Toll like receptors agonists-based nanomedicines as veterinary immunotherapies

Many infections affecting animals enter across mucosa, needing of secretory immunity to reject the disease; protection against some pathogens must be early, fast, and specifically elicited, while from others must be wide enough to fight against variable serotypes. Today, however, most veterinary vac-cines are injectable (not recommended for chicken and small fishes), aimed to control clinical signs instead of eradicating the disease and poor inducers of secretory immunity. These drawbacks are com-pensated by vaccination with live attenuated agents, by using potentially toxic oily adjuvants and with indiscriminate use of antibiotics. In this review, the benefits of commercial and experimental nanomed-icines acting as immunostimulants and vaccine adjuvants made of toll-like receptor (TLR) agonistsloaded in nanoparticles (NP-TLR agonists), arepresented. Np-TLR agonists induce a magnified, site-limited triggering of innate immunity; also allow modifying the administration route from injectable to mucosal or nebulized, provide structural protection to TLR agonists and avoid their diffusion far from the administration site. Future implementation of immunotherapies based on Np-TLR agonists will be discussed as a function of scale production feasibility.Fil: Parra, Federico Leonel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Diseño de Estrategias de Targeting de Drogas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Morilla, María José. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Diseño de Estrategias de Targeting de Drogas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romero, Eder Lilia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Diseño de Estrategias de Targeting de Drogas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Nanomedicines, the essential is invisible… to the light microscope

El poder de resolución del microscopio óptico es de unos 200 nm y si tenemos suerte, los nano-objetos más voluminosos apenas se distinguirán como puntos fluorescentes. ¿Podremos, mediante el control de la arquitectura de esos “puntos brillantes”, dirigir su acceso a ciertos sectores del cuerpo, donde ejecutarían exitosamente funciones terapéuticas? Por ahora estamos más lejos de lo que creemos en materializar ese ejercicio mental. Sin embargo, de todas las aplicaciones de la Nanotecnología al área de la salud, el campo terapéutico es el más importante. En este artículo nos adentraremos en los fenómenos que explican cómo funcionan los nano-objetos que transportan principios activos hasta los sitios blancos. A partir de allí será posible construir expectativas razonables sobre la irrupción de esta nueva tecnología en el corto futuro. Sus potencialidades y limitaciones son dependientes de nuestra capacidad de diseño racional, de desarrollo industrial y conocimiento de los fenómenos de nanotoxicidad, respectivamente.The resolution power of the optical microscope is nearly 200 nm and if we are lucky, the biggest nano-objects are saw as highly fluorescent dots. How the architecture of these shiny dots be controlled, to target them to specific body sites and successfully exert therapeutic actions? For the moment, we have been unable of meeting such issue. Nonetheless, drug delivery is by far the most important field of all the applications of Nanotechnology in health. In this review, we will address the phenomena underlying the action of nano-objects as drug delivery systems. On that knowledge, it will be possible to build reasonable expectancies to the irruption of these new medicines, in the near future. Potentialities and pitfalls of nanomedicines will depend on our skills for rational design, industrial development and insights gained on the new phenomena of nanotoxicity.Fil: Romero, Eder Lilia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Morilla, María José. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Selective cytotoxicity of PAMAM G5 core-PAMAM G2.5 shell tecto-dendrimers on melanoma cells

