Aislamiento y cultivo de células osteoblásticas: Interés para la investigación en cirugía ortopédica y traumatología

Describimos un método de aislamiento y caracterización de células osteoblásticas procedentes de la superficie de trabéculas óseas de explantes obtenidos de metafisis de fémures y tibias de conejo y de cabezas femorales humanas de pacientes sometidos a cirugía protésica de cadera. Después de eliminar las células de la médula ósea de los explantes, estos se despositaron sobre dos tipos de mallas, de 80 y 100 µm., obteniéndose un cultivo en monocarpa que se mostró confluente a partir de los 20 días. La población celular aislada presentó un fenotipo osteoblástico utilizando métodos colorimétricos para determinación de la actividad de fosfatasa alcalina (FA) y morfológicos por medio de la observación con microscopía electrónica de barrido. Las células cultivadas exhibieron una morfología diferente según el grado de confluencia de cultivo, siendo en todo momento positivas las sucesivas determinaciones colorimétricas de FA realizadas a lo largo de la experiencia. Consideramos el método de gran interés en investigación en ortopédica, pues permite el estudio de comportamiento de estas células cuando están sometidas a cualquier tipo de estímulo externo, así como cuando se enfrentan a nuevas generaciones de biomateriales, etc.We describe a method of isolation and characterization of osteoblastic cells taken from the surface of bone trabeculae of explants obtained from femur and tibia metaphysis in rabbits and human femoral head in patients undergoing orthopaedic surgery. Afther removing the cells from the bone marrow of explants, they were placed over two mesh, 80 and 100 µmn, obtaining then a culture in monolayer which showed to be confluent after 20 days. The isolated cell population showed an osteoblastic phenotype using colourmetric methods to determinate the alkaline phosphatase (AF) activity and morphologic methods by means of scanning. The cultivated cell showed a different morphology according to the degree of confluence of the culture, being always positive the sucesive colourmetric determinations of AF along the experiment. We consider the method of a great interest for orthopaedic investigations, since it allows the study of the behaviour of these cells when they are subjet to different types of external stimulus as well as meet they the new generations of biomaterials

Influence of the starting day of luteal phase stimulation on double stimulation cycles

BackgroundDouble ovarian stimulation is one of the most used strategies in poor-prognosis patients. There is a high heterogeneity between the studies regarding the execution of this stimulation protocol. The aim of this study was to investigate whether the day on which luteal phase stimulation begins after the first oocyte retrieval affects ovarian response in DuoStim cycles.MethodsThis observational and retrospective study included 541 DuoStim cycles between January 2018 and December 2021 in a private fertility clinic. Patients were assigned to 4 groups according to the timing of the onset of luteal phase stimulation after oocyte retrieval (0-2nd day, 3rd day, 4th day and 5th-6th day). The primary outcome was the number of oocytes retrieved in the luteal phase in each group.ResultsNo differences were found between groups in the number of oocytes collected (5.12 ± 3.56 vs. 5.39 ± 3.74 vs. 5.61 ± 3.94 vs. 5.89 ± 3.92; p=0,6), MII or number of follicles. An increase in the duration of stimulation was found when stimulation started on the 4th day (10.42 ± 2.31 vs. 10.68 ± 2.37 vs. 11.27 ± 2.40 vs. 10.65 ± 2.37 days, p=0,033). A lower number of fertilized oocytes was observed when stimulation began before the fourth day (3.36 ± 2.80 vs. 3.95 ± 2.53 vs. 4.03 ± 2.73 vs. 4.48 ± 3.11; p=0,036). The number of blastocysts was higher when the stimulation started 5-6 days after retrieval (1.82 ± 1.74 vs. 2.13 ± 1.61 vs. 2.33 ± 2.06 vs. 2.91 ± 2.39; p= 0,030).DiscussionThe number of oocytes retrieved does not differ depending on the day that stimulation begins. However, oocytes competence in terms of fertilized oocytes and blastulation, appears to be lower when the second stimulation starts before the fourth day after oocyte retrieval

