Modeling the self-assembly of functionalized fullerenes on solid surfaces using Monte Carlo simulations

Since their discovery 25 years ago, carbon fullerenes have been widely studied for their unique physicochemical properties and for applications including organic electronics and photovoltaics. For these applications it is highly desirable for crystalline fullerene thin films to spontaneously self-assemble on surfaces. Accordingly, many studies have functionalized fullerenes with the aim of tailoring their intermolecular interactions and controlling interactions with the solid substrate. The success of these rational design approaches hinges on the subtle interplay of intermolecular forces and molecule-substrate interactions. Molecular modeling is well-suited to studying these interactions by directly simulating self-assembly. In this work, we consider three different fullerene functionalization approaches and for each approach we carry out Monte Carlo simulations of the self-assembly process. In all cases, we use a coarse-grained molecular representation that preserves the dominant physical interactions between molecules and maximizes computational efficiency. The first approach we consider is the traditional gold-thiolate SAM (self-assembled monolayer) strategy which tethers molecules to a gold substrate via covalent sulfur-gold bonds. For this we study an asymmetric fullerene thiolate bridged by a phenyl group. Clusters of 40 molecules are simulated on the Au(111) substrate at different temperatures and surface coverage densities. Fullerenes and S atoms are found to compete for Au(111) surface sites, and this competition prevents self-assembly of highly ordered monolayers. Next, we investigate self-assembled monolayers formed by fullerenes with hydrogen-bonding carboxylic acid substituents. We consider five molecules with different dimensions and symmetries. Monte Carlo cooling simulations are used to find the most stable solid structures of clusters adsorbed to Au(111). The results show cases where fullerene-Au(111) attraction, fullerene close-packing, and hydrogen-bonding interactions can cooperate to guide self-assembly or compete to hinder it. Finally, we consider three bis-fullerene molecules, each with a different bridging group covalently joining two fullerenes. To effectively study the competing standing-up and lying-down morphologies, we use Monte Carlo simulations in conjunction with replica exchange and force field biasing methods. For clusters adsorbed to smooth model surfaces, we determine free energy landscapes and demonstrate their utility for rationalizing and predicting self-assembly

Examining the Relationship Between Training Environment and Muscle Dysmorphia Symptoms

Muscle Dysmorphia (MD) is characterized by preoccupation with muscularity. Although there is a growing body of research concerning MD, there is a lack of research concerning the potential role exercise training environment has on the clinical features of MD. The purpose of this study was to compare MD symptomology in traditional strength-trained (TRAD) individuals to individuals training in communal high intensity functional training environments (HIFT). Participants were recruited from both types of facilities. Participants (N=376) completed online (Qualtrics) demographics survey and Muscle Dysmorphia Inventory (MDI). One-way ANOVA compared the effect of training environment on MDI scores among HIFT, TRAD, both HIFT and TRAD (BOTH), home gym (HOME), and “OTHER.” Training environment significantly affected MDI for the 5 environments [F (4, 345) = 3.765, p = .005, d = 0.737]. Mean score for TRAD (M = 111.73, SD = 20. 39, [107.78, 115.68]) was significantly higher than HIFT (M = 102.20, SD = 19.59, [99.17, 105.23]). MDI for BOTH (M= 107.06, SD = 18.01, [100.77, 113.34]), HOME (M = 108.89, SD = 22.80, [99.86, 117.90]), and OTHER condition (M = 108.19, SD = 22.43, [97.97, 118.40]) did not significantly differ from HIFT or TRAD. Results suggest training environment is correlated with levels of MD symptomology. Specifically, males and females with higher levels of MD symptoms prefer to train in a traditional training environment, which is potentially more conducive to facilitating and perpetuating MD symptomology. The results of this study provide insight into the social physique anxiety associated with MD, as participants with higher levels of MD symptoms do not prefer to train in a HIFT environment where training occurs communally and other gym members provide extrinsic motivation. Additionally, the results of the present study further our understanding into the psychopathology of MD in that the motivating factors related to aesthetics (high level of body focus) associated with a TRAD environment take precedence over the motivating factors relating to selfimprovement and the desire to increase functional fitness that is associated with a HIFT environment. Results may provide knowledge for creating optimal treatment programs for individuals with clinical MD

