Generation of carrier peptides for the delivery of nucleic acid drugs in primary cells

Now that the human genome has been decoded, the demand for novel therapeutic concepts, such as gene and stem cell therapy, is higher than ever before. Although new and better pharmaceutical agents are available, their efficient delivery to the intracellular site of action is still a serious challenge. A possible solution to this problem is the use of cell-penetrating peptides as delivery vectors, including derivatives of human calcitonin (hCT). The aim of this study was to synthesise novel branched hCT-derived peptides for the noncovalent delivery of nucleic acids. The uptake of the resulting oligocationic peptides into various cell lines as well as primary cells was monitored by fluorescence microscopy. To determine the appropriate peptide-plasmid charge ratios for efficient cell transfection, electromobility shift assays were carried out. Finally, flow cytometric and fluorescence microscopic studies of gene expression highlighted two novel hCT-derived peptides as highly effective in the delivery of noncovalently complexed plasmid DNA. Thus, the absence of cytotoxicity paired with highly efficient cell internalisation and transfection rates, in primary cells as well, make both peptides powerful candidates as drug delivery vectors, especially for plasmid DNA, for both in vivo and ex vivo therapeutic applications.     

Silicon Substitution for Aluminum in Calcium Silicate Hydrates

²⁷Al MAS and multiquantum (MQ) MASNMR (magic-angle spinning nuclear magnetic resonance) spectroscopy were used to study the substitution of silicon by aluminum in calcium silicate hydrates (C-S-H), which are the main component of hydrated portland cement. Synthetic C-S-H samples were prepared, and their chemical stability was studied. Two-dimensional 3Q-MASNMR spectra revealed the chemical shift and quadrupolar parameters (delta_iso, nu_Q) that labeled aluminum sites in the C-S-H. Tetrahedral aluminum was observed in the bridging and nonbridging sites of the silicate chains.     

Relationship between structure and rheological,mechanical and thermal properties of polylactide/Cloisite 30B nanocomposites

Melt‐state and solid state mechanical properties and thermal stability of polylactide layered silicate nanocomposites elaborated by melt intercalation were studied as a function of clay content. Wide angle X‐ray scattering results, transmission electron microscopy observations, and rheological measurements indicated that the clay was finely distributed in the polylactide matrix. Contrary to nonlinear mechanical properties, thermal and linear mechanical properties were shown to increase with increasing clay fraction. The nanoindentation measurements confirm the significant increase of linear mechanical properties previously observed by tensile tests. The good correlation of linear mechanical properties at the macrometric and nanometric scales is explained by the high dispersion degree of the nanofiller in the biodegradable polymer matrix. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Calcium carbonate and ammonium polyphosphate flame retardant additives formulated to protect ethylene vinyl acetate copolymer against fire: Hydrated or carbonated calcium?

In this study, the effect of the chemical nature of different calcium (Ca)-based minerals as flame retardant additives in combination with ammonium polyphosphate (APP), in 1:1 proportions, on the flame retardancy behavior and performance of ethylene vinyl acetate copolymer was discussed. Combining APP with partly and completely hydrated calcium oxide led to superior flame-retardant function detected in mass loss calorimeter measurements with respect to the corresponding system containing carbonated calcium. This privileged character was attributed to the higher reactivity of hydrated Ca-based fillers toward APP in comparison with Ca carbonate, which induced the formation of an intumescent residue. The difference between reactivity potential of hydrated and dry Ca was demonstrated by the newly formed thermally stable species, and further evidenced by thermogravimetric analysis performed on APP/fillers blends. Moreover, the presence of more crystalline domains in the Ca/phosphorus-based compounds was evidenced by XRD analysis of the mass loss calorimeter test residues. The results of this work highlight the role of blend additive systems on the performance of flame retardancy of polymer materials.     

Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis,Characterization, and Evaluation

There is an increasing interest in cationic polymers as important constituents of non-viral gene delivery vectors. In the present study, we developed a versatile synthetic route for the production of covalent polymeric conjugates consisting of water-soluble depolymerized chitosan (dCS; M_W 6–9 kDa) and low molecular weight polyethylenimine (PEI; 2.5 kDa linear, 1.8 kDa branched). dCS-PEI derivatives were evaluated based on their physicochemical properties, including purity, covalent bonding, solubility in aqueous media, ability for DNA condensation, and colloidal stability of the resulting polyplexes. They were complexed with non-integrating DNA vectors coding for reporter genes by simple admixing and assessed in vitro using liver-derived HuH-7 cells for their transfection efficiency and cytotoxicity. Using a rational screening cascade, a lead compound was selected (dCS-Suc-LPEI-14) displaying the best balance of biocompatibility, cytotoxicity, and transfection efficiency. Scale-up and in vivo evaluation in wild-type mice allowed for a direct comparison with a commercially available non-viral delivery vector (in vivo-jetPEI). Hepatic expression of the reporter gene luciferase resulted in liver-specific bioluminescence, upon intrabiliary infusion of the chitosan-based polyplexes, which exceeded the signal of the in vivo jetPEI reference formulation by a factor of 10. We conclude that the novel chitosan-derivative dCS-Suc-LPEI-14 shows promise and potential as an efficient polymeric conjugate for non-viral in vivo gene therapy.     

