Ultrasonication of insulin-loaded microgel particles produced by internal gelation: impact on particle's size and insulin bioactivity

Alginate-dextran sulfate (ADS) microgel has been used to protect insulin from gastrointestinal attack and as a carrier to promote insulin permeation through intestinal epithelium. The throughput of ADS submicron particles generation by emulsification/internal gelation is limited by its wide size distribution. The aim of this work was to study the recovery protocol influence on ADS particles through the determination of its impact on particles’ size distribution and bioactivity. ADS particles showed a wide and multimodal distribution, characterized by a high aggregation phenomenon. In an attempt to reverse particles’ tendency to aggregate and to homogenize particle size ADS populations were submitted to ultrasonication, while particle size distribution, physical and chemical stability, and the bioactivity of entrapped insulin were investigated. After ultrasonication a narrower particle population shifted to the nanoscale, with higher physical stability and significant insulin bioactivity was obtained. Emulsification internal/gelation followed by ultrasonication constituted a valid strategy to obtain ADS particles at the submicron range, with high stability and without significantly compromising insulin bioactivity, so offering promises, under previously well established conditions, to evaluate impact of ADS particle's size on biopharmaceutical and pharmacokinetics phases

CarbonNanotubesConjugatedwithTriazole-BasedTetrathiafulvalene-TypeReceptorsfor C60Recognition

Fullerene receptors prepared by a twofold CuI -catalyzed azide-alkyne cycloaddition (CuAAC) reaction with -extended tetrathiafulvalene (exTTF) have been covalently linked to singlewalled carbon nanotubes (SWCNTs) and multi-walled carbon nanotubes (MWCNTs). The nanoconjugates obtained were characterized by several analytical, spectroscopic and microscopic techniques (TEM, FTIR, Raman, TGA and XPS), and evaluated as C60 receptors by UV-Vis spectroscopy. The complexation between the exTTF-triazole receptor in the free state and C60 was also studied by UV-Vis and 1 H NMR titrations, and compared with analogous triazole-based tweezer-type receptors containing the electron-acceptor 11,11,12,12-tetracyano-9,10-anthraquinodimethane (TCAQ) and benzene rings instead of exTTF motifs, providing in all cases very similar values for the association constant (log Ka 3.0‒3.1). Theoretical density functional theory (DFT) calculations demonstrated that the enhanced interaction between the host and the guest upon increasing the size of the -conjugated arms of the tweezer is compensated by an increase in the energy penalty needed to distort the geometry of the host to wrap C60

Probing effects of organic solvents on paracetamol crystallization using in silico and orthogonal in situ methods

This work entails efforts to understand effects of solvent choice on paracetamol crystallization. Various techniques have been developed and implemented to study aforementioned. A clear-cut, direct evidence of two-step nucleation mechanism is demonstrated using a bench top Raman spectrometer and a novel method named as OSANO.Polymorphismus ist die Eigenschaft vieler anorganischer und insbesondere organischer Moleküle, in mehr als einer Struktur zu kristallisieren. Es ist wichtig, die Faktoren zu verstehen, die den Polymorphismus beeinflussen, da er viele physikochemische Eigenschaften wie Stabilität und Löslichkeit beeinflusst. Nahezu 80 % der vermarkteten Medikamente weisen Polymorphismus auf. In dieser Arbeit wurde der Einfluss der Wahl des organischen Lösungsmittels auf den Polymorphismus von Paracetamol untersucht und verschiedene Methoden entwickelt und angewandt, um den Einfluss genauer zu verstehen. Es wurde festgestellt, dass Ethanol viel stärker auf Paracetamol-Kristallisation als Methanol wirkt. Nichtgleichgewichts-Molekulardynamiksimulationen mit periodischer, simulierter Abkühlung (Simulated Annealing) wurden verwendet, um Vorläufer der metastabilen Zwischenprodukte im Kristallisationsprozess zu untersuchen. Es wurde festgestellt, dass die Strukturen der Bausteine der Paracetamol-Kristalle durch geometrische Wechselwirkungen zwischen Lösungsmittel und Paracetamol bestimmt werden. Die statistisch häufigsten Bausteine in der Selbstassemblierung definieren die finale Kristallstruktur. Ein speziell angefertigter akustischer Levitator hat die Proben zuverlässig gehalten, wodurch die Untersuchung des Einflusses von Lösungsmitteln ermöglicht, heterogene Keimbildung abgeschwächt und andere Umgebungsfaktoren stabilisiert wurden. Die Kristallisation wurde in diesem Aufbau mit zeitaufgelöster In-situ-Raman-Spektroskopie verfolgt und mit einer neuen Zielfunktion basierenden Methode der nichtnegativen Matrixfaktorisierung (NMF) analysiert. Orthogonale Zeitrafferfotografie wurde in Verbindung mit NMF verwendet, um eindeutige und genaue Faktoren zu erhalten, die sich auf die Spektren und Konzentrationen verschiedener Anteile der Paracetamol-Kristallisation beziehen, die als latente Komponenten in den unbehandelten Daten vorhanden sind.Polymorphism is the property exhibited by many inorganic and organic molecules to crystallize in more than one crystal structure. There is a strong need for understanding the influencing factors on polymorphism, as it is responsible for differences in many physicochemical properties such as stability and solubility. Nearly 80 % of marketed drugs exhibit polymorphism. In this work, we took the model system of paracetamol to investigate the influence of solvent choice on its polymorphism. Different methods were developed and employed to understand the influence of small organic solvents on the crystallization of paracetamol. Non-equilibrium molecular dynamics simulations with periodic simulated annealing were used as a tool to probe the nature of precursors of the metastable intermediates occurring in the crystallization process. Using this method, it was found that the structures of the building blocks of crystals of paracetamol is governed by solvent-solute interactions. In situ Raman spectroscopy was used with a custom-made acoustic levitator to follow crystallization. This set-up is a reliable method for investigating solvent influence, attenuating heterogeneous nucleation and stabilizing other environmental factors. It was established that as a solvent, ethanol is much stronger than methanol in its effect of driving paracetamol solutions to their crystal form. The time-resolved Raman spectroscopy crystallization data was processed using a newly developed objective function based non-negative matrix factorization method (NMF). An orthogonal time-lapse photography was used in conjunction with NMF to get unique and accurate factors that pertain to the spectra and concentrations of different moieties of paracetamol crystallization existing as latent components in the untreated data

