Effects of Fetal Bovine Serum deprivation in cell cultures on the production of Anticarsia gemmatalis Multinucleopolyhedrovirus

<p>Abstract</p> <p>Background</p> <p><it>Anticarsia gemmatalis </it>is a pest in South America's soybean crops, which could be controlled by the Multinucleopolyhedrovirus of <it>A. gemmatalis </it>(AgMNPV). Currently, its commercial production is based on infected larvae. However, the possibility of using modified baculoviruses in Integrated Pest Management programs has stimulated an interest to develop alternative multiplication processes. This study evaluated the AgMNPV production in UFL-Ag-286 cells previously deprived Fetal Bovine Serum.</p> <p>Results</p> <p>Culture media containing 1% FBS during the previous 48 hours achieved a synchronized condition where 90% of cells were found in G₀/G₁stage, showing the presence of non-filamentous actin. All characteristics were estimated from cellular viability tests, cell actin detection trials and flow cytometer cell cycle analysis. AgMNPV production was tested by transcript studies and budded viruses (BVs) and occlusion bodies (OBs) yield quantitation. Results showed that the productivity in FBS deprived cells was 9.8 times more in BVs and 3.8 times more in OBs with respect to non-treated cells.</p> <p>Conclusions</p> <p>UFL-Ag-286 cells previously deprived in FBS shown to be a better host for AgMNPV propagation, increasing the useful for both <it>in vitro </it>bioinsecticide production and applications such as recombinant protein expression or gene delivery.</p

Assessment of aggressive bladder cancer mutations in plasma cell-free DNA

Background and aimsThe spatial and temporal genetic heterogeneity of bladder cancer (BC) makes challenging to find specific drivers of metastatic disease, thus preventing to determine those BC patients at high risk of tumor progression. Our aim was to identify DNA mutations providing aggressive behavior to bladder tumors and analyze them in patients’ cell-free DNA (cfDNA) during their follow-up after radical cystectomy (RC) in order to monitor tumor evolution.MethodsSix BC patients who underwent RC and presented disease progression during their follow-up were included. Next-generation sequencing was used to determine somatic mutations in several primary tumor and metastatic specimens from each patient. Shared DNA mutations between primary bladder tumor and metastatic sites were identified in cfDNA samples through droplet digital PCR.ResultsBesides BC genetic heterogeneity, specific mutations in at least one of these genes —TERT, ATM, RB1, and FGFR3— were found in primary tumors and their metastases in all patients. These mutations were also identified in the patients’ cfDNA at different follow-up time points. Additionally, the dynamic changes of these mutations in cfDNA allowed us to determine tumor evolution in response to treatment.ConclusionThe analysis of BC mutations associated with poor prognosis in plasma cfDNA could be a valuable tool to monitor tumor evolution, thus improving the clinical management of BC patients

Cell-Free DNA as a Prognostic Biomarker for Monitoring Muscle-Invasive Bladder Cancer

Cell-free DNA (cfDNA) has recently emerged as a real-time biomarker for diagnosis, monitoring and prediction of therapy response in tumoral disease. Here, we evaluated cfDNA as a prognostic biomarker for monitoring muscle-invasive bladder cancer (MIBC) patients at different follow-up time points. Blood samples from 37 MIBC patients who underwent radical cystectomy (RC) were collected at cystectomy and 1, 4, 12 and 24 months later. Plasma cfDNA amount and fragmentation patterns were determined. Four mutations were analyzed in cfDNA to detect circulating tumor DNA (ctDNA) during patient follow-up. During a median follow-up of 36 months, 46% of patients progressed; median time to progression was 10 months. cfDNA levels and ctDNA status four months after RC were identified as independent prognostic biomarkers of tumor progression (HR 5.290; p = 0.033) and cancer-specific survival (HR 4.199; p = 0.038), respectively. Furthermore, ctDNA clearance four months after RC was significantly associated with patients’ clinical outcomes. In conclusion, cfDNA levels and ctDNA status four months after RC have prognostic implications in MIBC patients. In addition, cfDNA monitoring is useful to predict patient outcomes after RC. cfDNA analysis in the clinical setting could greatly improve MIBC patient management.This research was funded by the Instituto de Salud Carlos III (ISCIII) through the Plan Estatal de Investigación Científica y Técnica y de Innovación 2018–2020, project reference number PI17/01343, and co-funded by the European Regional Development Fund (ERDF)

