7 research outputs found
Microwave-assisted solid-phase peptide synthesis at 60 degrees C: alternative conditions with low enantiomerization
Several conditions have been used in the coupling reaction of stepwise SPPS at elevated temperature (SPPS-ET), but we have elected the following as our first choice: 2.5-fold molar excess of 0.04-0.08 M Boc or Fmoc-amino acid derivative, equimolar amount of DIC/HOBt (1:1)or TBTU/DIPEA(1:3), 25% DMSO/toluene, 60 degrees C, conventional heating. In this study, aimed to further examine enantiomerization under such condition and study the applicability of our protocols to microwave-SPPS, peptides containing L-Ser, L-His, L-Cys and/or L-Met were manually synthesized traditionally, at 60 degrees C using conventional heating and at 60 degrees C using microwave heating. Detailed assessment of all crude peptides (in their intact and/or fully hydrolyzed forms) revealed that, except for the microwave-assisted coupling of L-Cys, all other reactions occurred with low levels of amino acid enantiomerization (<2%). Therefore, herein we (i) provide new evidences that our protocols for SPPS at 60 degrees C using conventional heating are suitable for routine use, (ii) demonstrate their appropriateness for microwave-assisted SPPS by Boc and Fmoc chemistries, (iii) disclose advantages and limitations of the three synthetic approaches employed. Thus, this study complements our past research on SPPS-ET and suggests alternative conditions for microwave-assisted SPPS. Copyright (C) 2009 European Peptide Society and John Wiley & Sons, Ltd.FAPESPFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento CientÃfico e Tecnológico (CNPq)CNP
Surfactin application for a short period (10/20 s) increases the surface wettability of sound dentin
International audienceThe aim of this study was to evaluate the effect of spreading the lipopeptide surfactin, for short time (10/20 s), on dentin wettability. Study groups were surfactin: 2.8; 1.4; 0.7; 0.35; and 0.175 mg/mL and a control group that received no treatment. Dentin discs (4 mm height) were prepared and polished with 600-grit SiC paper. Contact angle determinations were carried out after microbrush spreading of surfactin on dentin specimens for, respectively, 10 and 20 s. Excess liquid was removed, and after 60 s, the specimens were analyzed in a goniometer using the sessile drop method to measure the contact angle. Results were analyzed by two-way ANOVA (concentration x time) and t student, with alpha = 0.05. Lower contact angles were obtained for surfactin (0.7 mg/mL) spread for 10 s. However, no statistical difference was observed for surfactin (2.8 mg/mL) applied during 20 s. Higher contact angles were observed for surfactin (0.7 mg/mL) spread for 20 s. In conclusion, dentin wettability is dependent on spreading time and surfactin concentration
Cell Penetrating Peptide (CPP)-Conjugated Desferrioxamine for Enhanced Neuroprotection: Synthesis and in Vitro Evaluation
Iron
overload causes progressive and sometimes irreversible damage
due to accelerated production of reactive oxygen species. Desferrioxamine
(DFO), a siderophore, has been used clinically to remove excess iron.
However, the applications of DFO are limited because of its inability
to access intracellular labile iron. Cell penetrating peptides (CPPs)
have become an efficient delivery vector for the enhanced internalization
of drugs into the cytosol. We describe, herein, an efficient method
for covalently conjugating DFO to the CPPs TATÂ(47–57) and Penetratin.
Both conjugates suppressed the redox activity of labile plasma iron
in buffered solutions and in iron-overloaded sera. Enhanced access
to intracellular labile iron compared to the parent siderophore was
achieved in HeLa and RBE4 (a model of blood-brain-barrier) cell lines.
Iron complexes of both conjugates also had better permeability in
both cell models. DFO antioxidant and iron binding properties were
preserved and its bioavailability was increased upon CPP conjugation,
which opens new therapeutic possibilities for neurodegenerative processes
associated with brain iron overload
Fluorimetric determination of intra- and extracellular free amino acids in the microalgae Tetraselmis gracilis (Prasinophyceae) using monolithic column in reversed phase mode
This paper describes the development and application of an RP HPLC method using a C(18) monolithic stationary phase for the separation and quantification of extra- and intracellular amino acids in a batch cultivation of the marine alga Tetraselmis gracilis. Fluorimetric detection was made after separation of the o-phthaldialdehyde 2-mercaptoethanol (OPA-2MCE) derivatives using a binary gradient elution. Separation of 19 amino acids was achieved with resolution >1.5 in about 39 min at a flow rate of 1.5 mL/min. RSD of analyses in seawater medium ranged from 0.36% for Orn (0.50 mu mol/L) to 12% for Ile (0.10 mu mol/L). The main constituents of the intracellular dissolved free amino acids (DFAAs) in the exponential growth phase were arginine (Arg), asparagine (Asn), alanine (Ala), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), glycine (Gly), glutamine (Gln), and leucine (Leu). The major amino acids excreted to the media were valine (Val), Ala, Ser, and Gly. The monolithic phase facilitates the analysis by shortening the separation time and saving solvents and instrumentation costs (indeed conventional HPLC instrumentation can be used, running at lower pressures than those ones used with packed particle columns).Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvintento Cientifico e Tecnologico (CNPq)Conselho Nacional de Desenvolvimento CientÃfico e Tecnológico (CNPq
5-Hydroxymethylfurfural and Furfural Base-Free Oxidation over AuPd Embedded Bimetallic Nanoparticles
International audienceThe heterogeneous catalytic partial oxidation of alcohols and aldehydes in the liquid phase usually needs the addition of a homogeneous base, which in turn makes the products’ recovery cumbersome, and can further induce undesired side reactions. In the present work, we propose the use of novel catalysts based on metallic Au, Pd and bimetallic AuPd nanoparticles embedded in a titanosilicate matrix. The as-prepared catalysts showed good efficiency in the base-free partial oxidation of furfural and 5-hydroxymethylfurfural. Au4Pd1@SiTi catalyst showed high selectivity (78%) to monoacids (namely, 5-formyl-2-furancarboxylic acid and 5-hydroxymethyl-2-furancarboxylic acid) at 50% 5-hydroxymethylfurfural (HMF) conversion. The selectivity even reached 83% in the case of furfural oxidation to furoic acid (at 50% furfural conversion). The performances of the catalysts strongly depended on the Au–Pd ratio, with an optimal value of 4:1. The pH of the solution was always below 3.5 and no leaching of metals was observed, confirming the stabilization of the metal nanoparticles within the titanosilicate host matrix