Differences in seasonal distribution of wild and stocked juvenile lake trout by depth and temperature in Lake Champlain

Lake trout (Salvelinus namaycush) reared in hatcheries are exposed to an environment and feeding regime that is different from wild lake trout, and are stocked at substantially larger sizes with higher lipid reserves. In addition to differences in diet and growth, this early experience may alter habitat use compared to the wild cohort. We used seasonal data on the depth and temperature distribution of wild and stocked juvenile lake trout to test for differences in habitat use and inform sampling strategies to evaluate annual recruitment. Bottom trawling was conducted from 2015 to 2019 in the central basin of Lake Champlain every two to four weeks during the ice-free season. Differences in distribution of wild and stocked lake trout were most pronounced during thermal stratification, when wild juveniles were more abundant than stocked juveniles at shallower depths and warmer temperatures and stocked juveniles were more abundant at deeper depths and colder temperatures. Temperature preferences may be a consequence of different early rearing environments; wild lake trout are acclimated to lake temperatures and forage, whereas stocked fish entered the lake with high lipid content and little foraging experience. Unbiased assessment of the proportion of wild lake trout and growth and survival of the entire juvenile lake trout population using bottom trawl sampling should either take place in the pre- and post-stratification seasons when wild and stocked fish are at the same depths, or include the full range of depths and temperatures that wild and stocked fish occupy during the stratified period.     

Biofuel or excavation? - Life cycle assessment (LCA) of soil remediation options

The environmental consequences of soil remediation through biofuel or through dig-and-dump were compared using life cycle assessment (LCA). Willow (Salix viminalis) was actually grown in-situ on a discontinued oil depot, as a phytoremediation treatment. These data were used for the biofuel remediation, while excavation-and-refill data were estimated from experience. The biofuel remediation had great environmental advantages compared to the ex situ excavation remediation. With the ReCiPe impact assessment method, which included biodiversity, the net environmental effect was even positive, in spite of the fact that the wood harvest was not utilised for biofuel production, but left on the contaminated site. Impact from the Salix viminalis cultivation was mainly through land use for the short rotation coppice, and through journeys of control personnel. The latter may be reduced when familiarity with biofuel as a soil treatment method increases. The excavation-and-refill remediation was dominated by the landfill and the transport of contaminated soil and backfill.     

Impact of ultrafiltration and nanofiltration of an industrial fish protein hydrolysate on its bioactive properties

BACKGROUND: Numerous studies have demonstrated that in vitro controlled enzymatic hydrolysis of fish and shellfish proteins leads to bioactive peptides. Ultrafiltration (UF) and/or nanofiltration (NF) can be used to refine hydrolysates and also to fractionate them in order to obtain a peptide population enriched in selected sizes. This study was designed to highlight the impact of controlled UF and NF on the stability of biological activities of an industrial fish protein hydrolysate (FPH) and to understand whether fractionation could improve its content in bioactive peptides. RESULTS: The starting fish protein hydrolysate exhibited a balanced amino acid composition, a reproducible molecular weight (MW) profile, and a low sodium chloride content, allowing the study of its biological activity. Successive fractionation on UF and NF membranes allowed concentration of peptides of selected sizes, without, however, carrying out sharp separations, some MW classes being found in several fractions. Peptides containing Pro, Hyp, Asp and Glu were concentrated in the UF and NF retentates compared to the unfractionated hydrolysate and UF permeate, respectively. Gastrin/cholecystokinin‐like peptides were present in the starting FPH, UF and NF fractions, but fractionation did not increase their concentration. In contrast, quantification of calcitonin gene‐related peptide (CGRP)‐like peptides demonstrated an increase in CGRP‐like activities in the UF permeate, relative to the starting FPH. The starting hydrolysate also showed a potent antioxidant and radical scavenging activity, and a moderate angiotensin‐converting enzyme (ACE)‐1 inhibitory activity, which were not increased by UF and NF fractionation. CONCLUSION: Fractionation of an FPH using membrane separation, with a molecular weight cut‐off adapted to the peptide composition, may provide an effective means to concentrate CGRP‐like peptides and peptides enriched in selected amino acids. The peptide size distribution observed after UF and NF fractionation demonstrates that it is misleading to characterize the fractions obtained by membrane filtration according to the MW cut‐off of the membrane only, as is currently done in the literature. Copyright © 2010 Society of Chemical Industry     

Heat and Mass Transfer in Capillary-Pumped Liquid Films in Square Cross-Section Minichannels

This study addresses heat and mass transfer during the vaporization of a liquid in a heated square cross-section mini-channel. A theoretical model is developed in steady state using the radius of curvature as a variable. One-dimensional simulations have been performed. An analysis of this model reveals that heat and mass transfer is governed by two main groups of non-dimensional numbers (i.e., Reynolds × Boiling and Weber × Boiling² numbers). Maps of heat transfer performance are thus proposed according to these non-dimensional numbers. A reduced model is finally derived, allowing the main parameters to be expressed (such as the extended meniscus length) analytically.