Aceite de cáscara de citrus reticulata con potencial terapéuticos para el tratamiento de la aterosclerosis : estudios bioquímicos, moleculares y bioinformáticos

La aterosclerosis es una enfermedad cardiovascular (ECV) caracterizada por un engrosamiento de las paredes arteriales debido al depósito de lípidos principalmente el colesterol (Col) y a una respuesta inflamatoria crónica promovida por macrófagos y células espumosas. Las ECVs son la principal causa de muerte y discapacidad a nivel mundial siendo la hipercolesterolemia el factor que contribuye al 56% de los casos de cardiopatía coronaria. Los niveles de Col plasmático se regulan por mecanismos como la síntesis de novo del Col o vía del mevalonato (VM) principalmente activa en hepatocitos. En las primeras etapas de la VM (reacciones pre-escualeno) se generan isoprenoides no esteroideos como ubiquinona, dolicol y grupos prenilos.Fil: Castro, M.. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Llanos, M.. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Rodenak-kladniew, B.. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Gavernet, L. Consejo Nacional de Investigaciones Científicas y TécnicasFil: García de Bravo, M. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Crespo, R.. Consejo Nacional de Investigaciones Científicas y Técnica

Design, Characterization And In Vitro Evaluation Of Linalool-loaded Solid Lipid Nanoparticles As Potent Tool In Cancer Therapy

Linalool (LN) is a monoterpene found in essential oils of plants and herbs that produces multiple effects on the mevalonate pathway and interesting antiproliferative activity in cancer cells. However, due to its poor aqueous solubility, an efficient vehicle is needed to improve its administration and bioavailability in physiological media. LN encapsulation in solid lipid nanoparticles (SLN) with different compositions was explored and in vitro tested in two cancer cell lines. SLN of myristyl myristate (MM), cetyl esters (SS) and cetyl palmitate (CP) were prepared by sonication in the presence of Pluronic®F68 as surfactant. Nanoparticle size, morphology and distribution were determined by dynamic light scattering in combination with optical and transmission electron microscopy (TEM). SLN showed spherical shape and mean diameters in the range of 90–130 nm with narrow size dispersion (PDI values lower than 0.2) and Z potentials around −4.0 mV. The encapsulation percentages of LN in SLN were higher than 80% for all tested formulations and exhibited in vitro LN controlled release profiles for at least 72 h. The nanoparticles were physicochemically characterized by FTIR, XRD, DSC and TGA, and the incorporation of LN into SLN was higher than 80% in tested matrices. The developed formulations, and in particular SLN (MM)-LN, showed in vitro antiproliferative effects on hepatocarcinoma (HepG2) and lung adenocarcinoma (A549) cell lines in a dose-dependent response, and higher inhibitory effects were found in comparison with free LN. The cellular uptake of SLN was demonstrated by fluorescence microscopy, enhancing the ability of nanoparticles to intracellularly deliver the cargo molecules. © 2017 Elsevier B.V.15412313211/X701, UNLP, Universidad Nacional de La PlataPRH 5.2, UNLP, Universidad Nacional de La Plat

Effect of geraniol on rat cardiomyocytes and its potential use as a cardioprotective natural compound

Aims Reactive oxygen species (ROS) are generated in the ischaemic myocardium especially during early reperfusion and affect myocardial function and viability. Monoterpenes have been proposed to play beneficial roles in diverse physiological systems; however, the mechanisms of action remain largely unknown. This study aims to assess the effect of monoterpene geraniol (GOH) on neonatal rat ventricular cardiomyocytes (NRVCs) subjected to oxidative stress. Main methods We used an in vitro model of hypoxia-reoxygenation. Cardioprotective (AMPK) and cardiotoxic (ERK1/2, ROS) signaling indicators were measured. Assays were performed by fluorogenic probes, MTT assays and Western-blots. Key findings We determined that the addition of GOH (5–200 μM) to cultured normoxic and hypoxic-NRVCs diminished the endogenous production of ROS in stressed cardiomyocytes. We observed that GOH treatment increased pAMPK levels and decreased pERK1/2 levels in cultured NRVCs. Significance This report suggests that GOH is a candidate cardioprotective natural compound that operates by blunting the oxidative stress signaling that is normally induced by hypoxia-reoxygenation