Alibava : A portable readout system for silicon microstrip sensors

A portable readout system for silicon microstrip sensors is currently being developed. This system uses a front-end readout chip, which was developed for the LHC experiments. The system will be used to investigate the main properties of this type of sensors and their future applications. The system is divided in two parts: a daughter board and a mother board. The first one is a small board which contains two readout chips and has fan-ins and sensor support to interface the sensors. The last one is intended to process the analogue data that comes from the readout chips and from external trigger signals, to control the whole system and to communicate with a PC via USB. The core of this board is a FPGA that controls the readout chips, a 10 bit ADC, an integrated TDC and an USB controller. This board also contains the analogue electronics to process the data that comes from the readout chips. There is also provision for an external trigger input (e.g. scintillator trigger) and a 'synchronised' trigger output for pulsing an external excitation source (e.g. laser system)

Modelado de Procesos de Neurorrehabilitación

La Neurorrehabilitación es un proceso clínico que se centra en el abordaje de la alteración del sistema nervioso. Existe una enorme variabilidad tanto en la tipología como en el grado de las lesiones neurológicas, lo que la convierte en un proceso extremadamente complejo de analizar y comprender. El presente trabajo se centra en el modelado de las principales actividades que se llevan a cabo en el contexto de la Neurorrehabilitación actual con el objetivo de detectar aquellos puntos en que puedan ser mejoradas, tanto a nivel organizativo como a nivel de ejecución. Por otra parte, se trata de comprenderlas en profundidad para tratar de transformarlas posteriormente en nuevas actividades automatizadas y monitorizadas que se ajusten al nuevo paradigma de rehabilitación ubicua, personalizada y basada en la evidencia

ATLAS silicon module assembly and qualification tests at IFIC Valencia

ATLAS experiment, designed to probe the interactions of particles emerging out of proton proton collisions at energies of up to 14 TeV, will assume operation at the Large Hadron Collider (LHC) at CERN in 2007. This paper discusses the assembly and the quality control tests of forward detector modules for the ATLAS silicon microstrip detector assembled at the Instituto de Fisica Corpuscular (IFIC) in Valencia. The construction and testing procedures are outlined and the laboratory equipment is briefly described. Emphasis is given on the module quality achieved in terms of mechanical and electrical stability.Comment: 23 pages, 38 EPS figures, uses JINST LaTeX clas

La Piedra de Novelda: una roca muy utilizada en el patrimonio arquitectónico

Novelda Stone is a calcareous rock, of the biocalcarenite type. It is extracted from the «Vinalopó Medio» area (Alicante, eastern Spain). Novelda stone is known as construction material from the 13th century. From final of the 19th century until the present time, it has been used in Madrid, Valencia and Alicante. In the past, the main quarries from which this type of rock was extracted were located in the municipal districts of Monóvar (Almorquí Stone), Sax (Portazgo Stone) and Elda (Bateig Stone). Nowadays, the main active extraction site is situated in the Bateig Hill quarries. The rock extracted is known commercially as Bateig Stone. The main varieties of this Bateig Stone are: White, Blue, Layer and Fantasy. Each variety of Novelda stone has different durability rate and different pathology depending on their characteristics, particularly on their hydric behaviour patterns. Layer Bateig and Portazgo stone undergo granular disintegration and alveolization and are less durable than the rest. White and Blue Bateig, as well as Almorquí stone, only develop fissures or scaling under extreme conditions.La Piedra de Novelda es una roca carbonática del tipo biocalcarenita. Se extrae del área de Vinalopó Medio (Alicante, España oriental). La piedra de Novelda es conocida como material de construcción desde el siglo XIII. Desde finales del siglo XIX hasta la actualidad, se ha utilizado en edificios y monumentos de Madrid y Valencia, así como Alicante. En el pasado, las canteras principales donde se extrajo este tipo de piedra se localizaban en los distritos municipales de Monóvar (Piedra Almorquí), Sax (Piedra Portazgo) y El da (Piedra Bateig). Hoy en día, la actividad principal se sitúa en las canteras de Bateig. La piedra extraída es conocida comercialmente como Piedra Bateig. Las variedades principales de esta Piedra Bateig son: Blanco, Azul, Llano y Fantasía. Consecuencia de las diferentes propiedades y en especial de los diferentes comportamientos hídricos, la durabilidad de los distintos tipos de Piedra de Novelda y sus patologías son diferentes. La Bateig LLano y la Piedra de Portazgo sufren desagregación granular y alveolización y su durabilidad es menor que la del resto de las variedades. La Bateig Blanco y Bateig Azul, junto con la Piedra Almorquí, sólo en condiciones muy extremas llegan a desarrollar fisuraciones y/o descamaciones