Background: The controlled introduction of covalent linkages between dendrimer building blocks leads to polymers of higher architectural order known as tecto-dendrimers. Because of the few simple steps involved in their synthesis, tecto-dendrimers could expand the portfolio of structures beyond commercial dendrimers, due to the absence of synthetic drawbacks (large number of reaction steps, excessive monomer loading, and lengthy chromatographic separations) and structural constraints of high-generation dendrimers (reduction of good monodispersity and ideal dendritic construction due to de Gennes dense-packing phenomenon). However, the biomedical uses of tecto-dendrimers remain unexplored. In this work, after synthesizing saturated shell core-shell tecto-dendrimers using amine-terminated polyamidoamine (PAMAM) generation 5 (G5) as core and carboxyl-terminated PAMAM G2.5 as shell (G5G2.5 tecto-dendrimers), we surveyed for the first time the main features of their interaction with epithelial cells. Methods: Structural characterization of G5G2.5 was performed by polyacrylamide gel electrophoresis, matrix-assisted laser desorption time-of-flight mass spectrometry, and microscopic techniques; their hydrodynamic size and Z-potential was also determined. Cellular uptake by human epidermal keratinocytes, colon adenocarcinoma, and epidermal melanoma (SK-Mel-28) cells was determined by flow cytometry. Cytotoxicity was determined by mitochondrial activity, lactate dehydrogenase release, glutathione depletion, and apoptosis/necrosis measurement. Results: The resultant 60%-67% saturated shell, 87,000-dalton G5G2.5 (mean molecular weight) interacted with cells in a significantly different fashion in comparison to their building blocks and to its closest counterpart, PAMAM G6.5. After being actively taken up by epithelial cells, G5G2.5 caused cytotoxicity only on SK-Mel-28 cells, including depletion of intracellular glutathione and fast necrosis that was manifested above 5 μM G5G2.5. It cannot be discounted that traces of LiCl within G5G2.5 were involved in such deleterious effects. Conclusion: These preliminary results suggest that at concentrations that do not damage healthy keratinocytes, G5G2.5 could display antimelanoma activity.Centro de Investigaciones Cardiovasculare

A Poglut1 mutation causes a muscular dystrophy with reduced Notch signaling and satellite cell loss

Skeletal muscle regeneration by muscle satellite cells is a physiological mechanism activated upon muscle damage and regulated by Notch signaling. In a family with autosomal recessive limbgirdle muscular dystrophy, we identified a missense mutation in POGLUT1 (protein O-glucosyltransferase 1), an enzyme involved in Notch posttranslational modification and function. In vitro and in vivo experiments demonstrated that the mutation reduces Oglucosyltransferase activity on Notch and impairs muscle development. Muscles from patients revealed decreased Notch signaling, dramatic reduction in satellite cell pool and a muscle-specific adystroglycan hypoglycosylation not present in patients’ fibroblasts. Primary myoblasts from patients showed slow proliferation, facilitated differentiation, and a decreased pool of quiescent PAX7+ cells. A robust rescue of the myogenesis was demonstrated by increasing Notch signaling. None of these alterations were found in muscles from secondary dystroglycanopathy patients. These data suggest that a key pathomechanism for this novel form of muscular dystrophy is Notch-dependent loss of satellite cells.Junta de Andalucía PI-0017-201

Increased brain radioactivity by intranasal 32P-labeled siRNA dendriplexes within in situ-forming mucoadhesive gels

Background: Molecules taken up by olfactory and trigeminal nerve neurons directly access the brain by the nose-to-brain pathway. In situ-forming mucoadhesive gels would increase the residence time of intranasal material, favoring the nose-to-brain delivery. In this first approach, brain radioactivity after intranasal administration of 32P-small interference RNA (siRNA) complexed with poly(amidoamine) G7 dendrimers (siRNA dendriplexes) within in situ-forming mucoadhesive gels, was determined. Materials: 32P-siRNA dendriplexes were incorporated into in situ-forming mucoadhesive gels prepared by blending thermosensitive poloxamer (23% w/w) with mucoadhesive chitosan (1% w/w, PxChi) or carbopol (0.25% w/w, PxBCP). Rheological properties, radiolabel release profile, and local toxicity in rat nasal mucosa were determined. The best-suited formulation was intranasally administered to rats, and blood absorption and brain distribution of radioactivity were measured. Results: The gelation temperature of both formulations was 23°C. The PxChi liquid showed non-Newtonian pseudoplastic behavior of high consistency and difficult manipulation, and the gel retained 100% of radiolabel after 150 minutes. The PxCBP liquid showed a Newtonian behavior of low viscosity and easy manipulation, while in the gel phase showed apparent viscosity similar to that of the mucus but higher than that of aqueous solution. The gel released 35% of radiolabel and the released material showed silencing activity in vitro. Three intranasal doses of dendriplexes in PxCBP gel did not damage the rat nasal mucosa. A combination of 32P-siRNA complexation with dendrimers, incorporation of the dendriplexes into PxCBP gel, and administration of two intranasal doses was necessary to achieve higher brain radioactivity than that achieved by intravenous dendriplexes or intranasal naked siRNA. Conclusion: The increased radioactivity within the olfactory bulb suggested that the combination above mentioned favored the mediation of a direct brain delivery.Facultad de Ciencias MédicasCentro de Investigaciones Cardiovasculare