Triple-helical collagen hydrogels via covalent aromatic functionalization with 1,3 phenylenediacetic acid

Chemical crosslinking of collagen is a general strategy to reproduce macroscale tissue properties in physiological environment. However, simultaneous control of protein conformation, material properties and biofunctionality is highly challenging with current synthetic strategies. Consequently, the potentially-diverse clinical applications of collagen-based biomaterials cannot be fully realised. In order to establish defined biomacromolecular systems for mineralised tissue applications, type I collagen was functionalised with 1,3-phenylenediacetic acid (Ph) and investigated at the molecular, macroscopic and functional levels. Preserved triple helix conformation was observed in obtained covalent networks via ATR-FTIR (AIII/A1450 [similar] 1) and WAXS, while network crosslinking degree (C: 87–99 mol%) could be adjusted based on specific reaction conditions. Decreased swelling ratio (SR: 823–1285 wt%) and increased thermo-mechanical (Td: 80–88 °C; E: 28–35 kPa; σmax: 6–8 kPa; εb: 53–58%) properties were observed compared to state-of-the-art carbodiimide (EDC)-crosslinked collagen controls, likely related to the intermolecular covalent incorporation of the aromatic segment. Ph-crosslinked hydrogels displayed nearly intact material integrity and only a slight mass decrease (MR: 5–11 wt%) following 1 week incubation in either PBS or simulated body fluid (SBF), in contrast to EDC-crosslinked collagen (MR: 33–58 wt%). Furthermore, FTIR, SEM and EDS revealed deposition of a calcium–phosphate phase on SBF-retrieved samples, whereby an increased calcium phosphate ratio (Ca/P: 0.84–1.41) was observed in hydrogels with higher Ph content. 72 hours material extracts were well tolerated by L929 mouse fibroblasts, whereby cell confluence and metabolic activity (MTS assay) were comparable to those of cells cultured in cell culture medium (positive control). In light of their controlled structure–function properties, these biocompatible collagen hydrogels represent attractive material systems for potential mineralised tissue applications

Photo-active collagen systems with controlled triple helix architecture

The design of photo-active collagen systems is presented as a basis for establishing biomimetic materials with varied network architecture and programmable macroscopic properties. Following in-house isolation of type I collagen, reaction with vinyl-bearing compounds of varied backbone rigidity, i.e. 4-vinylbenzyl chloride (4VBC) and glycidyl methacrylate (GMA), was carried out. TNBS colorimetric assay, 1H-NMR and ATR-FTIR confirmed covalent and tunable functionalization of collagen lysines. Depending on the type and extent of functionalization, controlled stability and thermal denaturation of triple helices were observed via circular dichroism (CD), whereby the hydrogen-bonding capability of introduced moieties was shown to play a major role. Full gel formation was observed following photo-activation of functionalized collagen solutions. The presence of a covalent network only slightly affected collagen triple helix conformation (as observed by WAXS and ATR-FTIR), confirming the structural organization of functionalized collagen precursors. Photo-activated hydrogels demonstrated an increased denaturation temperature (DSC) with respect to native collagen, suggesting that the formation of the covalent network successfully stabilized collagen triple helices. Moreover, biocompatibility and mechanical competence of obtained hydrogels were successfully demonstrated under physiologically-relevant conditions. These results demonstrate that this novel synthetic approach enabled the formation of biocompatible collagen systems with defined network architecture and programmable macroscopic properties, which can only partially be obtained with current synthetic methods

Low-Density Lipoprotein Has an Enormous Capacity To Bind (E)-4-Hydroxynon-2-enal (HNE): Detection and Characterization of Lysyl and Histidyl Adducts Containing Multiple Molecules of HNE