Late Mesolithic burials at Casa Corona (Villena, Spain): direct radiocarbon and palaeodietary evidence of the last forager populations in Eastern Iberia

Current knowledge about the Mesolithic-Neolithic transition in the Central and Western Mediterranean European regions is deeply limited by the paucity of Late Mesolithic human osteological data and the presence of chronological gaps covering several centuries between the last foragers and the first archaeological evidence of farming peoples. In this work, we present new data to fill these gaps. We provide direct AMS radiocarbon dating and carbon (d13C) and nitrogen (d15N) stable isotope analysis were carried out on bone collagen samples of two single burials from the recently discovered open-air Late Mesolithic site of Casa Corona (Villena, Spain). The results shed new light on the chronology and subsistence patterns of the last Mesolithic communities in the Central Mediterranean region of the Iberian Peninsula. Radiocarbon results date the human remains and funerary activity of the site to 6059e5849 cal BC, statistically different from other Late Mesolithic sites and the earliest Neolithic contexts, and bridging the 500 yrs chronological gap of the Mesolithic-Neolithic transition from the area. Isotopic evidence shows that diet was based on terrestrial resources despite the proximity to the site of lagoon and marine ecosystems. This and previous isotope studies from the region suggest a lower reliance upon marine resources than for Atlantic and Cantabrian sites, although intra-regional patterns of neighbouring Mesolithic populations exhibit both fully terrestrial diets and diets with significant amounts of aquatic resources in them. We hypothesize that in the Central Mediterranean region of Spain the Late Mesolithic dietary adaptations imposed structural limits on demographic growth of the last foragers and favoured rapid assimilation by the earliest Neolithic populations

Chemical factors controlling the steady-state distribution of mixed carbonate-siliciclastic sediments in Bayona Bay (northwest Spain)

The relative space distributions of single mineral constituents in mixed terrigenous-carbonated sediments of the Bayona Bay have been analysed. In order to determine the existence of different mixing mechanisms, a study comparing the general sedimentary trends resulting from the average patterns and the single mineral behaviour, reflected in the single-mineral mapping, was carried out. The relative abundance of all mineral phases in surface sediment samples was first determined by quantitative X-ray powder diffraction data, using the Rietveld method. This procedure enabled us to create specific maps for both the minerals forming the terrigenous fraction and the different calcium carbonate polymorphs involved in the carbonatic component. A cross-linked behaviour between the carbonate facies and the terrigenous minerals involved in the calcium carbonate generation was found, suggesting the existence of a chemical control mechanism which, through dissolution-crystallization processes, constrains the mixed environment's long-term evolution. In the general framework of the region studied, these relations operate for the relative amount of plagioclase to calcium carbonate. The existence of dissolution and growth between both minerals, favoured by the high rate of removal, is proposed, yielding a final crossed pattern, representative of the steady-state. Furthermore, a specific analysis, restricted to the region of low rate of material removal and maximum amount of intermixed sediments, clearly shows an anisotropic distribution for every calcium carbonate polymorph (calcite and aragonite). These correlate with the biotite distribution, and can be connected to local variations in seawater Mg/Ca ratio, induced by biotite weathering.En este trabajo se analiza de forma específica la distribución espacial de cada uno de los minerales involucrados en sedimentos mixtos terrígenos-carbonatados de la bahía de Bayona y se compara con la distribución global de sedimentos, tal como resulta de análisis granulométricos, con el fin de determinar la existencia de diferentes mecanismos de mezcla entre los minerales de ambas fracciones. El contenido porcentual en peso de cada fase mineral, en muestras superficiales de sedimento, se determinó a partir de datos cuantitativos de DRX, empleando el método de Rietveld. Este procedimiento nos ha permitido realizar mapas específicos de distribución, tanto para cada mineral de la fracción terrígena como para cada uno de los diferentes polimorfos de carbonato cálcico en la componente carbonatada. Como resultado de este estudio comparativo, se ha encontrado una correlación, en la distribución espacial y granulométrica, de las facies carbonatadas y los minerales terrígenos implicados en la generación de carbonato cálcico. Un posterior análisis de este resultado sugiere la presencia de mecanismos químicos de control que regulan la interacción entre siliciclastos y carbonatos mediante procesos de disolución-cristalización mediados por el agua del mar. El estudio se ha realizado a dos escalas. En el ámbito general de la bahía de Bayona, las relaciones de distribución observadas entre plagioclasas y carbonato cálcico pueden ser explicadas, dentro del balance global del Ca³⁺ disuelto en el agua del mar, mediante la meteorización química de las plagioclasas y la consiguiente generación de carbonatos. En este caso, los patrones de distribución reflejan la aproximación a un estado estacionario mediante sucesivos eventos de mezcla asociados a la alta removilización material en el medio, tanto para la distribución porcentual en peso como para la distribución granulométrica entre ambos minerales. Finalmente, se realiza un análisis particularizado en una región con baja tasa de removilización y donde la interacción entre sedimentos de ambas fracciones es máxima. El resultado muestra claramente una distribución espacial anisótropa para cada uno de las fases de carbonato cálcico (calcita y aragonito). La abundancia relativa de cada uno de los polimorfos, así como el contenido en Mg³⁺ de la calcita, ha sido correlacionado con la distribución de la biotita y explicado a partir de las diferencias de solubilidad en función de la alta relación Mg³⁺ /Ca³⁺ en el agua de mar que resulta, localmente, de la meteorización química de dicho mineral.Instituto Español de Oceanografí