Reparation of an inflamed air-liquid interface cultured a549 cells with nebulized nanocurcumin

The anti-inflammatory, antifibrotic and antimicrobial activities of curcumin (CUR) are missed because of its low solubility in aqueous media, low bioavailability, and structural lability upon oral intake. Soft nanoparticles such as nanoliposomes are not efficient as CUR carriers, since crystalline CUR is expelled from them to physiological media. Nanostructures to efficiently trap and increase the aqueous solubility of CUR are needed to improve both oral or nebulized delivery of CUR. Here we showed that SRA1 targeted nanoarchaeosomes (nATC) [1:0.4 w:w:0.04] archaeolipids, tween 80 and CUR, 155 ± 16 nm sized of −20.7 ± 3.3 z potential, retained 0.22 mg CUR ± 0.09 per 12.9 mg lipids ± 4.0 (~600 µM CUR) in front to dilution, storage, and nebulization. Raman and fluorescence spectra and SAXS patterns were compatible with a mixture of enol and keto CUR tautomers trapped within the depths of nATC bilayer. Between 20 and 5 µg CUR/mL, nATC was endocytosed by THP1 and A549 liquid–liquid monolayers without noticeable cytotoxicity. Five micrograms of CUR/mL nATC nebulized on an inflamed air–liquid interface of A549 cells increased TEER, normalized the permeation of LY, and decreased il6, tnfα, and il8 levels. Overall, these results suggest the modified pharmacodynamics of CUR in nATC is useful for epithelia repair upon inflammatory damage, deserving further deeper exploration, particularly related to its targeting ability.Fil: Altube, María Julia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Diseño de Estrategias de Targeting de Drogas; ArgentinaFil: Caimi, Lilen Ivonne. Fundación Instituto Leloir; ArgentinaFil: Huck Iriart, Cristián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martín. Escuela de Ciencia y Tecnología; ArgentinaFil: Morilla, María José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Diseño de Estrategias de Targeting de Drogas; ArgentinaFil: Romero, Eder Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Diseño de Estrategias de Targeting de Drogas; Argentin

The Values of Olympism in “The Descent of the Sella”: The relationship of the Creator Dionisio de la Huerta and the President of the I.O.C Juan Antonio Samaranch

El objetivo de este trabajo es hacer un recorrido por los aspectos más importantes de la vida de Dionisio de la Huerta, precursor del tenis en España y sobre todo del piragüismo a través del Descenso Internacional del Sella. Este evento deportivo ha logrado llegar a evolucionar desde un origen de competición de carácter regional, para convertirse en internacional, con la participación en varias ocasiones de más de veinte países. Estas cifras de participación solo se logran en campeonatos Continentales, Mundiales o Juegos Olímpicos. El Descenso Internacional del Sella, durante muchos años fue el acontecimiento deportivo español más importante. Repasaremos las características que lo hicieron posible a través de su creador, condecorado con la medalla de los Forjadores del Deporte otorgada por la Generalitat de Cataluña y tres veces con la Medalla al Mérito Deportivo. También analizaremos su relación con Juan Antonio Samaranch, Delegado Nacional de Deportes presidente del Comité Olímpico Internacional, así como los elementos que caracterizan el Descenso Internacional del Sella desde la perspectiva de los festivales olímpicos griegosThe objective of this paper is to make a tour of the most important aspects of the life of Dionisio de la Huerta, precursor of tennis in Spain and especially canoeing through The Descent of the River Sella, which has made it to evolve from a source regional competition, to become international, involving several times over 20 countries, a figure that is only achieved in continental, World championships or Olympic Games, since for many years was the most important Spanish sporting event. We will review the characteristics that made it possible through its creator, decorated with the Medal of the Sports Forgers granted by the Generalitat of Catalonia and 3 times with the Medal of Sports Merit, his relationship with Juan Antonio Samaranch, National Delegate of the Movement and President of the International Olympic Committee and we will analyze the elements of the International Descent of the Sella from the perspective of the Greek Olympic Game

Repair of the Church of La Purisima Concepcion of Huelva (Spain) with Jet-Grouting, for Damages Caused by the Construction of Diaphragm Walls in its Proximity

