\u3cem\u3eIn Vitro\u3c/em\u3e Determination of Potency of Small Molecule Inhibitors of Arp2/3 Complex

Actin is a key protein building block of actin microfilaments, which are constructed and deconstructed in response to cellular signaling pathways to regulate cellular processes such as motility, division, and endocytosis. Arp2/3 Complex is a 7-subunit protein complex that is in involved in cellular construction of branched actin networks, functioning by attaching to the side of a pre-existing actin filament and nucleating a daughter branch. Overexpression of Arp2/3 complex has been linked to the ability of certain metastatic cancers to proliferate. This work describes the synthesis and in vitro biochemical testing of several molecules predicted by computational docking to be inhibitors of Arp2/3 Complex, and therefore of potential interest in clinical applications. A bulk actin polymerization assay is used as the key method to determine the potency of inhibitor candidates. Structure-activity relationships derived from these results are also discussed

Estudio comparativo entre los modelos geométricos de córneas 3D paciente-específico generados a partir de los topógrafos Sirius y MS-39

La córnea es una estructura transparente, avascular y de geometría semiesférica que se encuentra en la parte frontal del ojo. Esta estructura juega un papel crucial en la refracción y transmisión de luz hacia la retina. Diversas afecciones pueden llegar a alterar su morfología lo que conlleva una disminución de la calidad visual de los pacientes. Los topógrafos corneales son unas tecnologías basadas en técnicas de imagen que permiten evaluar la arquitectura corneal. En función de la técnica de imagen en la que se fundamentan, los topógrafos operan con unos algoritmos internos cuyo funcionamiento detallado a menudo es opaco. En el presente estudio, se modela geométricamente la misma córnea sana paciente-específico utilizando los datos (nubes de puntos) generados por los topógrafos Sirius y MS-39. El modelo tridimensional generado está basado en las nubes de puntos de las superficies corneales generadas por los dos topógrafos. El objetivo principal es comparar los dos modelos geométricos de una misma córnea, obtenidos a partir de los datos proporcionados por cada topógrafo para identificar posibles discrepancias en las representaciones geométricas y determinar si dichas variaciones pueden influir en el diagnóstico clínico de enfermedades corneales.Esta publicación se realizó en el marco del proyecto “Desarrollo y validación de un nuevo concepto de caracterización biomecánica-morfofuncional de la córnea” número de referencia DTS21/00103. Este proyecto ha contado con el apoyo del Instituto de Salud Carlos III y cofinanciado por la Unión Europea

Estudio comparativo entre los modelos geométricos de córneas 3D paciente-específico generados a partir de los topógrafos Sirius y MS-39

La córnea es una estructura transparente, avascular y de geometría semiesférica que se encuentra en la parte frontal del ojo. Esta estructura juega un papel crucial en la refracción y transmisión de luz hacia la retina. Diversas afecciones pueden llegar a alterar su morfología lo que conlleva una disminución de la calidad visual de los pacientes. Los topógrafos corneales son unas tecnologías basadas en técnicas de imagen que permiten evaluar la arquitectura corneal. En función de la técnica de imagen en la que se fundamentan, los topógrafos operan con unos algoritmos internos cuyo funcionamiento detallado a menudo es opaco. En el presente estudio, se modela geométricamente la misma córnea sana paciente-específico utilizando los datos (nubes de puntos) generados por los topógrafos Sirius y MS-39. El modelo tridimensional generado está basado en las nubes de puntos de las superficies corneales generadas por los dos topógrafos. El objetivo principal es comparar los dos modelos geométricos de una misma córnea, obtenidos a partir de los datos proporcionados por cada topógrafo para identificar posibles discrepancias en las representaciones geométricas y determinar si dichas variaciones pueden influir en el diagnóstico clínico de enfermedades corneales.Esta publicación se realizó en el marco del proyecto “Desarrollo y validación de un nuevo concepto de caracterización biomecánica-morfofuncional de la córnea” número de referencia DTS21/00103. Este proyecto ha contado con el apoyo del Instituto de Salud Carlos III y cofinanciado por la Unión Europea

cGMP-Dependent Protein Kinase Type I Is Implicated in the Regulation of the Timing and Quality of Sleep and Wakefulness

Many effects of nitric oxide (NO) are mediated by the activation of guanylyl cyclases and subsequent production of the second messenger cyclic guanosine-3′,5′-monophosphate (cGMP). cGMP activates cGMP-dependent protein kinases (PRKGs), which can therefore be considered downstream effectors of NO signaling. Since NO is thought to be involved in the regulation of both sleep and circadian rhythms, we analyzed these two processes in mice deficient for cGMP-dependent protein kinase type I (PRKG1) in the brain. Prkg1 mutant mice showed a strikingly altered distribution of sleep and wakefulness over the 24 hours of a day as well as reductions in rapid-eye-movement sleep (REMS) duration and in non-REM sleep (NREMS) consolidation, and their ability to sustain waking episodes was compromised. Furthermore, they displayed a drastic decrease in electroencephalogram (EEG) power in the delta frequency range (1–4 Hz) under baseline conditions, which could be normalized after sleep deprivation. In line with the re-distribution of sleep and wakefulness, the analysis of wheel-running and drinking activity revealed more rest bouts during the activity phase and a higher percentage of daytime activity in mutant animals. No changes were observed in internal period length and phase-shifting properties of the circadian clock while chi-squared periodogram amplitude was significantly reduced, hinting at a less robust oscillator. These results indicate that PRKG1 might be involved in the stabilization and output strength of the circadian oscillator in mice. Moreover, PRKG1 deficiency results in an aberrant pattern, and consequently a reduced quality, of sleep and wakefulness, possibly due to a decreased wake-promoting output of the circadian system impinging upon sleep

Effects of Glyphosate and its Formulation, Roundup, on Reproduction in Zebrafish (Danio rerio)

This is an open access article that is freely available in ORE or from the publisher's web site. Please cite the published version.Copyright © 2014 American Chemical SocietyRoundup and its active ingredient glyphosate are among the most widely used herbicides worldwide and may contaminate surface waters. Research suggests both Roundup and glyphosate induce oxidative stress in fish and may also cause reproductive toxicity in mammalian systems. We aimed to investigate the reproductive effects of Roundup and glyphosate in fish and the potential associated mechanisms of toxicity. To do this, we conducted a 21-day exposure of breeding zebrafish (Danio rerio) to 0.01, 0.5, and 10 mg/L (glyphosate acid equivalent) Roundup and 10 mg/L glyphosate. 10 mg/L glyphosate reduced egg production but not fertilization rate in breeding colonies. Both 10 mg/L Roundup and glyphosate increased early stage embryo mortalities and premature hatching. However, exposure during embryogenesis alone did not increase embryo mortality, suggesting that this effect was caused primarily by exposure during gametogenesis. Transcript profiling of the gonads revealed 10 mg/L Roundup and glyphosate induced changes in the expression of cyp19a1 and esr1 in the ovary and hsd3b2, cat, and sod1 in the testis. Our results demonstrate that these chemicals cause reproductive toxicity in zebrafish, although only at high concentrations unlikely to occur in the environment, and likely mechanisms of toxicity include disruption of the steroidogenic biosynthesis pathway and oxidative stress.Natural Environment Research Counci