Genetic Bias,Diversity Indices,Physiochemical Properties and CDR3 Motifs Divide Auto-Reactive from Allo-Reactive T-Cell Repertoires

The distinct properties of allo-reactive T-cell repertoires are not well understood. To investigate whether auto-reactive and allo-reactive T-cell repertoires encoded distinct properties, we used dextramer enumeration, enrichment, single-cell T-cell receptor (TCR) sequencing and multiparameter analysis. We found auto-reactive and allo-reactive T-cells differed in mean ex vivo frequency which was antigen dependent. Allo-reactive T-cells showed clear differences in TCR architecture, with enriched usage of specific T-cell receptor variable (TRBJ) genes and broader use of T-cell receptor variable joining (TRBJ) genes. Auto-reactive T-cell repertoires exhibited complementary determining regions three (CDR3) lengths using a Gaussian distribution whereas allo-reactive T-cell repertoires exhibited distorted patterns in CDR3 length. CDR3 loops from allo-reactive T-cells showed distinct physical-chemical properties, tending to encode loops that were more acidic in charge. Allo-reactive T-cell repertoires differed in diversity metrics, tending to show increased overall diversity and increased homogeneity between repertoires. Motif analysis of CDR3 loops showed allo-reactive T-cell repertoires differed in motif preference which included broader motif use. Collectively, these data conclude that allo-reactive T-cell repertoires are indeed different to auto-reactive repertoires and provide tangible metrics for further investigations and validation. Given that the antigens studied here are overexpressed on multiple cancers and that allo-reactive TCRs often show increased ligand affinity, this new TCR bank also has translational potential for adoptive cell therapy, soluble TCR-based therapy and rational TCR design.     

Alteration of 3D Matrix Stiffness Regulates Viscoelasticity of Human Mesenchymal Stem Cells

Human mesenchymal stem cells (hMSCs) possess potential of bone formation and were proposed as ideal material against osteoporosis. Although interrogation of directing effect on lineage specification by physical cues has been proposed, how mechanical stimulation impacts intracellular viscoelasticity during osteogenesis remained enigmatic. Cyto-friendly 3D matrix was prepared with polyacrylamide and conjugated fibronectin. The hMSCs were injected with fluorescent beads and chemically-induced toward osteogenesis. The mechanical properties were assessed using video particle tracking microrheology. Inverted epifluorescence microscope was exploited to capture the Brownian trajectory of hMSCs. Mean square displacement was calculated and transformed into intracellular viscoelasticity. Two different stiffness of microspheres (12 kPa, 1 kPa) were established. A total of 45 cells were assessed. hMSCs possessed equivalent mechanical traits initially in the first week, while cells cultured in rigid matrix displayed significant elevation over elastic (G′) and viscous moduli (G″) on day 7 (p < 0.01) and 14 (p < 0.01). However, after two weeks, soft niches no longer stiffened hMSCs, whereas the effect by rigid substrates was consistently during the entire differentiation course. Stiffness of matrix impacted the viscoelasticity of hMSCs. Detailed recognition of how microenvironment impacts mechanical properties and differentiation of hMSCs will facilitate the advancement of tissue engineering and regenerative medicine.     

Experimental and numerical study of stratified viscous oil–water flow

Stratified two-phase flows of oil and water are important to the energy industry, and models capable of predicting this type of flow are primordial. Many studies focus on fluids with low viscosity, but a high viscosity oil in the mixture significantly changes its behavior. We gathered experimental data of pressure drop, volumetric fractions, and flow-pattern data of a stratified liquid–liquid flow with high viscosity ratio. In addition, a wire-mesh sensor provided tomographic views of the flow. The data were compared with computational fluid dynamics (CFD) models using OpenFOAM and a one-dimensional model. CFD simulations used an interface capturing method, and turbulence damping was introduced to avoid high eddy viscosity at the interface region. Reynolds Average Navier–Stokes and large eddy simulations were used to account for turbulence, and they showed significant differences. The comparisons showed good overall results for pressure drop, volumetric fractions, and phase distributions between CFD and experiments.     

