V(2)O(5) nanoparticles obtained from a synthetic bariandite-like vanadium oxide: Synthesis, characterization and electrochemical behavior in an ionic liquid

Highly stable and crystalline V(2)O(5) nanoparticles with an average diameter of 15 nm have been easily prepared by thermal treatment of a bariandite-like vanadium oxide, V(10)O(24)center dot 9H(2)O. Their characterization was carried out by powder X-ray diffractometry (XRD). Fourier transform infrared (FT-IR) and Raman spectroscopies, and transmission electron microscopy (TEM). The fibrous and nanostructured film obtained by electrophoretic deposition of the V(2)O(5) nanoparticles showed good electroactivity when submitted to cyclic voltammetry in an ionic liquid-based electrolyte. The use of this film for the preparation of a nanostructured electrode led to an improvement of about 50% in discharge capacity values when compared with similar electrodes obtained by casting of a V(2)O(5) xerogel. (C) 2009 Elsevier Inc. All rights reserved.

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.

A novel physiological property of snake bradykinin-potentiating peptides-Reversion of MK-801 inhibition of nicotinic acetylcholine receptors

The first naturally occurring angiotensin-converting enzyme (ACE) inhibitors described are pyroglutamyl proline-rich oligopeptides, found in the venom of the viper Bothrops jararaca, and named as bradykinin-potentiating peptides (BPPs). Biochemical and pharmacological properties of these peptides were essential for the development of Captopril, the first active site-directed inhibitor of ACE, currently used for the treatment of human hypertension. However, a number of data have suggested that the pharmacological activity of BPPs could not only be explained by their inhibitory action on enzymatic activity of somatic ACE. In fact, we showed recently that the strong and long-lasting anti-hypertensive effect of BPP-10c [

A convenient synthetic route for alkynylselenides from alkynyl bromides and diaryl diselenides employing copper(I)/imidazole as novel catalyst system

A mild, convenient, and effective strategy is developed for the synthesis of alkynyl selenides from alkynyl bromides and respective diselenides using Cul/imidazole as a novel catalyst system with Mg as additive. The Procedure affords the title compounds in moderate to good yield (51-89%). The main advantages of the protocol include the use of inexpensive copper catalyst, a novel Cu(I)/imidazole combination, and good yield of the products. (C) 2008 Elsevier Ltd. All rights reserved.

Epichlorohydrin Cross-Linking of Synthetic DNA Oligomers

Epichlorohydrin (ECH), an important chemical in the synthetic polymer industry, is a bifunctional alkylating agent with the potential to form DNA interstrand crosslinks. Occupational exposure to this suspect carcinogen leads to chromosomal aberrations, and ECH has been shown to undergo reaction with DNA in vivo and in vitro. We are using denaturing polyacrylamide gel electrophoresis to assess cross-linking of synthetic DNA oligomers by both ECH and the related compound, epibromohydrin (EBH). Both epihalohydrins produce a low-mobility band on denaturing gels consistent with an interstrand cross-link. Moreover, the efficiencies, sequence preferences, reaction kinetics, and pH dependence differ for the two compounds, suggesting different mechanisms of reaction. Understanding these alkylation reactions may help explain the role of the epihalohydrins in cancer development.

Purification and analysis of protein synthetic initiation factors from Volvox Carteri

Volvox carteri, a multi-celled green algae, can grow synchronously given a sixteen hour light period followed by an eight hour dark period, a cycle which is repeated for a 48 hour growth cycle total. Near the end of each light period, reproductive cells divide rapidly resulting in the differentiation of ceIls. When the dark period begins, this differentiation stops and the cells remain dormant with little protein synthesis or differentiation occurring. Immediately after the lights come back on, however, the cells again undergo rapid protein synthesis and complete their differentiation. Previous studies have concluded that Volvox carteri discontinue protein synthesis during the dark phase due to regulation at the translational level and not the transcriptional level. Therefore, the inhibition of protein synthesis does not lie in the transfer of the protein coding sequence from DNA to mRNA, but rather in the transfer of this information from the mRNA to the ribosomes. My research examined this translational regulation to determine the factor(s) causing the discontinuation of protein synthesis during the dark phase. Evidence from other research further suggests that the control of translation lies in the initiation step rather than the elongation step. Eukaryotic initiation factors aid in the binding of the ribosomal subunits to the mRNA to initiate protein synthesis. It is known that initiation factors can be modified by phosphorylation, regulating their activity. Therefore, my study focused upon isolating some of these initiation factors in order to determine whether or not such modifications are responsible for the inhibition of dark phase protein synthesis in Volvox carteri.