Prognostic Gene Expression-Based Signature in Clear-Cell Renal Cell Carcinoma

The inaccuracy of the current prognostic algorithms and the potential changes in the therapeutic management of localized ccRCC demands the development of an improved prognostic model for these patients. To this end, we analyzed whole-transcriptome profiling of 26 tissue samples from progressive and non-progressive ccRCCs using Illumina Hi-seq 4000. Differentially expressed genes (DEG) were intersected with the RNA-sequencing data from the TCGA. The overlapping genes were used for further analysis. A total of 132 genes were found to be prognosis-related genes. LASSO regression enabled the development of the best prognostic six-gene panel. Cox regression analyses were performed to identify independent clinical prognostic parameters to construct a combined nomogram which includes the expression of CERCAM, MIA2, HS6ST2, ONECUT2, SOX12, TMEM132A, pT stage, tumor size and ISUP grade. A risk score generated using this model effectively stratified patients at higher risk of disease progression (HR 10.79; p < 0.001) and cancer-specific death (HR 19.27; p < 0.001). It correlated with the clinicopathological variables, enabling us to discriminate a subset of patients at higher risk of progression within the Stage, Size, Grade and Necrosis score (SSIGN) risk groups, pT and ISUP grade. In summary, a gene expression-based prognostic signature was successfully developed providing a more precise assessment of the individual risk of progression

Combining SfM Photogrammetry and Terrestrial Laser Scanning to Assess Event-Scale Sediment Budgets along a Gravel-Bed Ephemeral Stream

[EN] Stream power represents the rate of energy expenditure along a stream reach and can be calculated using topographic data acquired via structure-from-motion (SfM) photogrammetry and terrestrial laser scanning (TLS). This study sought to quantitatively relate morphological adjustments in the Azohia Rambla, a gravel-bed ephemeral stream in southeastern Spain, to stream power (omega), critical power (omega(c)), and energy gradients ( partial differential omega/ partial differential s), along different reference channel reaches of 200 to 300 m in length. High-resolution digital terrain models (HRDTMs), combined with ortophotographs and point clouds from 2018, 2019, and 2020, and ground-based surveys, were used to estimate the spatial variability of morphological sediment budgets and to assess channel bed mobility during the study period at different spatial scales: reference channel reaches (RCRs), pilot bed survey areas (PBSAs), and representative geomorphic units (RGUs). The optimized complementary role of the SfM technique and terrestrial laser scanning allowed the generation of accurate and reliable HRDTMs, upon which a 1-D hydrodynamic model was calibrated and sediment budgets calculated. The resulting high-resolution maps allowed a spatially explicit analysis of stream power and transport efficiency in relation to volumes of erosion and deposition in the RCR and PBSA. In addition, net incision or downcutting and vertical sedimentary accretion were monitored for each flood event in relation to bedforms and hydraulic variables. Sediment sources and sinks and bed armoring processes showed different trends according to the critical energy and stream power gradient, which were verified from field observations. During flows exceeding bankfull discharges (between 18 and 24 m(3) s(-1) according to channel reach), significant variations in partial differential omega/ partial differential s values and omega/omega(c) ratios (e.g., -15 2 for a peak discharge of 31 m(3) s(-1)) were associated with a large amount of bedload mobilized upstream and vertical accretion along the middle reach (average rise height of 0.20 to 0.35 m for the same event). By contrast, more moderate peak flows (<= 10 m(3) s(-1)) only produced minor changes resulting in surface washing, selective transport, and local bed scouring.This research was funded by ERDF/Spanish Ministry of Science, Innovation and Universities-State Research Agency/Project CGL2017-84625-C2-1-R (CCAMICEM); State Program for Research, Development and Innovation Focused on the Challenges of Society, the Spanish Ministry of Economy and Competitiveness (MINECO) and EU FEDER under Project TEC2017-85244-C2-1-P and by the University of Alicante (vigrob-157 and GRE18-05).Conesa-García, C.; Puig-Mengual, C.; Riquelme, A.; Tomás, R.; Martinez-Capel, F.; García-Lorenzo, R.; Pastor, JL.... (2020). Combining SfM Photogrammetry and Terrestrial Laser Scanning to Assess Event-Scale Sediment Budgets along a Gravel-Bed Ephemeral Stream. Remote Sensing. 12(21):1-27. https://doi.org/10.3390/rs12213624S127122

La evaluación del abuso sexual en personas con discapacidad intelectual: proceso de construcción y validación de un instrumento de autoinforme