The effect of stereoregularity on the thermal behavior of poly(methacrylic acid)s

The thermal degradation of a series of poly(methacrylic acid)s (PMADs) with different tacticities has been investigated under inert atmosphere. All the samples showed two main degradation stages. The first decomposition process of isotactic PMAD consisted of the formation of anhydride groups, almost quantitatively, by dehydration. In the case of PMADs with lower isotactic content, the elimination of water between neighboring carboxylic groups is less favored and the anhydridization shifts at higher temperatures where other degradation reactions may take place at the same time. The temperatures of maximum volatilization rates at the second step of decomposition increased in the order isotactic > atactic > syndiotactic. Received: 22 April 1997/Accepted: 13 May 1997     

Structure‐Based Design of a Eukaryote‐Selective Antiprotozoal Fluorinated Aminoglycoside

Aminoglycosides (AG) are antibiotics that lower the accuracy of protein synthesis by targeting a highly conserved RNA helix of the ribosomal A‐site. The discovery of AGs that selectively target the eukaryotic ribosome, but lack activity in prokaryotes, are promising as antiprotozoals for the treatment of neglected tropical diseases, and as therapies to read‐through point‐mutation genetic diseases. However, a single nucleobase change A1408G in the eukaryotic A‐site leads to negligible affinity for most AGs. Herein we report the synthesis of 6′‐fluorosisomicin, the first 6′‐fluorinated aminoglycoside, which specifically interacts with the protozoal cytoplasmic rRNA A‐site, but not the bacterial A‐site, as evidenced by X‐ray co‐crystal structures. The respective dispositions of 6′‐fluorosisomicin within the bacterial and protozoal A‐sites reveal that the fluorine atom acts only as a hydrogen‐bond acceptor to favorably interact with G1408 of the protozoal A‐site. Unlike aminoglycosides containing a 6′‐ammonium group, 6′‐fluorosisomicin cannot participate in the hydrogen‐bonding pattern that characterizes stable pseudo‐base‐pairs with A1408 of the bacterial A‐sites. Based on these structural observations it may be possible to shift the biological activity of aminoglycosides to act preferentially as antiprotozoal agents. These findings expand the repertoire of small molecules targeting the eukaryotic ribosome and demonstrate the usefulness of fluorine as a design element.     

Can the performance of graphene nanosheets for lithium storage in Li-ion batteries be predicted?

Graphene nanosheets (GNS) were prepared from graphitic oxide (GO) in two different ways: (a) thermal exfoliation at different temperatures; and (b) wet chemistry, using aqueous N(2)H(4) and KBH(4) as reducing agents. Irrespective of the synthetic method used, the materials exhibited a high irreversible capacity and strong polarization in their charge curves, when used in a Li-ion battery. The GNS synthesized with N(2)H(4) exhibited the best performance. Thus, at 149 mA g(-1) the average specific capacity delivered was ca. 600 mA h g(-1) after 100 cycles. On the other hand, the worst performance, irrespective of rate, was that of GNS synthesized with KBH(4) and the thermal GNS obtained at 800 °C. The physical and chemical analyses allowed various parameters to be derived for correlation with the electrochemical properties. Unfortunately, no clear-cut correlation was apparent. A comparison with reported data revealed that no correlation appears to exist with physical and chemical properties that allows a simple strategy for tailoring an effective graphene anode to be designed.     

A comparative study of two polyelectrolyte complexes

This study reports on the synthesis and characterization of interpenetrating polymeric network (IPN) and polyelectrolyte complexes (PEC) produced by ion-complex formation between natural and synthetic polymer bearing opposed charge. PEC of sodium alginate (NaAlg) and poly(acryloxyethyl-trimethylammonium chloride-co-2-hydroxyethyl methacrylate) (poly(Q-co-H)) were formed by addition of 2.0 wt % sodium alginate solution to 1.5 wt % poly(Q-co-H) solution, and by adding the poly(Q-co-H) solution to the NaAlg solution under magnetic agitation. The IPN were prepared using the sequential method with three different compositions, in which Q and H monomers were mixed in the 95/5 proportions in presence of NaAlg. The mixture was polymerized at 60°C using potassium persulfate as initiator. The characterizations of scaffolds were investigated by Fourier-transform infrared spectroscopy and thermogravimetric analysis. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012