primary bodies without deorbiting, and at the same time generate power for onboard use. The appearance of new equilibrium positions in the perturbed three-body problem allow this to happen as the electrical power is extracted at the expenses of the plasma corotating with the primary body. Fundamental differences between the classical twobody dynamics and the new phenomena appearing in the circular restricted three-body problem perturbed by the electrodynamic force of the electrodynamic tether are shown in the paper. An interesting application of an electrodynamic tether placed in the Jupiter plasma torus is then considered, in which the electrodynamic tether generates useful electrical power of about 1 kW with a 20-km-long electrodynamic tether from the environmental plasma without losing orbital energy

Dynamic stabilization of L2 periodic orbits using attitude-orbit coupling effects

Numerical explorations show how the known periodic solutions of the Hill problem are modified in the case of the attitude-orbit coupling that may occur for large satellite structures. We focus on the case in which the elongation is the dominant satellite?s characteristic and find that a rotating structure may remain with its largest dimension in a plane parallel to the plane of the primaries. In this case, the effect produced by the non-negligible physical dimension is dynamically equivalent to the perturbation produced by an oblate central body on a masspoint satellite. Based on this, it is demonstrated that the attitude-orbital coupling of a long enough body may change the dynamical characteristics of a periodic orbit about the collinear Lagrangian points

Quantifying signal changes in nano-wire based biosensors

In this work, we present a computational methodology for predicting the change in signal (conductance sensitivity) of a nano-BioFET sensor (a sensor based on a biomolecule binding another biomolecule attached to a nano-wire field effect transistor) upon binding its target molecule. The methodology is a combination of the screening model of surface charge sensors in liquids developed by Brandbyge and co-workers [Sørensen et al., Appl. Phys. Lett., 2007, 91, 102105], with the PROPKA method for predicting the pH-dependent charge of proteins and protein-ligand complexes, developed by Jensen and co-workers [Li et al., Proteins: Struct., Funct., Bioinf., 2005, 61, 704-721, Bas et al., Proteins: Struct., Funct., Bioinf., 2008, 73, 765-783]. The predicted change in conductance sensitivity based on this methodology is compared to previously published data on nano-BioFET sensors obtained by other groups. In addition, the conductance sensitivity dependence from various parameters is explored for a standard wire, representative of a typical experimental setup. In general, the experimental data can be reproduced with sufficient accuracy to help interpret them. The method has the potential for even more quantitative predictions when key experimental parameters (such as the charge carrier density of the nano-wire or receptor density on the device surface) can be determined (and reported) more accurately. © 2011 The Royal Society of Chemistry

Finite Element Models to Predict the Risk of Aseptic Loosening in Cementless Femoral Stems: A Literature Review

Aseptic loosening is the most common failure mode for total hip arthroplasty, and the design of the implant plays a significant role in influencing the longevity and stability of the implant. Finite Element (FE) models have been demonstrated to be powerful numerical tools that allow for generating information supporting the device’s safety and/or efficacy during pre-clinical assessment. Different authors have proposed FE studies aiming to simulate the long-term stability of the femoral stem; however, multiple improvements are still necessary for translating computational methodologies into clinical practice. This paper provides a comprehensive overview of the modelling procedures for predicting aseptic loosening risk, focusing on cementless femoral stems. The main modelling assumptions, including bone and implant geometry, materials, boundary conditions, and bone–implant interface contact, were summarised and presented. The limitations of various modelling assumptions and their impact on the simulation results were also discussed. The analysis suggests that more rigorous clinical validation for osseointegration models and failure criteria used to determine loosening of the implant should be clearly defined, and efforts should be made to identify the appropriate limit of tolerable conditions

POSITION PAPER : Credibility of in silico trial technologies - a theoretical framing

Different research communities have developed various approaches to assess the credibility of predictive models. Each approach usually works well for a specific type of model, and under some epistemic conditions that are normally satisfied within that specific research domain. Some regulatory agencies recently started to consider evidences of safety and efficacy on new medical products obtained using computer modelling and simulation (which is referred to as In Silico Trials); this has raised the attention in the computational medicine research community on the regulatory science aspects of this emerging discipline. But this poses a foundational problem: in the domain of biomedical research the use of computer modelling is relatively recent, without a widely accepted epistemic framing for problem of model credibility. Also, because of the inherent complexity of living organisms, biomedical modellers tend to use a variety of modelling methods, sometimes mixing them in the solution of a single problem. In such context merely adopting credibility approaches developed within other research community might not be appropriate. In this position paper we propose a theoretical framing for the problem of assessing the credibility of a predictive models for In Silico Trials, which accounts for the epistemic specificity of this research field and is general enough to be used for different type of models