People with functional diversity have certain limitations in the functioning of their daily life, which makes them dependent on their primary caregivers, leaving their bodies exposed to them. In addition, they have fewer resources to detect and defend themselves against abuse. These factors, among others, make them a group vulnerable to sexual abuse. But in the detection of potential victims, we find limitations in comprehension and literacy, which makes questioning the reliability of the information obtained. The objective of this study is the development of a reliable and valid instrument adapted to the characteristics of people with functional diversity and allows the detection of cases of sexual abuse. In order to do so, the construction-validation process of the scale has been carried out in two phases, generating, in each of them, different versions that have been applied to two groups of participants. It details the analysis that the group of experts has made from the two pilot applications and the consequent modifications derived from these analyzes. In the first phase the version applies to 20 participants and in the second to 150. The final version has a reliability of .91. The study has shown the complexity of developing an instrument for assessing sexual abuse for people with intellectual disabilities because of the limitations of understanding inherent in the collective, stressing the importance of having the contributions of professionals who work daily with them.Las personas con diversidad funcional presentan ciertas limitaciones en el funcionamiento desu día a día, lo que les convierten en dependientes de sus cuidadores principales, quedando su cuerpo expuesto a ellos. Además, presentan menos recursos para detectar y defenderse ante situaciones de abuso. Estos factores, entre otros, hacen que se conviertan en un colectivo vulnerable al abuso sexual. Pero en la detección de las víctimas potenciales, nos encontramos con limitaciones en comprensión y lectoescritura, lo que hace cuestionar la fiabilidad de la información obtenida. El objetivo de este estudio, es la elaboración de un instrumento fiable y válido que se adapte a las características de las personas con diversidad funcional, y posibilite la detección de casos de abuso sexual. Para ello, el proceso de construcción-validación de la escala se ha realizado en dos fases generándose, en cada una de ellas, diferentes versiones que se han aplicado a dos grupos de participantes.Se detalla el análisis que el grupo de expertos ha realizado a partir de las dos aplicacionespiloto y las consecuentes modificaciones derivadas de dichos análisis. En la primera fase la versión se aplica a 20 participantes y en la segunda a 150. La versión final cuenta con una fiabilidad de .91.El estudio ha mostrado la complejidad que supone elaborar un instrumento de evaluación del abuso sexual para personas con discapacidad intelectual por las limitaciones de comprensión inherentes al colectivo, remarcando la importancia de contar con las aportaciones de profesionales que trabajan diariamente con ellos

Cell-Free DNA as a Prognostic Biomarker for Monitoring Muscle-Invasive Bladder Cancer

Cell-free DNA (cfDNA) has recently emerged as a real-time biomarker for diagnosis, monitoring and prediction of therapy response in tumoral disease. Here, we evaluated cfDNA as a prognostic biomarker for monitoring muscle-invasive bladder cancer (MIBC) patients at different follow-up time points. Blood samples from 37 MIBC patients who underwent radical cystectomy (RC) were collected at cystectomy and 1, 4, 12 and 24 months later. Plasma cfDNA amount and fragmentation patterns were determined. Four mutations were analyzed in cfDNA to detect circulating tumor DNA (ctDNA) during patient follow-up. During a median follow-up of 36 months, 46% of patients progressed; median time to progression was 10 months. cfDNA levels and ctDNA status four months after RC were identified as independent prognostic biomarkers of tumor progression (HR 5.290; p = 0.033) and cancer-specific survival (HR 4.199; p = 0.038), respectively. Furthermore, ctDNA clearance four months after RC was significantly associated with patients' clinical outcomes. In conclusion, cfDNA levels and ctDNA status four months after RC have prognostic implications in MIBC patients. In addition, cfDNA monitoring is useful to predict patient outcomes after RC. cfDNA analysis in the clinical setting could greatly improve MIBC patient management

Differential gene expression profile between progressive and de novo muscle invasive bladder cancer and its prognostic implication

This study aimed to ascertain gene expression profle diferences between progressive muscle-invasive bladder cancer (MIBC) and de novo MIBC, and to identify prognostic biomarkers to improve patients' treatment. Retrospective multicenter study in which 212 MIBC patients who underwent radical cystectomy between 2000 and 2019 were included. Gene expression profles were determined in 26 samples using Illumina microarrays. The expression levels of 94 genes were studied by quantitative PCR in an independent set of 186 MIBC patients. In a median follow-up of 16 months, 46.7% patients developed tumor progression after cystectomy. In our series, progressive MIBC patients show a worse tumor progression (p= 0.024) and cancer-specifc survival (CSS) (p= 0.049) than the de novo group. A total of 480 genes were found to be diferently expressed between both groups. Diferential expression of 24 out of the 94 selected genes was found in an independent cohort. RBPMC2 and DSC3 were found as independent prognostic biomarkers of tumor progression and CALD1 and LCOR were identifed as prognostic biomarkers of CSS between both groups. In conclusion, progressive and de novo MIBC patients show diferent clinical outcome and gene expression profles. Gene expression patterns may contribute to predict high-risk of progression to distant metastasis or CSS