Origen y distribución espacial de linajes maternos nativos en el noroeste y centro oeste argentinos/ Origin and spatial distribution of native maternal lineages in Northwest and Center West of Argentina

RESUMEN El empleo de los polimorfismos del ADN mitocondrial para caracterizar a las poblaciones humanas ha permitido distinguir patrones geográficos de distribución de linajes, detectando posibles rutas de poblamiento. En Argentina existen regiones clásicamente identificadas como libres de pueblos originarios, quedando espacios vacíos de información en cuanto a la distribución de linajes nativos. Sin embargo, los pueblos de las regiones del centro-oeste y noroeste no fueron exterminados sino que fueron asimilados biológicamente primero, a la sociedad colonial y luego, a la sociedad estatal. Los procesos de mestizaje tuvieron una tendencia sexo asimétrica permitiendo la conservación de los linajes maternos nativos en elevadas proporciones. En este trabajo se analizan 1951 muestras obtenidas en centros de salud de catorce localidades del centro-oeste y noroeste de Argentina y se comprueba que el 90% de los haplogrupos mitocondriales son propios de América. La agrupación de localidades en base a la distribución de frecuencias de haplogrupos nativos mediante AMOVA indica que las localidades de Maimará y La Quiaca constituyen una entidad claramente diferenciada, en coincidencia con los datos arqueológicos y lingüísticos. Las elevadas frecuencias de haplogrupo D en La Rioja y de haplogrupo A en Villa Tulumaya pueden representar fenómenos poblacionales prehispánicos. Se postula entonces la utilidad de abordar a la población actual como vía de acceso al conocimiento del origen de los linajes mitocondriales nativos y su distribución espacial.  PALABRAS CLAVE ADN mitocondrial; nativos americanos; mestizaje ABSTRACT The use of mitochondrial DNA polymorphisms for identifying human populations has allowed the distinction of geographical patterns of lineage distribution, detecting possible peopling routes. In Argentina there are regions that were classically defined as empty of Native American people, leaving areas without information regarding the Native lineage distribution. However, the peoples of the Center-West and North-West were not exterminated but biologically assimilated first to the colonial society and then to the state society. The admixture processes had a sex-asymmetric tendency, allowing the conservation of maternal lineages in high proportions. In this paper we analyze 1951 samples from healthcare centers in 14 locations of the Center-West and North-West of Argentina and we found that 90% of the mitochondrial haplogroups are American. This means that by studying the current population we can access the Native American mitochondrial lineages. In this context we tested grouping locations by their haplogroup frequency distribution with an AMOVA. We found that Maimará and La Quiaca are a clearly differentiated entity, in coincidence with archaeological and linguistic data. It should be pointed out that the high frequencies of haplogroup D in La Rioja and haplogroup A in Villa Tulumaya may represent prehispanic population phenomena. We hypothesize then the usefulness of studying the current population as a way to know the origin of native mitochondrial lineages and their spatial distribution.  KEY WORDS mitochondrial DNA; Native Americans; miscegenatio

Origen y distribución espacial de linajes maternos nativos en el noroeste y centro oeste argentinos/ Origin and spatial distribution of native maternal lineages in Northwest and Center West of Argentina

RESUMEN El empleo de los polimorfismos del ADN mitocondrial para caracterizar a las poblaciones humanas ha permitido distinguir patrones geográficos de distribución de linajes, detectando posibles rutas de poblamiento. En Argentina existen regiones clásicamente identificadas como libres de pueblos originarios, quedando espacios vacíos de información en cuanto a la distribución de linajes nativos. Sin embargo, los pueblos de las regiones del centro-oeste y noroeste no fueron exterminados sino que fueron asimilados biológicamente primero, a la sociedad colonial y luego, a la sociedad estatal. Los procesos de mestizaje tuvieron una tendencia sexo asimétrica permitiendo la conservación de los linajes maternos nativos en elevadas proporciones. En este trabajo se analizan 1951 muestras obtenidas en centros de salud de catorce localidades del centro-oeste y noroeste de Argentina y se comprueba que el 90% de los haplogrupos mitocondriales son propios de América. La agrupación de localidades en base a la distribución de frecuencias de haplogrupos nativos mediante AMOVA indica que las localidades de Maimará y La Quiaca constituyen una entidad claramente diferenciada, en coincidencia con los datos arqueológicos y lingüísticos. Las elevadas frecuencias de haplogrupo D en La Rioja y de haplogrupo A en Villa Tulumaya pueden representar fenómenos poblacionales prehispánicos. Se postula entonces la utilidad de abordar a la población actual como vía de acceso al conocimiento del origen de los linajes mitocondriales nativos y su distribución espacial.  PALABRAS CLAVE ADN mitocondrial; nativos americanos; mestizaje ABSTRACT The use of mitochondrial DNA polymorphisms for identifying human populations has allowed the distinction of geographical patterns of lineage distribution, detecting possible peopling routes. In Argentina there are regions that were classically defined as empty of Native American people, leaving areas without information regarding the Native lineage distribution. However, the peoples of the Center-West and North-West were not exterminated but biologically assimilated first to the colonial society and then to the state society. The admixture processes had a sex-asymmetric tendency, allowing the conservation of maternal lineages in high proportions. In this paper we analyze 1951 samples from healthcare centers in 14 locations of the Center-West and North-West of Argentina and we found that 90% of the mitochondrial haplogroups are American. This means that by studying the current population we can access the Native American mitochondrial lineages. In this context we tested grouping locations by their haplogroup frequency distribution with an AMOVA. We found that Maimará and La Quiaca are a clearly differentiated entity, in coincidence with archaeological and linguistic data. It should be pointed out that the high frequencies of haplogroup D in La Rioja and haplogroup A in Villa Tulumaya may represent prehispanic population phenomena. We hypothesize then the usefulness of studying the current population as a way to know the origin of native mitochondrial lineages and their spatial distribution.  KEY WORDS mitochondrial DNA; Native Americans; miscegenatio

Origen y distribución espacial de linajes maternos nativos en el noroeste y centro oeste argentinos/ Origin and spatial distribution of native maternal lineages in Northwest and Center West of Argentina

RESUMEN El empleo de los polimorfismos del ADN mitocondrial para caracterizar a las poblaciones humanas ha permitido distinguir patrones geográficos de distribución de linajes, detectando posibles rutas de poblamiento. En Argentina existen regiones clásicamente identificadas como libres de pueblos originarios, quedando espacios vacíos de información en cuanto a la distribución de linajes nativos. Sin embargo, los pueblos de las regiones del centro-oeste y noroeste no fueron exterminados sino que fueron asimilados biológicamente primero, a la sociedad colonial y luego, a la sociedad estatal. Los procesos de mestizaje tuvieron una tendencia sexo asimétrica permitiendo la conservación de los linajes maternos nativos en elevadas proporciones. En este trabajo se analizan 1951 muestras obtenidas en centros de salud de catorce localidades del centro-oeste y noroeste de Argentina y se comprueba que el 90% de los haplogrupos mitocondriales son propios de América. La agrupación de localidades en base a la distribución de frecuencias de haplogrupos nativos mediante AMOVA indica que las localidades de Maimará y La Quiaca constituyen una entidad claramente diferenciada, en coincidencia con los datos arqueológicos y lingüísticos. Las elevadas frecuencias de haplogrupo D en La Rioja y de haplogrupo A en Villa Tulumaya pueden representar fenómenos poblacionales prehispánicos. Se postula entonces la utilidad de abordar a la población actual como vía de acceso al conocimiento del origen de los linajes mitocondriales nativos y su distribución espacial.  PALABRAS CLAVE ADN mitocondrial; nativos americanos; mestizaje ABSTRACT The use of mitochondrial DNA polymorphisms for identifying human populations has allowed the distinction of geographical patterns of lineage distribution, detecting possible peopling routes. In Argentina there are regions that were classically defined as empty of Native American people, leaving areas without information regarding the Native lineage distribution. However, the peoples of the Center-West and North-West were not exterminated but biologically assimilated first to the colonial society and then to the state society. The admixture processes had a sex-asymmetric tendency, allowing the conservation of maternal lineages in high proportions. In this paper we analyze 1951 samples from healthcare centers in 14 locations of the Center-West and North-West of Argentina and we found that 90% of the mitochondrial haplogroups are American. This means that by studying the current population we can access the Native American mitochondrial lineages. In this context we tested grouping locations by their haplogroup frequency distribution with an AMOVA. We found that Maimará and La Quiaca are a clearly differentiated entity, in coincidence with archaeological and linguistic data. It should be pointed out that the high frequencies of haplogroup D in La Rioja and haplogroup A in Villa Tulumaya may represent prehispanic population phenomena. We hypothesize then the usefulness of studying the current population as a way to know the origin of native mitochondrial lineages and their spatial distribution.  KEY WORDS mitochondrial DNA; Native Americans; miscegenatio

