Fabrication of gradient hydrogels using a thermophoretic approach in microfluidics

The extracellular matrix presents spatially varying physical cues that can influence cell behavior in many processes. Physical gradients within hydrogels that mimic the heterogenous mechanical microenvironment are useful to study the impact of these cues on cellular responses. Therefore, simple and reliable techniques to create such gradient hydrogels are highly desirable. This work demonstrates the fabrication of stiffness gradient Gellan gum (GG) hydrogels by applying a temperature gradient across a microchannel containing hydrogel precursor solution. Thermophoretic migration of components within the precursor solution generates a concentration gradient that mirrors the temperature gradient profile, which translates into mechanical gradients upon crosslinking. Using this technique, GG hydrogels with stiffness gradients ranging from 20 to 90 kPa over 600 µm are created, covering the elastic moduli typical of moderately hard to hard tissues. MC3T3 osteoblast cells are then cultured on these gradient substrates, which exhibit preferential migration and enhanced osteogenic potential toward the stiffest region on the gradient. Overall, the thermophoretic approach provides a non-toxic and effective method to create hydrogels with defined mechanical gradients at the micron scale suitable for in vitro biological studies and potentially tissue engineering applications

Fabrication of gradient hydrogels using a thermophoretic approach in microfluidics

The extracellular matrix presents spatially varying physical cues that can influence cell behavior in many processes. Physical gradients within hydrogels that mimic the heterogenous mechanical microenvironment are useful to study the impact of these cues on cellular responses. Therefore, simple and reliable techniques to create such gradient hydrogels are highly desirable. This work demonstrates the fabrication of stiffness gradient Gellan gum (GG) hydrogels by applying a temperature gradient across a microchannel containing hydrogel precursor solution. Thermophoretic migration of components within the precursor solution generates a concentration gradient that mirrors the temperature gradient profile, which translates into mechanical gradients upon crosslinking. Using this technique, GG hydrogels with stiffness gradients ranging from 20 to 90 kPa over 600 µm are created, covering the elastic moduli typical of moderately hard to hard tissues. MC3T3 osteoblast cells are then cultured on these gradient substrates, which exhibit preferential migration and enhanced osteogenic potential toward the stiffest region on the gradient. Overall, the thermophoretic approach provides a non-toxic and effective method to create hydrogels with defined mechanical gradients at the micron scale suitable for in vitro biological studies and potentially tissue engineering applications

Muon capture by 3He nuclei followed by proton and deuteron production

The paper describes an experiment aimed at studying muon capture by ${}^{3}\mathrm{He}$ nuclei in pure ${}^{3}\mathrm{He}$ and $\mathrm{D}_2 + {}^{3}\mathrm{He}$ mixtures at various densities. Energy distributions of protons and deuterons produced via $\mu^-+{}^{3}\mathrm{He}\to p+n+n + \nu_{\mu }$ and $\mu^-+{}^{3} \mathrm{He} \to d+n + \nu_{\mu}$ are measured for the energy intervals $10 - 49$ MeV and $13 - 31$ MeV, respectively. Muon capture rates, $\lambda_\mathrm{cap}^p (\Delta E_p)$ and $\lambda_\mathrm{cap}^d (\Delta E_d)$ are obtained using two different analysis methods. The least--squares methods gives $\lambda_\mathrm{cap}^p = (36.7\pm 1.2) {s}^{- 1}$, $\lambda_\mathrm{cap}^d = (21.3 \pm 1.6) {s}^{- 1}$. The Bayes theorem gives $\lambda_\mathrm{cap}^p = (36.8 \pm 0.8) {s}^{- 1}$, $\lambda_\mathrm{cap}^d = (21.9 \pm 0.6) {s}^{- 1}$. The experimental differential capture rates, $d\lambda_\mathrm{cap}^p (E_p) / dE_p$ and $d\lambda_\mathrm{cap}^d (E_d) / dE_d$, are compared with theoretical calculations performed using the plane--wave impulse approximation (PWIA) with the realistic NN interaction Bonn B potential. Extrapolation to the full energy range yields total proton and deuteron capture rates in good agreement with former results.Comment: 17 pages, 13 figures, accepted for publication in PR