(E)-4-Hydroxynon-2-enal (HNE), an electrophilic bifunctional cytotoxic lipid peroxidation product, forms covalent adducts with nucleophilic side chains of amino acid residues. HNE-derived adducts have been implicated in many pathophysiological processes including atherosclerosis, diabetes, and Alzheimer’s disease. Tritium- and deuterium-labeled HNE (d4-HNE) were used orthogonally to study adduction with proteins and individual nucleophilic groups of histidyl, lysyl, and cysteine residues. Using tritium-labeled HNE, we detected the binding of 486 molecules of HNE per low-density lipoprotein (LDL) particle, significantly more than the total number of all reactive nucleophiles in the LDL particle. This suggests the formation of adducts that incorporate multiple molecules of HNE with some nucleophilic amino acid side chains. We also found that the reaction of a 1:1 mixture of d4-HNE and d0-HNE with N-acetylhistidine, N-acetyl-Gly-Lys-OMe, or N-acetyl cysteine generates 1:1, 2:1, and 3:1 adducts, which exhibit unique mass spectral signatures that aid in structural characterization. A domino-like reaction of initial 1:1 HNE Michael adducts of histidyl or lysyl nucleophiles with multiple additional HNE molecules forms 2:1 and 3:1 adducts that were structurally characterized by tandem mass spectrometry

A Streptomyces lividans SipY defficient strain as a host for protein production : standardization of operational alternatives for model proteins

Background: Extracellular protein production by Gram-positive bacteria, such as Streptomyces, may be complementary to current established protein production processes. The performance of a Streptomyces lividans mutant strain, deficient in the major signal peptidase (SipY) is investigated for the production of proteins secreted via the secondary Tat pathway. - Results: The SipY deficient strain has shown advantages over the wild type strain, in terms of extracellular productivity, specific activity and rheological behaviour. Two operational modes, batch and fed-batch, have been studied using mannitol as carbon source. The results showed that two successive mannitol additions in fed-batch mode led to improved secretory protein production using Streptomyces agarase as a model protein. This production process was also explored for the Tat secretory protein S. lividans laccase. The predicted sequence for the pre-laccase coding sequence has been cloned into the mutant strain under the control of the agarase promoter. Batch and fed-batch laccase production, using either mannitol or glucose as carbon source, have been developed and quantified. - Conclusions: The usefulness of a Streptomyces lividans SipY deficient strain as protein producer has been demonstrated. A proposed operating mode with substrate additions has been employed for the optimisation of Tat proteins production, although some adjustments might be necessary depending on the secretory protein

Fabrication and characterisation of electrospun silk fibroin/gelatin scaffolds crosslinked with glutaraldehyde vapour

Bombyx mori silk fibroin (SF) /gelatin nanofibre mats with different blend ratios of 100/0, 90/10 and 70/30 were prepared by electrospinning and crosslinked with glutaraldehyde (GTA) vapour at room temperature. GTA was shown to induce the conformational transition of SFs from random coils to β-sheets along with increasing nanofibre diameters with the addition of gelatin into SFs. It was found that by increasing the gelatin content, crosslinking degree was enhanced from 34% for pure SF nanofibre mats to 43% for SF/gelatin counterparts at the blend ratio of 70/30, which directly affected mechanical properties, porosity, and water uptake capacity (WUC) of prepared nanofibre mats. The addition of 10 and 30 wt% gelatin into SFs improved tensile strengths of SF/gelatin nanofibre mats by 10 and 27% along with significant increases in Young’s modulus by 1.1 and 1.3 times, respectively, as opposed to plain SF counterparts. However, both porosity and WUC were found to decrease from 62 and 405% for pristine SF nanofibre mats to 47% and 232% for SF/gelatin counterparts at the blend ratio of 70/30 accordingly. To further evaluate the combined effect of GTA crosslinking and gelatin content on biological response of SF/gelatin scaffolds, the proliferation assay using 3T3 mouse fibroblast was conducted. In comparison with pure SFs, cell proliferation rate was lower for SF/gelatin constructs, which declined when the gelatin content increased. These results indicated that the adverse effect of GTA crosslinking on cell response may be ascribed to imposed changes in morphology and physiochemical properties of SF/gelatin nanofibre mats. Although crosslinking could be used to improve mechanical properties of nanofibre mats, it reduced their capacity to support the cell activity. GTA optimisation is required to further modulate the physico-chemical properties of SF/gelatin nanofibre mats in order to obtain stable materials with favourable bioactive properties and promote cellular responses for tissue engineering applications