Unmasking a new prognostic marker and therapeutic target from the GDNF-RET/PIT1/p14ARF/p53 pathway in acromegaly

Background: Acromegaly is produced by excess growth hormone secreted by a pituitary adenoma of somatotroph cells (ACRO). First-line therapy, surgery and adjuvant therapy with somatostatin analogs, fails in 25% of patients. There is no predictive factor of resistance to therapy. New therapies are investigated using few dispersed tumor cells in acute primary cultures in standard conditions where the cells do not grow, or using rat pituitary cell lines that do not maintain the full somatotroph phenotype. The RET/PIT1/p14ARF/p53 pathway regulates apoptosis in normal pituitary somatotrophs whereas the RET/GDNF pathway regulates survival, controlling PIT1 levels and blocking p14ARF (ARF) and p53 expression. Methods: We investigated these two RET pathways in a prospective series of 32 ACRO and 63 non-functioning pituitary adenomas (NFPA), studying quantitative RNA and protein gene expression for molecular-clinical correlations and how the RET pathway might be implicated in therapeutic success. Clinical data was collected during post-surgical follow-up. We also established new'humanized’ pituitary cultures, allowing 20 repeated passages and maintaining the pituitary secretory phenotype, and tested five multikinase inhibitors (TKI: Vandetanib, Lenvatinib, Sunitinib, Cabozantinib and Sorafenib) potentially able to act on the GDNF-induced RET dimerization/survival pathway. Antibody arrays investigated intracellular molecular pathways. Findings: In ACRO, there was specific enrichment of all genes in both RET pathways, especially GDNF. ARF and GFRA4 gene expression were found to be opposing predictors of response to first-line therapy. ARF cut-off levels, calculated categorizing by GNAS mutation, were predictive of good response (above) or resistance (below) to therapy months later. Sorafenib, through AMPK, blocked the GDNF/AKT survival action without altering the RET apoptotic pathway. Interpretation: Tumor ARF mRNA expression measured at the time of the surgery is a prognosis factor in acromegaly. The RET inhibitor, Sorafenib, is proposed as a potential treatment for resistant ACRO. Fund: This project was supported by national grants from Agencia Estatal de Investigación (AEI) and Instituto Investigación Carlos III, with participation of European FEDER funds, to IB (PI150056) and CVA (BFU2016-76973-R). It was also supported initially by a grant from the Investigator Initiated Research (IIR) Program (WI177773) and by a non-restricted Research Grant from Pfizer Foundation to IB. Some of the pituitary acromegaly samples were collected in the framework of the Spanish National Registry of Acromegaly (REMAH), partially supported by an unrestricted grant from Novartis to the Spanish Endocrine Association (SEEN). CVA is also supported from a grant of Medical Research Council UK MR/M018539/1

Forward SCT Module Assembly and Quality Control at IFIC Valencia

This note discusses the assembly and the quality control tests of 282 forward detector modules for the ATLAS Semiconductor Tracker assembled at the Instituto de Fisica Corpuscular (IFIC) in Valencia. The construction and testing procedures are outlined and the laboratory equipment is briefly described. Emphasis is given on the module quality achieved in terms of mechanical and electrical stability