Hybrid ofloxacin/eugenol co-loaded solid lipid nanoparticles with enhanced and targetable antimicrobial properties

In the global context of an imminent emergence of multidrug-resistant microorganisms, the present work combined the use of nanotechnology and the therapeutic benefits of natural compounds as a strategy to potentiate antimicrobial action of the wide-spectrum antibiotic Ofloxacin (Ofx). Hybrid solid lipid nanoparticles (SLN) were synthesized by incorporation of chitosan (Chi, a cationic biopolymer with antimicrobial activity) and eugenol (Eu, a phenolic compound that interferes with bacterial quorum sensing) into a lipid matrix by hot homogenization/ultrasonication method. The developed SLN/Chi/Eu sustainably released the encapsulated Ofx for 24 h. Characterization by DLS, TEM, DSC, TGA and XRD revealed the presence of positively charged spherical nanoparticles with diameters around 300 nm and Ofx entrapped in amorphous state. The SLN exhibited an enhanced bactericidal activity against Pseudomonas aeruginosa and Staphylococcus aureus. The minimum inhibitory concentration (MIC) for free and nanoencapsulated Ofx formulations was below 1.0 mu g/ml. The MIC values decreased by 6.1- to 16.1-fold when Ofx was encapsulated in SLN/Chi/Eu. Fluorescent-labeled nanoparticles had the ability to interact with the bacterial cell membrane. Selective toxicity of SLN/Chi/Eu-Ofx was tested in the range of 0.3-30.0 mu g/ml and showed no toxicity up to 3.0 mu g/ml Ofx in human cell models (A549 and Wi-38) at 24 h and 48 h exposure. It was proved that the administration of hybrid SLN to mice by dry powder inhalation reached therapeutic Ofx levels in lungs569The present work was supported by Argentine grants from CONICET (National Council for Science and Technology, PIP 0498), The National Agency of Scientific and Technological Promotion (ANPCyT, PICT 2016-4597), UNLP (Nat. Univ. of La Plata, 12/X545, 13/X545 and 14/X701) and Fundación Argentina de Nanotecnología (FAN) to GRC. Also, GAI wants to thank the Grant for Jóvenes Investigadores UNLP 2017 and Premio a la Innovación UNLP 2014 for financing some reagents. BRK is grateful to UNLP for financial support through the Subsidio para jóvenes investigadores 2016. The technical collaboration of the students Ms. F. Blandin and Ms. C. Marc (University of Angers, France), as well as Mr. N. Noacco and Ms. C. Toledo (UNLP), is also gratefully acknowledged. We are also indebted to Dr. N. Scaglia (UNLP, Argentina) for the Wi-38 cell line donation. Finally, we want to thank CRODA Argentina for kindly donating the lipids, and Mr. A. Torres Nicolini and Ms. M. Bracone for their collaboration on physicochemical characterization