Expresión, localización y estudio funcional de PGRMC1 en el endometrio humano a lo largo del ciclo menstrual

Las acciones de la progesterona (P4), en el sistema reproductivo femenino, están principalmente mediadas por los receptores clásicos de la progesterona (PR), que actúan como factores de transcripción. Sin embargo, no todas las acciones fisiológicas de la progesterona pueden explicarse por la activación de dichos receptores. De hecho, se conoce que la progesterona actúa también a través de vías de señalización no clásica más rápidas, mediadas por la activación de receptores de la membrana celular. Entre los receptores no clásicos de la progesterona encontramos PGRMC1 (Progesterone receptor membrane component 1), cuyo mecanismo de acción queda por ser definido. La interacción P4-PGRMC1 media las acciones antimitóticas y antiapoptóticas en las células de la granulosa ováricas, regulando así el desarrollo folicular. Sin embargo, se sabe poco sobre la implicación de PGRMC1 en el endometrio humano. Esta tesis doctoral se ha centrado en el estudio del receptor no clásico de la P4 (PGRMC1) en el endometrio humano a lo largo del ciclo menstrual, enfocándose especialmente en su función en el proceso de decidualización. Hemos visto que PGRMC1 presenta una expresión variable en el endometrio humano durante el ciclo menstrual. Entre los diferentes compartimentos endometriales, PGRMC1 se expresa mayormente en el compartimento estromal, donde se ha visto que cambia su localización durante el proceso de decidualización, mostrando principalmente una localización perinuclear y nuclear. Gracias a estudios in vitro se ha visto que la sobrexpresión de PGRMC1 inhibe el proceso de decidualización en cultivos primarios de células endometriales estromales humanas (ESC). Por otro lado, en ausencia de la entrada de P4 en las ESC, la activación de los receptores no clásicos de membrana de la P4 y el utilizo de un antagonista de PGRMC1 fueron capaces de inducir los cambios celulares típicos de decidualización. Esto indica que PGRMC1 estaría directamente implicada en el proceso de decidualización. La presencia del antagonista de PGRMC1 durante los estímulos de decidualización generó, abundantes cambios transcriptómicos, con un aumento significativo de todos los procesos asociados a actividad mitocondrial y disminución de actividad de transporte intracelular. Para poder entender mejor a cerca de la función de PGRMC1 durante la decidualización, se ha procedido en la búsqueda de interacciones proteicas en ESC no decidualizadas y decidualizadas. Se han identificado diferentes proteínas especificas que interaccionan con PGRMC1 antes y despúes de la decidualización, localizadas principalmente en lisosomas y mitocondrias y relacionadas con actividad de transporte celular y actividad catalitica. Todo eso indica que PGRMC1 podría desempeñar, en las ESC, un papel de transporte y obtención de energía, procesos biológicos importantes durante la profunda remodelación que sufren las ESC durante la decidualización

Regional diastolic function by tissue Doppler echocardiography in systemic sclerosis: correlation with clinical variables

The incidence of left ventricular (LV) diastolic dysfunction is increased in systemic sclerosis (SSc), while systolic dysfunction is present in a small percentage of patients. The aim of this study was to asses the LV "regional" diastolic abnormalities in SSc patients by the mean of Doppler tissue imaging (DTI). Echocardiographic echo-Doppler (DE) and DTI parameters were analyzed for 67 SSc patients: abnormal E/A ratio at DE was detected in 24, while abnormal e/a at DTI was observed in 41. A significant prevalence of DTI diastolic abnormalities in the segments reflecting longitudinal versus those reflecting radial LV motion was found. The segments of the basal regions of LV myocardium were significantly more involved than those of the middle portion. Linear correlation was observed between the extent of the diastolic abnormalities and the duration of disease. Longitudinal myocardial systolic velocities were significantly reduced in patients with abnormal e/a DTI

Normal and pathogenic variation of RFC1 repeat expansions: implications for clinical diagnosis