Physical Mechanisms of DNA repair: A single molecule perspective

Wuite, G.J.L. [Promotor]Peterman, E.J.G. [Promotor

Pacemaking Neurons in the study of Parkinson’s Disease

Parkinson’s disease is the second most common neurodegenerative disorder with a signiVcant social cost. The disease that develops over years results in signiVcant movement related problems for the aUected. The pathogenesis however is partially understood. Computational approaches are signiVcant in the analysis of events that are multi-factorial. Parkinson’s Disease results from a system failure that leads to severe degeneration in the substantia nigra , a locus in the mid-brain. Traditional approaches tend to focus on isolated sub-components of the pathogenic pathways. However, such an approach may be inadequate to describe the pathogenesis. Substantia nigra neurons function on an expensive energy budget, due to a high level of arborisation and pacemaking activity. Spontaneous oscillations of these neurons are an important feature of motor control. Pacemaking involves the L-type calcium channel, and could impose long-term accumulation of calcium within its organelles. Modelling of this activity is an important part of developing an understanding of the pathogenic process. We develop a mathematical paradigm to describe this activity with a single compartment approach. To develop the mathematical framework we initially identify the components that contribute to the process and investigate an appropriate mathematical representation for the respective components. In the next part, we bring together such representation to develop a model that can reproduce measured data. Global optimisation strategies are adopted to tune important parameters. The model explicitly describes the dynamics of the transmembrane potential with changes in the levels of important cations. The model is veriVed for two major observations in literature regarding its response in the presence of channel blockers. The model is analysed for parameter bifurcation and stability of oscillations. Finally a framework is proposed to extend the model to include aspects of calcium homeostasis

The key action of estradiol and progesterone enables GnRH delivery during gestation in the South American plains vizcacha, Lagostomus maximus

The South American plains vizcacha, Lagostomus maximus, is the only mammal described so far that shows expression of estrogen receptors (ERs) and progesterone receptors (PRs) in gonadotropin-releasing hormone (GnRH) neurons. This animal therefore constitutes an exceptional model for the study of the effect of steroid hormones on the modulation of the hypothalamic-pituitary-ovarian (HPO) axis. By using both in vivo and ex vivo approaches, we have found that pharmacological doses of progesterone (P4) and estradiol (E2) produced an inhibition in the expression of hypothalamic GnRH, while physiological doses produced a differential effect on the pulsatile release frequency or genomic expression of GnRH. Our ex vivo experiment indicates that a short-term effect of E2 modulates the frequency of GnRH release pattern that would be associated with membrane ERs. On the other hand, our in vivo approach suggests that a long-term effect of E2, acting through the classical nuclear ERs-PRs pathway, would produce the modification of GnRH mRNA expression during the GnRH pre-ovulatory surge. Particularly, P4 induced a rise in GnRH mRNA expression and protein release with a decrease in its release frequency. These results suggest different levels of action of steroid hormones on GnRH modulation. We conclude that the fine action of E2 and P4 constitute the key factor to enable the hypothalamic activity during the pregnancy of this mammal.Fil: Inserra, Pablo Ignacio Felipe. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Charif, Santiago Elías. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Fidel, Victoria. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Giacchino, Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Schmidt, Alejandro Raúl. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Villarreal, Federico M.. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Proietto, Sofia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Cortasa, Santiago Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Corso, María Clara. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gariboldi, María Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Leopardo, Noelia Paola. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fraunhoffer, Nicolás A.. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Di Giorgio, Noelia Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Lux Lantos, Victoria A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Halperin, Julia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Vitullo, Alfredo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; ArgentinaFil: Dorfman, Verónica Berta. Universidad Maimónides. Área de Investigaciones Biomédicas y Biotecnológicas. Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y de Diagnóstico; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin