Combined flow cytometry and high-throughput image analysis for the study of essential genes in Caenorhabditis elegans

Background: Advances in automated image-based microscopy platforms coupled with high-throughput liquid workflows have facilitated the design of large-scale screens utilising multicellular model organisms such as Caenorhabditis elegans to identify genetic interactions, therapeutic drugs or disease modifiers. However, the analysis of essential genes has lagged behind because lethal or sterile mutations pose a bottleneck for high-throughput approaches, and a systematic way to analyse genetic interactions of essential genes in multicellular organisms has been lacking. Results: In C. elegans, non-conditional lethal mutations can be maintained in heterozygosity using chromosome balancers, commonly expressing green fluorescent protein (GFP) in the pharynx. However, gene expression or function is typically monitored by the use of fluorescent reporters marked with the same fluorophore, presenting a challenge to sort worm populations of interest, particularly at early larval stages. Here, we develop a sorting strategy capable of selecting homozygous mutants carrying a GFP stress reporter from GFP-balanced animals at the second larval stage. Because sorting is not completely error-free, we develop an automated high-throughput image analysis protocol that identifies and discards animals carrying the chromosome balancer. We demonstrate the experimental usefulness of combining sorting of homozygous lethal mutants and automated image analysis in a functional genomic RNA interference (RNAi) screen for genes that genetically interact with mitochondrial prohibitin (PHB). Lack of PHB results in embryonic lethality, while homozygous PHB deletion mutants develop into sterile adults due to maternal contribution and strongly induce the mitochondrial unfolded protein response (UPR mt ). In a chromosome-wide RNAi screen for C. elegans genes having human orthologues, we uncover both known and new PHB genetic interactors affecting the UPR mt and growth. Conclusions: The method presented here allows the study of balanced lethal mutations in a high-throughput manner. It can be easily adapted depending on the user's requirements and should serve as a useful resource for the C. elegans community for probing new biological aspects of essential nematode genes as well as the generation of more comprehensive genetic networks.European Research Council ERC-2011-StG-281691Ministerio de Economía y Competitividad BFU2012–3550

Segmentation of Magnetic Resonance Imaging MRI using LS-SVM and Wavelet Multiresolution Analysis

Actualmente, las máquinas de soporte vectorial (SVM) se han convertido en una herramienta poderosa para resolver problemas de clasificación no lineal. Para la optimización de esta herramienta, se ha desarrollado una reformulación conocida como LS-SVM (máquina de soporte vectorial de mínimos cuadrados), la cual trabaja con un modelo de minimización basada en funciones y polinomios de Lagrange. Por lo tanto, este trabajo presenta un método para la segmentación de imágenes de resonancia magnética específicamente para estudiar la morfología de los pulmones y lograr la cuantificación de características relevantes en dichas imágenes usando SVM y LS-SVM. Adicionalmente a la técnica de clasificación, en este trabajo se usaron técnicas como: análisis Wavelet para eliminación de información no relevante (compresión), y algoritmos Splines, para interpolar la información encontrada y cuantificar las características, que se basaron en el reconocimiento de área, forma y estructuras anormales presentes en la zona pulmonar de dichas imágenes.Currently, support vector machines (SVM) have become a powerful tool to solve nonlinear classification problems. For the optimization of the tool, has developed a reformulation known as LS-SVM (Support Vector Machine least squares), which works with a model based on function minimization and Lagrange polynomials. Therefore, this paper presents a method for segmentation of magnetic resonance images specifically to study the morphology of the lungs and reach the quantification of relevant features in these images using SVM and LS-SVM. In addition to sorting technique in this work using techniques such as wavelet analysis to eliminate irrelevant information (compression) and Splines algorithms to interpolate the information found and quantify the characteristics, which in this work were based on the recognition area, shape and abnormal structures present in the lung of these images

Las asociaciones de ostrácodos en secuencias aluviales como indicadores de cambios ambientales holocenos (Bardenas Reales de Navarra, Cuenca del Ebro, NE Península Ibérica)

En este trabajo se estudian las asociaciones de ostrácodos identificadas en tres unidades aluviales preservadas en Bardenas Reales de Navarra (Cuenca del Ebro, NE Península Ibérica), para reconstruir la evolución paleoambiental de esta área durante el Holoceno final. Las especies de ostrácodos más comunes en las muestras estudiadas son Ilyocypris bradyi Sars y Paralimnocythere aff. psammophila (Flössner), acompañadas por Pseudocandona albicans (Brady). Las variaciones en su abundancia relativa a lo largo de dichas unidades aluviales permiten caracterizar cambios internos en el ambiente de depósito. Así, la unidad depositada entre 4763±87 y 2848±55 años cal. BP presenta como especie más abundante a I. bradyi, lo que indica la existencia de pequeños arroyos o riachuelos con agua corriente que se desarrollaron probablemente en llanuras de inundación fluviales. Sin embargo, de 2848±55 a 836±65 años cal. BP Paralimnocythere aff. psammophila es la especie dominante, representando el establecimiento de un medio acuático efímero y de aguas estancadas, posiblemente relacionado con condiciones climáticas más áridas (Anomalía Climática Medieval). Finalmente, la asociación de ostrácodos identificada en el Reciente (191±97 a 127±82 años cal. BP) está constituida principalmente por I. bradyi, que indicaría la presencia de nuevo de aguas corrientes. Esta última asociación parece estar vinculada a la influencia de condiciones climáticas húmedas y frías (Pequeña Edad del Hielo) durante el Reciente en esta área. Por lo tanto, la evolución paleoambiental para el Holoceno final de Bardenas Reales de Navarra, identificada a partir de las asociaciones de ostrácodos, muestra una buena correlación con los cambios climáticos rápidos holocenos deducidos en otros registros continentales del NE de la Península Ibérica. In this paper, the ostracod assemblages identified in three alluvial units preserved in Bardenas Reales de Navarra (Ebro Basin, NE Iberian Peninsula) have been studied, with the aim of reconstructing the palaeoenvironmental evolution of this area during the late Holocene. The most common ostracod species in the studied samples are Ilyocypris bradyi Sars and Paralimnocythere aff. psammophila (Flössner), accompanied by Pseudocandona albicans (Brady). Variations in their relative abundance along these alluvial units allow us to characterize internal changes in the depositional environment. Thus, the unit deposited between 4763±87 and 2848±55 years cal. BP is dominated by I. bradyi, indicating the presence of small streams with running water developed probably in fluvial flood plains. Nevertheless, from 2848±55 to 836±65 years cal. BP Paralimnocythere aff. psammophila is the dominant species, representing the establishment of an ephemeral and stagnant aquatic environment, mainly related with more arid climatic conditions (Medieval Warm Period). Finally, the ostracod assemblage identified in Recent times (191±97 to 127±82 years cal. BP) is mainly dominated by I. bradyi, which indicates the presence of running waters. This later assemblage probably defines the influence of cold and wet climatic phase (Little Ice Age) during Recent times in this area. The palaeoenvironmental evolution identified from the late Holocene according to ostracod assemblages in Bardenas Reales de Navarra shows good correlation with rapid climatic changes deduced from other continental records of the NE Iberian Peninsula

Magnetic susceptibility studies of the spin-glass and Verwey transitions in magnetite nanoparticles

Magnetite nanostructured powder samples were synthesized by aging chemical method. Phase, structural, and magnetic properties were characterized. X-ray diffraction patterns showed cubic magnetite pure phase, with average crystallite size, , equal to 40 nm. Susceptibility measurements showed the well-known Verwey transition at a temperature of 90 K. The decrease of Verwey transition temperature, with respect to the one reported in literature (125 K) was attributed to the low average crystallite size. Moreover, the spin-glass like transition was observed at 35 K. Activation energy calculated from susceptibility curves, with values ranging from 6.26 to 6.93 meV, showed a dependence of spin-glass transition on frequency. Finally, hysteresis loops showed that there is not an effect of Verwey transition on magnetic properties. On the other hand, a large increase of coercivity and remanent magnetization at a temperature between 5 and 50 K confirmed the presence of a magnetic transition at low temperatures

Síndrome neuroléptico maligno inducido por risperidona y facilitado por sepsis de origen urinario: clínica y fisiopatología.

Risperidone is a selective monoaminergic antagonist with a high affinity for dopamine receptors that can cause neuroleptic malignant syndrome (NMS), considered a life-threatening medical emergency. It has an incidence of up to 3% and its mortality is between 10 and 20%. The case of a 56-year-old female who met the clinical criteria of NMS, induced by the use of risperidone and facilitated by a sepsis of urinary origin, is reported. It was managed with a dopamine agonist and the change of antipsychotic, which resulted in a favorable clinical course. The NMS is a low-prevalence entity whose diagnosis has specificity and sensitivity greater than 90%, reason for which must be clearly differentiated from other pathologies. The mechanism by which urinary infection could facilitate the occurrence of this disease is discussed. Early diagnosis improves the response to an adequate management to be established in each case.La risperidona es un antagonista selectivo monoaminérgico, con una elevada afinidad por receptores dopaminérgicos, que puede producir síndrome neuroléptico maligno (SNM), considerado una emergencia médica con alto riesgo de muerte. Tiene una incidencia de hasta el 3% y su mortalidad está entre el 10 y 20%. Se reporta el caso de una paciente de 56 años, que reunía los criterios clínicos del SNM, inducido por el uso de risperidona y facilitado por una sepsis de origen urinario. El tratamiento se condujo con un agente agonista dopaminérgico y cambio del antipsicótico, procedimientos que resultaron en una adecuada evolución clínica. El SNM es una entidad de baja prevalencia, para la cual existen criterios diagnósticos con especificidad y sensibilidad mayor del 90%, por lo que debe diferenciarse claramente de otras patologías. Se discute el mecanismo mediante el cual la infección urinaria facilitaría la ocurrencia de esta enfermedad. El diagnóstico precoz mejora la respuesta al manejo adecuado que se establezca en cada caso

Magnetic phase diagram of nanostructured zinc ferrite as a function of inversion degree delta

Magnetic properties of spinel zinc ferrites are strongly linked to the synthesis method and the processing route since they control the microstructure of the resulting material. In this work, ZnFe_2O_4 nanoparticles were synthesized by the mechanochemical reaction of stoichiometric ZnO and alpha-Fe2O3, and single-phase ZnFe_2O_4 was obtained after 150 h of milling. The as-milled samples, with a high inversion degree, were subjected to different thermal annealings up to 600 ºC to control the inversion degree and, consequently, the magnetic properties. The as-milled samples, with a crystallite size of 11 nm and inversion degree delta = 0.57, showed ferrimagnetic behavior even above room temperature, as shown by Rietveld refinements of the X-ray diffraction pattern and superconducting quantum interference device magnetometry. The successive thermal treatments at 300, 400, 500, and 600 degrees C decrease delta from 0.15 to 0.18, affecting the magnetic properties. A magnetic phase diagram as a function of delta can be inferred from the results: for delta 0.5, a new antiferromagnetic order appeared due to the overpopulation of nonmagnetic Zn on octahedral sites that leads to equally distributed magnetic cations in octahedral and tetrahedral sites

Management of incidentally discovered appendiceal neuroendocrine tumors after an appendicectomy

Appendiceal neoplasms; Carcinoid tumor; Treatment outcomeNeoplasias apendiculares; Tumor carcinoide; Resultado del tratamientoNeoplàsies apendiculars; Tumor carcinoide; Resultat del tractamentAppendiceal neuroendocrine tumors (aNETs) are an uncommon neoplasm that is relatively indolent in most cases. They are typically diagnosed in younger patients than other neuroendocrine tumors and are often an incidental finding after an appendectomy. Although there are numerous clinical practice guidelines on management of aNETs, there is continues to be a dearth of evidence on optimal treatment. Management of these tumors is stratified according to risk of locoregional and distant metastasis. However, there is a lack of consensus regarding tumors that measure 1-2 cm. In these cases, some histopathological features such as size, tumor grade, presence of lymphovascular invasion, or mesoappendix infiltration must also be considered. Computed tomography or magnetic resonance imaging scans are recommended for evaluating the presence of additional disease, except in the case of tumors smaller than 1 cm without additional risk factors. Somatostatin receptor scintigraphy or positron emission tomography with computed tomography should be considered in cases with suspected residual or distant disease. The main point of controversy is the indication for performing a completion right hemicolectomy after an initial appendectomy, based on the risk of lymph node metastases. The main factor considered is tumor size and 2 cm is the most common threshold for indicating a colectomy. Other factors such as mesoappendix infiltration, lymphovascular invasion, or tumor grade may also be considered. On the other hand, potential complications, and decreased quality of life after a hemicolectomy as well as the lack of evidence on benefits in terms of survival must be taken into consideration. In this review, we present data regarding the current indications, outcomes, and benefits of a colectomy

Randomized phase II trial of FOLFIRI-panitumumab compared with FOLFIRI alone in patients with RAS wild-type circulating tumor DNA metastatic colorectal cancer beyond progression to first-line FOLFOX-panitumumab : the BEYOND study (GEMCAD 17-01)

Altres ajuts: Fundació la Marató de TV3 (201330.10); Fundacion Olga Torres (Modalitat A. 2019/2020); Spanish Association Against Cancer (AECC, PROYE19040POST_001).Purpose: Panitumumab plus FOLFOX (P-FOLFOX) is standard first-line treatment for RAS wild-type (WT) metastatic colorectal cancer. The value of panitumumab rechallenge is currently unknown. We assessed addition of panitumumab to FOLFIRI (P-FOLFIRI) beyond progression to P-FOLFOX in patients with no RAS mutations in liquid biopsy (LB). Methods: In this randomized phase II trial, patients were assigned (3:2 ratio) to second-line P-FOLFIRI (arm A) or FOLFIRI alone (arm B). LB for circulating tumor DNA analysis was collected at study entry and at disease progression. Primary endpoint was 6-month progression-free survival. Two-stage Simon design required 85 patients to be included (EudraCT 2017-004519-38). Results: Between February 2019 and November 2020, 49 patients were screened (16 RAS mutations in LB detected) and 31 included (18 assigned to arm A and 13 to arm B). The study was prematurely closed due to inadequate recruitment. Serious adverse events were more frequent in arm A (44% vs. 23%). Overall response rate was 33% (arm A) vs. 7.7% (arm B). Six-month progression-free survival rate was 66.7% (arm A) and 38.5% (arm B). Median progression-free survival was 11.0 months (arm A) and 4.0 months (arm B) (hazard ratio, 0.58). At disease progression, RAS or BRAF mutations in LB were found in 4/11 patients (36%) in arm A and 2/10 (20%) in arm B. Conclusions: The BEYOND study suggests a meaningful benefit of P-FOLFIRI beyond progression to P-FOLFOX in metastatic colorectal cancer patients with WT RAS status selected by LB. This strategy deserves further investigation