A prospective, multicentre, observational cohort study of analgesia and outcome after pneumonectomy

Background Meta-analysis and systematic reviews of epidural compared with paravertebral blockade analgesia techniques for thoracotomy conclude that although the analgesia is comparable, paravertebral blockade has a better short-term side-effect profile. However, reduction in major complications including mortality has not been proven. Methods The UK pneumonectomy study was a prospective observational cohort study in which all UK thoracic surgical centres were invited to participate. Data presented here relate to the mode of analgesia and outcome. Data were analysed for 312 patients having pneumonectomy at 24 UK thoracic surgical centres in 2005. The primary endpoint was a major complication. Results The most common type of analgesia used was epidural (61.1%) followed by paravertebral infusion (31%). Epidural catheter use was associated with major complications (odds ratio 2.2, 95% confidence interval 1.1–3.8; P=0.02) by stepwise logistic regression analysis. Conclusions An increased incidence of clinically important major post-pneumonectomy complications was associated with thoracic epidural compared with paravertebral blockade analgesia. However, this study is unable to provide robust evidence to change clinical practice for a better clinical outcome. A large multicentre randomized controlled trial is now needed to compare the efficacy, complications, and cost-effectiveness of epidural and paravertebral blockade analgesia after major lung resection with the primary outcome of clinically important major morbidity

Calcium Silicate-Based Biocompatible Light-Curable Dental Material for Dental Pulpal Complex

Dental caries causes tooth defects and clinical treatment is essential. To prevent further damage and protect healthy teeth, appropriate dental material is a need. However, the biocompatibility of dental material is needed to secure the oral environment. For this purpose, biocompatible materials were investigated for incorporated with dental capping material. Among them, nanomaterials are applied to dental materials to enhance their chemical, mechanical, and biological properties. This research aimed to study the physicochemical and mechanical properties and biocompatibility of a recently introduced light-curable mineral trioxide aggregate (MTA)-like material without bisphenol A-glycidyl methacrylate (Bis-GMA). To overcome the compromised mechanical properties in the absence of Bis-GMA, silica nanoparticles were synthesized and blended with a dental polymer for the formation of a nano-network. This material was compared with a conventional light-curable MTA-like material that contains Bis-GMA. Investigation of the physiochemical properties followed ISO 4049. Hydroxyl and calcium ion release from the materials was measured over 21 days. The Vickers hardness test and three-point flexural strength test were used to assess the mechanical properties. Specimens were immersed in solutions that mimicked human body plasma for seven days, and surface characteristics were analyzed. Biological properties were assessed by cytotoxicity and biomineralization tests. There was no significant difference between the tested materials with respect to overall physicochemical properties and released calcium ions. The newly produced material released more calcium ions on the third day, but 14 days later, the other material containing Bis-GMA released higher levels of calcium ions. The microhardness was reduced in a low pH environment, and differences between the specimens were observed. The flexural strength of the newly developed material was significantly higher, and different surface morphologies were detected. The recently produced extract showed higher cell viability at an extract concentration of 100%, while mineralization was clear at the conventional concentration of 25%. No significant changes in the physical properties between Bis-GMA incorporate material and nanoparticle incorporate materials

Protein Condensate Atlas from predictive models of heteromolecular condensate composition

Biomolecular condensates help cells organise their content in space and time. Cells harbour a variety of condensate types with diverse composition and many are likely yet to be discovered. Here, we develop a methodology to predict the composition of biomolecular condensates. We first analyse available proteomics data of cellular condensates and find that the biophysical features that determine protein localisation into condensates differ from known drivers of homotypic phase separation processes, with charge mediated protein-RNA and hydrophobicity mediated protein-protein interactions playing a key role in the former process. We then develop a machine learning model that links protein sequence to its propensity to localise into heteromolecular condensates. We apply the model across the proteome and find many of the top-ranked targets outside the original training data to localise into condensates as confirmed by orthogonal immunohistochemical staining imaging. Finally, we segment the condensation-prone proteome into condensate types based on an overlap with biomolecular interaction profiles to generate a Protein Condensate Atlas. Several condensate clusters within the Atlas closely match the composition of experimentally characterised condensates or regions within them, suggesting that the Atlas can be valuable for identifying additional components within known condensate systems and discovering previously uncharacterised condensates

