Nuclear magnetic resonance spectroscopic analysis of ethyl ester yield in the transesterification of vegetable oil: an accurate method for a truly quantitative analysis.

Biodiesel has recently gained importance as an alternative to fossil diesel. However, the development of more efficient biodiesel formation processes still depends on the use of fast and accurate analytical techniques to evaluate the conversion degree of the transesterification reaction. Nuclear magnetic resonance (NMR) spectroscopy has been used for this purpose, but some experimental details still need to be addressed. Therefore, in this communication, the experimental conditions for a truly quantitative NMR analysis of biodiesel formation are presented. The longitudinal relaxation time (T(1) ), which is the determining factor for quantitative analysis, was measured using an inversion-recovery method, and a maximum value of 2.35 s was obtained for a biodiesel sample. A linear determination coefficient of r(2) = 0.99 was obtained when a time delay between pulses longer than 5T(1) =15 s was used, whereas strong deviations were observed when using shorter delays.

A New Dermal Substitute Containing Polyvinyl Alcohol with Silver Nanoparticles and Collagen with Hyaluronic Acid: In Vitro and In Vivo Approaches.

The experimental use of poly (alcohol-vinyl) (PVA) as a skin curative is increasing widely. However, the use of this hydrogel is challenging due to its favorable properties for microbiota growth. The association with silver nanoparticles (AgNPs) as an antimicrobial agent turns the match for PVA as a dressing, as it focuses on creating a physical barrier to avoid wound dehydration. When associated with extracellular components, such as the collagen matrix, the device obtained can create the desired biological conditions to act as a skin substitute. This study aimed to analyze the anti-microbiological activity and the in vitro and in vivo responses of a bilaminar device of PVA containing AgNPs associated with a membrane of collagen-hyaluronic acid (col-HA). Additionally, mesenchymal stem cells were cultured in the device to evaluate in vitro responses and in vivo immunomodulatory and healing behavior. The device morphology revealed a porous pattern that favored water retention and in vitro cell adhesion. Controlled wounds in the dorsal back of rat skins revealed a striking skin remodeling with new epidermis fulfilling all previously injured areas after 14 and 28 days. No infections or significant inflammations were observed, despite increased angiogenesis, and no fibrosis-markers were identified as compared to controls. Although few antibacterial activities were obtained, the addition of AgNPs prevented fungal growth. All results demonstrated that the combination of the components used here as a dermal device, chosen according to previous miscellany studies of low/mid-cost biomaterials, can promote skin protection avoiding infections and dehydration, minimize the typical wound inflammatory responses, and favor the cellular healing responses, features that give rise to further clinical trials of the device here developed.

Thermal degradation of calcium and sodium alginate: A greener synthesis towards calcium oxide micro/nanoparticles.

Processes for nanoparticle synthesis often use toxic solvents under aggressive conditions. A greener alternative is the burning of self-organized alginate systems. We followed the influence of the CaCl2 concentrations during gelation of sodium alginate and the heating rate on the synthesis of nanoparticles by the combustion method using TGA as a reactor vessel. Samples were collected after each main process of mass loss and characterized using the Scanning Electron Microscopy, Infrared Spectroscopy, and X-ray Diffraction. Samples treated at 50 °C·min-1 presented porous structures at temperatures more than 500 °C lower than the treatments at 10 °C·min-1. All calcium alginate samples presented changing from a predominantly amorphous to crystalline structures such as Ca(OH)2, CaCO3 in the calcite phase and CaO as a function of the temperature, while sodium alginate produced mainly Na2CO3, NaOH and NaO. We observed two main correlations: 1) between the porosity and the heating rate, and 2) between the formation of crystalline structure in intermediate temperatures and the CaCl2 concentration in the gelation step.

Cross-reactivity between myelin oligodendrocyte glycoprotein and human endogenous retrovirus W protein: nanotechnological evidence for the potential trigger of multiple sclerosis.

Multiple sclerosis (MS) is an autoimmune and inflammatory demyelinating disease of the central nervous system. Experimental evidence supports the reactivity of autoantibodies against components of myelin sheath including the myelin oligodendrocyte glycoprotein (MOG). The MS etiology is still unknown, but some risk factors associated with immune dysregulation, genetic susceptibility, and environmental factors are under investigation. The last consider the hypothesis of molecular mimicry mechanism, which is potentially triggered by viral antigen inducing MS autoimmunity. The Human Endogenous Retroviruses W family (HERV-W) is the subject of studies within this field, based on the detection of HERV-W envelope gene proteins in MS patients' samples. In the biomedical field of diagnosis and therapeutics, nanotechnology is of great use for the detailed study of molecular mechanisms involving specific interactions between biomolecules providing high specificity and sensitivity of response. In view of the significance of etiological aspects for the comprehension of MS mechanisms of action, we applied a nanotechnological approach designed for antibody detection. For this, we analyzed MOG peptide sequences similar to the HERV-W protein. These sequences were subjected to interaction with anti-HERV-W antibodies using atomic force spectroscopy (AFS) and silver nanoparticles (AgNPs) methods to survey the potential occurrence of molecular mimicry. Our results revealed the molecular recognition between the anti-HERV-W antibody and the HERV-W and MOG epitopes by AFS and AgNPs approaches. Specific non-linear shape of force curves and median adhesion force values within the expected range for an antigen-antibody interaction were obtained for HERV-W and MOG peptides, 163 pN and 178 pN, respectively, suggesting the occurrence of cross-reactivity in these systems.