Cherry picking with synthetic controls

The synthetic control (SC) method has been recently proposed as an alternative method to estimate treatment e ects in comparative case studies. Abadie et al. [2010] and Abadie et al. [2015] argue that one of the advantages of the SC method is that it imposes a data-driven process to select the comparison units, providing more transparency and less discretionary power to the researcher. However, an important limitation of the SC method is that it does not provide clear guidance on the choice of predictor variables used to estimate the SC weights. We show that such lack of speci c guidances provides signi cant opportunities for the researcher to search for speci cations with statistically signi cant results, undermining one of the main advantages of the method. Considering six alternative speci cations commonly used in SC applications, we calculate in Monte Carlo simulations the probability of nding a statistically signi cant result at 5% in at least one speci cation. We nd that this probability can be as high as 13% (23% for a 10% signi cance test) when there are 12 pre-intervention periods and decay slowly with the number of pre-intervention periods. With 230 pre-intervention periods, this probability is still around 10% (18% for a 10% signi cance test). We show that the speci cation that uses the average pre-treatment outcome values to estimate the weights performed particularly bad in our simulations. However, the speci cation-searching problem remains relevant even when we do not consider this speci cation. We also show that this speci cation-searching problem is relevant in simulations with real datasets looking at placebo interventions in the Current Population Survey (CPS). In order to mitigate this problem, we propose a criterion to select among SC di erent speci cations based on the prediction error of each speci cations in placebo estimations

Functionalization of dendrimers for improved gene delivery to mesenchymal stem cell

Disease, injury, and age problems compromise human quality of life and continuously motivate the search for new and more efficacious therapeutic approaches. The field of Tissue Regeneration and Engineering has greatly evolved over the last years, mainly due to the combination of the important advances verified in Biomaterials Science and Engineering with those of Cell and Molecular Biology. In particular, a new and promising area arose – Nanomedicine – that takes advantage of the extremely small size and especial chemical and physical properties of Nanomaterials, offering powerful tools for health improvement. Research on Stem Cells, the self-renewing progenitors of body tissues, is also challenging to the medical and scientific communities, being expectable the appearance of new and exciting stem cell-based therapies in the next years. The control of cell behavior (namely, of cell proliferation and differentiation) is of key importance in devising strategies for Tissue Regeneration and Engineering. Cytokines, growth factors, transcription factors and other signaling molecules, most of them proteins, have been identified and found to regulate and support tissue development and regeneration. However, the application of these molecules in long-term regenerative processes requires their continuous presence at high concentrations as they usually present short half-lives at physiological conditions and may be rapidly cleared from the body. Alternatively, genes encoding such proteins can be introduced inside cells and be expressed using cell’s machinery, allowing an extended and more sustained production of the protein of interest (gene therapy). Genetic engineering of stem cells is particularly attractive because of their self-renewal capability and differentiation potential. For Tissue Regeneration and Engineering purposes, the patient’s own stem cells can be genetically engineered in vitro and, after, introduced in the body (with or without a scaffold) where they will not only modulate the behavior of native cells (stem cell-mediated gene therapy), but also directly participate in tissue repair. Cells can be genetically engineered using viral and non-viral systems. Viruses, as a result of millions of years of evolution, are very effective for the delivery of genes in several types of cells, including cells from primary sources. However, the risks associated with their use (like infection and immunogenic reactions) are driving the search for non-viral systems that will efficiently deliver genetic material into cells. Among them, chemical methods that are promising and being investigated use cationic molecules as carriers for DNA. In this case, gene delivery and gene expression level remain relatively low when primary cells are used. The main goal of this thesis was to develop and assess the in vitro potential of polyamidoamine (PAMAM) dendrimers based carriers to deliver genes to mesenchymal stem cells (MSCs). PAMAM dendrimers are monodispersive, hyperbranched and nanospherical molecules presenting unique characteristics that make them very attractive vehicles for both drug and gene delivery. Although they have been explored for gene delivery in a wide range of cell lines, the interaction and the usefulness of these molecules in the delivery of genes to MSCs remains a field to be explored. Adult MSCs were chosen for the studies due to their potential biomedical applications (they are considered multipotent cells) and because they present several advantages over embryonic stem cells, such as easy accessibility and the inexistence of ethical restrictions to their use. This thesis is divided in 5 interconnected chapters. Chapter I provides an overview of the current literature concerning the various non-viral systems investigated for gene delivery in MSCs. Attention is devoted to physical methods, as well as to chemical methods that make use of polymers (natural and synthetic), liposomes, and inorganic nanoparticles as gene delivery vectors. Also, it summarizes the current applications of genetically engineered mesenchymal stem cells using non-viral systems in regenerative medicine, with special focus on bone tissue regeneration. In Chapter II, the potential of native PAMAM dendrimers with amine termini to transfect MSCs is evaluated. The level of transfection achieved with the dendrimers is, in a first step, studied using a plasmid DNA (pDNA) encoding for the β-galactosidase reporter gene. The effect of dendrimer’s generation, cell passage number, and N:P ratio (where N= number of primary amines in the dendrimer; P= number of phosphate groups in the pDNA backbone) on the level of transfection is evaluated, being the values always very low. In a second step, a pDNA encoding for bone morphogenetic protein-2, a protein that is known for its role in MSCs proliferation and differentiation, is used. The BMP-2 content produced by transfected cells is evaluated by an ELISA assay and its effect on the osteogenic markers is analyzed through several classical assays including alkaline phosphatase activity (an early marker of osteogenesis), osteocalcin production, calcium deposition and mineralized nodules formation (late osteogenesis markers). Results show that a low transfection level is enough to induce in vitro osteogenic differentiation in MSCs. Next, from Chapter III to Chapter V, studies are shown where several strategies are adopted to change the interaction of PAMAM dendrimers with MSCs cell membrane and, as a consequence, to enhance the levels of gene delivery. In Chapter III, generations 5 and 6 of PAMAM dendrimers are surface functionalized with arginine-glycine-aspartic acid (RGD) containing peptides – experiments with dendrimers conjugated to 4, 8 and 16 RGD units were performed. The underlying concept is that by including the RGD integrin-binding motif in the design of the vectors and by forming RGD clusters, the level of transfection will increase as MSCs highly express integrins at their surface. Results show that cellular uptake of functionalized dendrimers and gene expression is enhanced in comparison with the native dendrimers. Furthermore, gene expression is dependent on both the electrostatic interaction established between the dendrimer moiety and the cell surface and the nanocluster RGD density. In Chapter IV, a new family of gene delivery vectors is synthesized consisting of a PAMAM dendrimer (generation 5) core randomly linked at the periphery to alkyl hydrophobic chains that vary in length and number. Herein, the idea is to take advantage of both the cationic nature of the dendrimer and the capacity of lipids to interact with biological membranes. These new vectors show a remarkable capacity for internalizing pDNA, being this effect positively correlated with the –CH2– content present in the hydrophobic corona. Gene expression is also greatly enhanced using the new vectors but, in this case, the higher efficiency is shown by the vectors containing the smallest hydrophobic chains. Finally, chapter V reports the synthesis, characterization and evaluation of novel gene delivery vectors based on PAMAM dendrimers (generation 5) conjugated to peptides with high affinity for MSCs membrane binding - for comparison, experiments are also done with a peptide with low affinity binding properties. These systems present low cytotoxicity and transfection efficiencies superior to those of native dendrimers and partially degraded dendrimers (Superfect®, a commercial product). Furthermore, with this biomimetic approach, the process of gene delivery is shown to be cell surface receptor-mediated. Overall, results show the potential of PAMAM dendrimers to be used, as such or modified, in Tissue Regeneration and Engineering. To our knowledge, this is the first time that PAMAM dendrimers are studied as gene delivery vehicles in this context and using, as target, a cell type with clinical relevancy. It is shown that the cationic nature of PAMAM dendrimers with amine termini can be synergistically combined with surface engineering approaches, which will ultimately result in suitable interactions with the cytoplasmic membrane and enhanced pDNA cellular entry and gene expression. Nevertheless, the quantity of pDNA detected inside cell nucleus is always very small when compared with the bigger amount reaching cytoplasm (accumulation of pDNA is evident in the perinuclear region), suggesting that the main barrier to transfection is the nuclear membrane. Future work can then be envisaged based on the versatility of these systems as biomedical molecular materials, such as the conjugation of PAMAM dendrimers to molecules able to bind nuclear membrane receptors and to promote nuclear translocation.