Gene transfer into hepatocytes using asialoglycoprotein receptor mediated endocytosis of DNA complexed with an artificial tetra-antennary galactose ligand

We have constructed an artificial ligand for the hepatocyte-specific asialoglycoprotein receptor for the purpose of generating a synthetic delivery system for DNA. This ligand has a tetra-antennary structure, containing four terminal galactose residues on a branched carrier peptide. The carbohydrate residues of this glycopeptide were introduced by reductive coupling of lactose to the alpha- and epsilon-amino groups of the two N-terminal lysines on the carrier peptide. The C-terminus of the peptide, containing a cysteine separated from the branched N-terminus by a 10 amino acid spacer sequence, was used for conjugation to 3-(2-pyridyldithio)propionate-modified polylysine via disulfide bond formation. Complexes containing plasmid DNA bound to these galactose-polylysine conjugates have been used for asialoglycoprotein receptor-mediated transfer of a luciferase gene into human (HepG2) and murine (BNL CL.2) hepatocyte cell lines. Gene transfer was strongly promoted when amphipathic peptides with pH-controlled membrane-disruption activity, derived from the N-terminal sequence of influenza virus hemagglutinin HA-2, were also present in these DNA complexes. Thus, we have essentially borrowed the small functional domains of two large proteins, asialoglycoprotein and hemagglutinin, and assembled them into a supramolecular complex to generate an efficient gene-transfer system

Nanobodies raised against monomeric alpha-synuclein inhibit fibril formation and destabilize toxic oligomeric species

BACKGROUND: The aggregation of the protein ɑ-synuclein (ɑS) underlies a range of increasingly common neurodegenerative disorders including Parkinson’s disease. One widely explored therapeutic strategy for these conditions is the use of antibodies to target aggregated ɑS, although a detailed molecular-level mechanism of the action of such species remains elusive. Here, we characterize ɑS aggregation in vitro in the presence of two ɑS-specific single-domain antibodies (nanobodies), NbSyn2 and NbSyn87, which bind to the highly accessible C-terminal region of ɑS. RESULTS: We show that both nanobodies inhibit the formation of ɑS fibrils. Furthermore, using single-molecule fluorescence techniques, we demonstrate that nanobody binding promotes a rapid conformational conversion from more stable oligomers to less stable oligomers of ɑS, leading to a dramatic reduction in oligomer-induced cellular toxicity. CONCLUSIONS: The results indicate a novel mechanism by which diseases associated with protein aggregation can be inhibited, and suggest that NbSyn2 and NbSyn87 could have significant therapeutic potential

'I just want to watch the match': a practitioner's reflective account of men's health themed match day events at an English Premier League football club

This study reflects on the effectiveness and delivery of a series of health themed match day events at an English Premier League Football Club which aimed to create awareness and motivate men to adopt recommended health behaviours. A range of marketing techniques and activities were adopted within a targeted space and time to increase men's exposure to health information. The first author adopted a practitioner-cum-researcher role and was immersed in the planning and delivery of the events utilising the principles of ethnography. Data were predominately collated through observations and personal reflections logged via autobiographical field notes. Data were analysed through abductive reasoning. In general, men were reluctant to engage in health-related behaviours on match days. However, subtle, non-invasive approaches were deemed successful. Positive outcomes and case studies from the latter techniques are presented and suggestions for effective strategies that will better engage men in health information and behaviours are made. © 2014 © 2014 Taylor & Francis

High--order connected moments expansion for the Rabi Hamiltonian

We analyze the convergence properties of the connected moments expansion (CMX) for the Rabi Hamiltonian. To this end we calculate the moments and connected moments of the Hamiltonian operator to a sufficiently large order. Our large--order results suggest that the CMX is not reliable for most practical purposes because the expansion exhibits considerable oscillations.Comment: 12 pages, 5 figures, 1 tabl