A finite element model to simulate intraoperative fractures in cementless hip stem designs

Intraoperative femur fractures are a complication of hip arthroplasty, strongly related to the cementless stem design; this kind of fracture is not always recognised during surgery, and revision surgery may be necessary. The present study aimed to simulate intraoperative crack propagation during stem implantation using subject-specific quasi-static finite element models. Eleven subject-specific finite element femur models were built starting from CT data, and the implant pose and size of a non-commercial cementless stem were identified. The model boundary conditions were set with a compressive load from 1000 N to 10 000 N, to simulate the surgeon's hammering, and element deactivation was used to model the crack propagation. Two damage quantifiers were analysed to identify a threshold value that would allow us to assess if a fracture occurred. A methodology to assess the primary stability of the stem during insertion was also proposed, based on a push-out test. Crack propagation up to the surface was obtained in six patients; in two cases there was no crack generation, while in three patients the crack did not reach the external surface. This study demonstrates the possibility to simulate the propagation of the fracture intraoperatively during hip replacement surgery and generate quantitative information about the bone damage using a virtual cohort of simulated patients with anatomical and physiological variability

Experimental validation of a subject-specific finite element model of lumbar spine segment using digital image correlation

Pathologies such as cancer metastasis and osteoporosis strongly affect the mechanical properties of the vertebral bone and increase the risk of fragility fractures. The prediction of the fracture risk with a patient-specific model, directly generated from the diagnostic images of the patient, could help the clinician in the choice of the correct therapy to follow. But before such models can be used to support any clinical decision, their credibility must be demonstrated through verification, validation, and uncertainty quantification. In this study we describe a procedure for the generation of such patient-specific finite element models and present a first validation of the kinematics of the spine segment. Quantitative computed tomography images of a cadaveric lumbar spine segment presenting vertebral metastatic lesions were used to generate the model. The applied boundary conditions replicated a specific experimental test where the spine segment was loaded in compression-flexion. Model predictions in terms of vertebral surface displacements were compared against the full-field experimental displacements measured with Digital Image Correlation. A good agreement was obtained from the local comparison between experimental data and simulation results (R2 > 0.9 and RMSE% <8%). In conclusion, this work demonstrates the possibility to apply the developed modelling pipeline to predict the displacement field of human spine segment under physiological loading conditions, which is a first fundamental step in the credibility assessment of these clinical decision-support technology

Interaction of Polysialic Acid with CCL21 Regulates the Migratory Capacity of Human Dendritic Cells

Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs). Immature DCs (iDCs) are situated in the periphery where they capture pathogen. Subsequently, they migrate as mature DCs (mDCs) to draining lymph nodes to activate T cells. CCR7 and CCL21 contribute to the migratory capacity of the DC, but it is not completely understood what molecular requirements are involved. Here we demonstrate that monocyte-derived DCs dramatically change ST8Sia IV expression during maturation, leading to the generation of polysialic acid (polySia). PolySia expression is highly upregulated after 2 days Toll-like receptor-4 (TLR4) triggering. Surprisingly, only immunogenic and not tolerogenic mDCs upregulated polySia expression. Furthermore, we show that polySia expression on DCs is required for CCL21-directed migration, whereby polySia directly captures CCL21. Corresponding to polySia, the expression level of CCR7 is maximal two days after TLR4 triggering. In contrast, although TLR agonists other than LPS induce upregulation of CCR7, they achieve only a moderate polySia expression. In situ we could detect polySia-expressing APCs in the T cell zone of the lymph node and in the deep dermis. Together our results indicate that prolonged TLR4 engagement is required for the generation of polySia-expressing DCs that facilitate CCL21 capture and subsequent CCL21-directed migration

Direct peptide bioconjugation/PEGylation at tyrosine with linear and branched polymeric diazonium salts

Direct polymer conjugation at peptide tyrosine residues is described. In this study Tyr residues of both leucine enkephalin and salmon calcitonin (sCT) were targeted using appropriate diazonium salt-terminated linear monomethoxy poly(ethylene glycol)s (mPEGs) and poly(mPEG) methacrylate prepared by atom transfer radical polymerization. Judicious choice of the reaction conditions-pH, stoichiometry, and chemical structure of diazonium salt-led to a high degree of site-specificity in the conjugation reaction, even in the presence of competitive peptide amino acid targets such as histidine, lysines, and N-terminal amine. In vitro studies showed that conjugation of mPEG 2000 to sCT did not affect the peptide's ability to increase intracellular cAMP induced in T47D human breast cancer cells bearing sCT receptors. Preliminary in vivo investigation showed preserved ability to reduce [Ca 2+] plasma levels by mPEG 2000-sCT conjugate in rat animal models. © 2012 American Chemical Society