Effect of the synthetic coumarin, ethyl 2-oxo-2H-chromene-3-carboxylate, on activity of Crotalus durissus ruruima sPLA2 as well as on edema and platelet aggregation induced by this factor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Synthetic amides toxic to the leaf-cutting ant Atta sexdens rubropilosa L. and its symbiotic fungus

1 Nine synthetic amides similar to natural N-piperidine-3-(4,5-methylenedioxyphenyl)-2-(E)-propenainide and N-pyrrolidine-3-(4,5-methylenedyoxiphenyl)2-(E)-propenamide were synthesized and identified by their spectroscopic data.2 the toxicity of these synthetic amides to the Atta sexdens rubropilosa workers and the antifungal activity against Leticoagaricus gongylophorus, the symbiotic fungus of the leaf-cutting ants, were determined.3 Workers ants that were fed daily on an artificial diet to which these compounds were added had a higher mortality rate than the controls for N-pyrrolidine-3(3',4'-methylenedioxyphenyl)-2-(E)-propenamide and N-benzyl-3-(3',4'-methylenedioxyphenyl)-2-(E)-propenamide at a concentration of 100 mu g/mL.4 the completely inhibition (100%) of the fungal growth was observed with N-piperldine-3-(3',4'-methylenedioxyphenyl)-2-(E)-propenamide and N,N-diethyl-3-(3',4'-methylenedioxyphenyl)-2-(E)-propenamide at concentrations of 50 and 100 mu g/mL and N-pirrolidine-3-(3',4'-methylenedioxyphenyl)-2-(E)-propenamide at a concentration of 100 mu g/mL.5 the possibility of controlling these insects in the future using synthetic piperamides that can simultaneously target both organisms is discussed.

Mortality of Bemisia tabaci biotype B (sternorrhyncha: aleyrodidae) adults by aliphatic and aromatic synthetic sucrose esters

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Synthetic lethality between eIF5A and Ypt1 reveals a connection between translation and the secretory pathway in yeast

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Are light traps baited with kairomones effective in the capture of Lutzomyia longipalpis and Lutzomyia intermedia? An evaluation of synthetic human odor as an attractant for phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)