Cerebellar Ataxia, Neuropathy and Vestibular Areflexia Syndrome (CANVAS) is an autosomal recessive neurodegenerative disease, usually caused by biallelic AAGGG repeat expansions in RFC1. In this study, we leveraged whole genome sequencing (WGS) data from nearly 10,000 individuals recruited within the Genomics England sequencing project to investigate the normal and pathogenic variation of the RFC1 repeat. We identified three novel repeat motifs, AGGGC (n=6 from 5 families), AAGGC (n=2 from 1 family), AGAGG (n=1), associated with CANVAS in the homozygous or compound heterozygous state with the common pathogenic AAGGG expansion. While AAAAG, AAAGGG and AAGAG expansions appear to be benign, here we show a pathogenic role for large AAAGG repeat configuration expansions (n=5). Long read sequencing was used to fully characterise the entire repeat sequence and revealed a pure AGGGC expansion in six patients, whereas the other patients presented complex motifs with AAGGG or AAAGG interruptions. All pathogenic motifs seem to have arisen from a common haplotype and are predicted to form highly stable G quadruplexes, which have been previously demonstrated to affect gene transcription in other conditions. The assessment of these novel configurations is warranted in CANVAS patients with negative or inconclusive genetic testing. Particular attention should be paid to carriers of compound AAGGG/AAAGG expansions, since the AAAGG motif when very large (>500 repeats) or in the presence of AAGGG interruptions. Accurate sizing and full sequencing of the satellite repeat with long read is recommended in clinically selected cases, in order to achieve an accurate molecular diagnosis and counsel patients and their families

Optimal superfluid management of 5G networks

We consider the problem of evaluating the performance of a 5G network based on reusable components, called Reusable Functional Blocks (RFBs), proposed by the Horizon 2020 SUPERFLUIDITY project. RFBs allow a high level of flexibility, agility, portability and high performance. After formally modelling the RFB entities and the network physical nodes, we optimally formulate the problem of maximizing different Key Performance Indicators (KPIs) on an RFB-based network architecture, in which the RFBs are shared among the nodes, and deployed only where and when they are really needed. Our results, obtained by solving the proposed optimization problem over a simple yet representative scenario, show that the network can be managed in a very efficient way. More in depth, the RFBs are placed into the nodes in accordance with the amount of requested traffic from users and the specific pursued KPI, e.g., maximization of user throughput or minimization of the number of used nodes. Moreover, we evaluate the relationship between the capacity of each node and the number of RFBs deployed on it

iTRAQ comparison of proteomic profiles of endometrial receptivity

Endometrial receptivity is a limiting step in human reproduction. A disruption in the development of endometrial receptivity is responsible for recurrent implantation failures (RIF) of endometrial origin. To understand the molecular mechanisms behind the endometrial receptivity process, we used the isobaric tag for relative and absolute quantitation (iTRAQ) method to compare three different endometrial statuses: fertile women, intrauterine device (IUD) carriers, and RIF patients. Overall, iTRAQ allowed identified 1889 non-redundant proteins. Of these, 188 were differentially expressed proteins (DEP) (p-value < .05). Pairwise comparisons revealed 133 significant DEP in fertile vs. IUD carriers and 158 DEP in RIF vs. IUD carriers. However, no DEP were identified between fertile and RIF patients. Western blot validation of three DEP involved in endometrial receptivity (plastin 2, lactotransferrin, and lysozyme) confirmed our iTRAQ results. Moreover, functional KEGG enrichment revealed that complement and coagulation cascades and peroxisome were the two most significant pathways for the RIF vs. IUD comparison and ribosome and spliceosome for the fertile vs. IUD comparison, as possible important pathways involved in the endometrial receptivity acquisition. The lack of DEP between fertile and RIF patient endometria suggest that idiopathic RIF may not have an endometrial origin, with other as-yet-unknown factors involved