Peptide-Conjugated Silver Nanoparticle for Autoantibody Recognition.

In this work, we considered the autoantibodies proposed as putative biomarkers of demyelination taking into account their reactivity towards myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP). These myelin proteins are among the most commonly researched targets in the immunopathology of demyelinating diseases. In this context, the development of assays for autoantibody detection can contribute as a predictive value for the early diagnosis of demyelinating diseases. Hence, we aimed to address the application of silver nanoparticles (AgNPs) as a sensing device of autoantibodies. AgNPs were synthesized via a chemical reduction method and characterized using atomic force microscopy (AFM), X-ray diffractometry, dynamic light scattering, and UV-visible spectrophotometry. The process of peptide conjugation on the nanoparticles was also analyzed. The autoantibody recognition by the peptide-conjugated AgNPs was evaluated with UV-visible spectrophotometry, atomic force spectroscopy (AFS), and color changing. AgNPs exhibited spherical morphology, low polydispersity, face-centered cubic crystal structure, and an average size of 29.3±3.0 nm. The hydrodynamic diameter variation and AFM showed that the MBP peptide induced greater agglomeration, compared to MOG peptide. The AFS measurements indicated the efficient binding of peptides to the AgNPs maintaining their activity, revealed by typical adhesion force and shapes of curves. The absorption spectrum features were more affected by the interaction with the specific autoantibodies, which also caused a visible color change in suspension providing a qualitative response. We described a preliminary study of MOG- and MBP-conjugated AgNPs which showed to be applicable in the autoantibody recognition. These have promising implication in the searching for biological markers for diagnostic purposes in the demyelination context, in which the nanoscale sensing exploitation is recent.

A Study of the Pharmacokinetic Properties and the In Vivo Kinetics of Erythrocytes Loaded With Dexamethasone Sodium Phosphate in Healthy Volunteers.

The objectives of this 2-phase study were to elucidate pharmacokinetics (PK), in vivo 24-hour recovery, and red blood cell (RBC) survival properties of RBC-encapsulated dexamethasone sodium phosphate (DSP) prepared using the EryDex System (EDS). The 24-hour RBC recovery and T50 survival phase studied subjects were randomized to receive autologous RBCs loaded with either 15-20 mg DSP (Group 1A) or sham saline (Group 2A). Loaded RBCs were radiolabeled with 51-Cr, and the labeled RBCs were followed over time in vivo. The PK phase evaluated dose levels of 2.5-5 mg (Group 1B) and 15-20 mg (Group 2B) DSP encapsulated in RBCs infused into healthy randomized subjects. The mean ± SD 24-hour RBC recovery was 77.9% ± 3.3% and 72.7% ± 10.5% for Groups 1A and 2A, respectively. The mean ± SD RBC life span was 84.3 ± 8.3 days in Group 1A and 88.9 ± 6.2 days in Group 2A. The PK phase actual DSP loading doses (mean ± SEM) were 4.2 ± 0.27 mg and 16.9 ± 0.90 mg in Groups 1B and 2B, respectively. Release of dexamethasone from RBCs in vivo peaked at 1 hour, and a sustained release of dexamethasone could be detected until 35 days after the single intravenous infusion in Group 2B. The mean RBC in vivo recovery for DSP-loaded processed cells compares similarly to the 24-hour recovery of regulated RBC products intended for transfusion. There was a minimal but acceptable adverse impact on the survival of EDS-processed RBCs. DSP-loaded autologous RBCs, prepared using the EDS, delivered a sustained dose of dexamethasone in vivo.

Ex vivo encapsulation of dexamethasone sodium phosphate into human autologous erythrocytes using fully automated biomedical equipment.

Erythrocyte-based drug delivery systems are emerging as potential new solutions for the release of drugs into the bloodstream. The aim of the present work was to assess the performance of a fully automated process (EDS) for the ex-vivo encapsulation of the pro-drug dexamethasone sodium phosphate (DSP) into autologous erythrocytes in compliance with regulatory requirements. The loading method was based on reversible hypotonic hemolysis, which allows the opening of transient pores in the cell membrane to be crossed by DSP. The efficiency of encapsulation and the biochemical and physiological characteristics of the processed erythrocytes were investigated in blood samples from 34 healthy donors. It was found that the processed erythrocytes maintained their fundamental properties and the encapsulation process was reproducible. The EDS under study showed greater loading efficiency and reduced variability compared to previous EDS versions. Notably, these results were confirmed using blood samples from Ataxia Telangiectasia (AT) patients, 9.33±1.40 and 19.41±2.10mg of DSP (mean±SD, n=134) by using 62.5 and 125mg DSP loading quantities, respectively. These results support the use of the new EDS version 3.2.0 to investigate the effect of erythrocyte-delivered dexamethasone in regulatory trials in patients with AT.