The Church of La Purísima Concepcion, raised in 1515 as the second parish of Huelva (Spain) has been affected by diverse natural disasters along its history. The soil of Huelva is mainly mud, very soft and highly compressible. In 1999 the works to build a residential building with four plants below the ground level affected the church and several buildings. Horizontal displacements of the sheet pile wall in top, overcame 12 cm. The settlements were higher to 5 cm. The system of struts, with lengths over to 30 meters used to built the pile wall, was not the best one. Also, the diaphragm walls were designed short, and the dimensions of the element diaphragm wall were incorrect (wide 5 meters). The foundation of the church has been repaired using the technology of jet-grouting, with columns of 15 and of 30 meters of depth and 0.75-0.80 m of diameter. Later, it was realized concrete slab to join all the heads of the columns of jet-grouting. Measurements of levelling have been realized to observe the movements of the church structure during the accomplishment of the jet-grouting

Repair of the Church of La Purísima Concepción of Huelva (Spain) with Jet Grouting, for damages caused by the construction of diaphragm walls in its proximity

The Church of La Purísima Concepcion, raised in 1515 as the second parish of Huelva (Spain) has been affected by diverse natural disasters along its history. The soil of Huelva is mainly mud, very soft and highly compressible. In 1999 the works to build a residential building with four plants below the ground level affected the church and several buildings. Horizontal displacements of the sheet pile wall in top, overcame 12 cm. The settlements were higher to 5 cm. The system of struts, with lengths over to 30 meters used to built the pile wall, was not the best one. Also, the diaphragm walls were designed short, and the dimensions of the element diaphragm wall were incorrect (wide 5 meters). The foundation of the church has been repaired using the technology of jet-grouting, with columns of 15 and of 30 meters of depth and 0.75-0.80 m of diameter. Later, it was realized concrete slab to join all the heads of the columns of jet-grouting. Measurements of levelling have been realized to observe the movements of the church structure during the accomplishment of the jet-grouting

Fast Biofilm Penetration and Anti-PAO1 Activity of Nebulized Azithromycin in Nanoarchaeosomes

Azithromycin (AZ) is a broad-spectrum antibiotic with anti-inflammatory and antiquorum sensing activity against biofilm forming bacteria such as Pseudomonas aeruginosa. AZ administered by oral or parenteral routes, however, neither efficiently accesses nor remains in therapeutic doses inside pulmonary biofilm depths. Instead, inhaled nanocarriers loaded with AZ may revert the problem of low accessibility and permanence of AZ into biofilms, enhancing its antimicrobial activity. The first inhalable nanovesicle formulation of AZ, nanoarchaeosome-AZ (nanoARC-AZ), is here presented. NanoARC prepared with total polar archaeolipids (TPAs), rich in 2,3-di-O-phytanyl-sn-glycero-1-phospho-(3′-sn-glycerol-1′-methylphosphate) (PGP-Me) from Halorubrum tebenquichense archaebacteria, consisted of ∼180 nm-diameter nanovesicles, loaded with 0.28 w/w AZ/TPA. NanoARC-AZ displayed lower minimal inhibitory concentration and minimal bactericidal concentration, higher preformed biofilm disruptive, and anti-PAO1 activity in biofilms than AZ. NanoARC penetrated and disrupted the structure of the PAO1 biofilm within only 1 h. Two milliliters of 15 μg/mL AZ nanoARC-AZ nebulized for 5 min rendered AZ doses compatible with in vitro antibacterial activity. The strong association between AZ and the nanoARC bilayer, combined with electrostatic attraction and trapping into perpendicular methyl groups of archaeolipids, as determined by Laurdan fluorescence anisotropy, generalized polarization, and small-angle X-ray scattering, was critical to stabilize during storage and endure shear forces of nebulization. NanoARC-AZ was noncytotoxic on A549 cells and human THP-1-derived macrophages, deserving further preclinical exploration as enhancers of AZ anti-PAO1 activity.Fil: Altube, María Julia. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Martínez, Melina María Belén. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Malheiros, Barbara. Universidade de Sao Paulo; BrasilFil: Maffía, Paulo C.. Universidad Nacional de Quilmes; ArgentinaFil: Barbosa, Leandro R. S.. Universidade de Sao Paulo; BrasilFil: Morilla, María José. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romero, Eder Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentin