Implicación de la proteína clusterina y la actividad apoptótica en la transformación maligna y el pronóstico de los pacientes con cáncer colorrectal

Introducción: La carcinogénesis colorrectal es un proceso de múltiples etapas que progresa desde el tejido normal a adenoma y finalmente carcinoma. Se ha relacionado a la clusterina (CLU), una glicoproteína heterodimérica ampliamente distribuida en el organismo de los mamíferos, con la supervivencia celular, la respuesta al estrés y la resistencia a los tratamientos antineoplásicos. De hecho, existen terapias anti-CLU con oliglonucleótidos antisentido capaces de potenciar la apoptosis inducida por los citotóxicos convencionales. Sin embargo, la evidencia disponible sobre la participación de CLU en la progresión tumoral del colon es limitada y poco sabemos acerca de su valor pronóstico en el cáncer colorrectal (CCR). Además, el papel que juega la apoptosis en esta secuencia de carcinogénesis colorrectal no ha sido completamente clarificado, y los trabajos que han investigado la relación entre la actividad apoptótica y la supervivencia de los pacientes con CCR han arrojado resultados, hasta la fecha, contradictorios. Objetivos: Los objetivos del estudio fueron estudiar la relación de CLU con la carcinogénesis y la progresión tumoral del CCR, así como su significado pronóstico, analizando el impacto de la expresión de CLU en la supervivencia de los pacientes con CCR. Asimismo, se quiso determinar la influencia de la actividad apoptótica en la carcinogénesis y el pronóstico del CCR. Material y Métodos: Examinamos las muestras de 103 CCRs resecados en el Hospital Costa del Sol, 31 adenomas y 20 epitelios normales. CLU se determinó mediante inmunohistoquímica, usando un anticuerpo monoclonal anti-cadena α (Upstate-Millipore, Watford, Inglaterra). La expresión de CLU fue considerada negativa (CLU-) si no se observaba tinción y positiva (CLU+) cuando >10% de células resultaban teñidas. Para la detección de la apoptosis. se llevó a cabo el ensayo TUNEL. La media de porcentaje apoptótico (1%, rango 0-6%) fue el punto de corte empleado para los estudios de supervivencia. La estimación de la supervivencia se hizo mediante el método de Kaplan-Meier y usamos la regresión de Cox para el análisis multivariante (nivel de significación estadística p<0.05). La mediana de seguimiento fue 54 meses. La mediana de edad fue 70 años (rango 45-91) y la distribución por estadios: I (15%), II (48%), III (23%) y IV (14%). Resultados: Las células epiteliales del tejido normal fueron CLU-. En cambio el 16% (5/31) de los adenomas fue CLU+ y este porcentaje se incrementó en los CCRs (30%, 31/103). Las recidivas fueron más frecuentes en los tumores CLU+ (61%,19/31) que en los CLU- (37%, 27/72) y la expresión de CLU se asoció significativamente con una menor supervivencia libre de progresión (SLP) (SLP a 5 años: 35 ± 9% vs 57 ± 7%; p 0.03). En el análisis multivariante, CLU (HR=2.01, IC 95% 1.03-3.90) y el estadio TNM (HR=2.20; IC 95% 1.13-4.29) fueron factores pronósticos independentes para la SLP. Encontramos un índice apoptótico (IA) mayor en los CCRs (1.09 ± 0.13) que en los adenomas (0.38 ± 0.23, p=0.059) y significativamente mayor que en el epitelio normal (0.06 ± 0.04, p=0.001). El estadio IV mostró un IA superior en comparación con otros estadios (p=0.017). El IA alto se asoció con una menor SLP (SLP a 5 años: 28.5 ± 9.6% vs 61.8 ± 8.1%; p<0.05) y supervivencia global (SG) (SG a 5 años: 43.7 ± 11.9% vs 74.6 ± 8.8%; p <0.05). En el análisis multivariante, el IA (HR=2.18, IC 95% 1.08-4.37) y el estadio TNM (HR=2.41, 95% CI 1.20-4.85) resultaron factores pronósticos independientes para la SG. Conclusiones: Tanto CLU como la actividad apoptótica aumentan progresivamente a lo largo de la secuencia mucosa sana-adenoma-carcinoma, lo que sugiere un papel relevante en la carcinogénesis colorrectal. La sobreexpresión de CLU se asocia con un peor pronóstico de los pacientes con CCR, por lo que nos permite identificar a pacientes con tumores de comportamiento más agresivo. Un IA elevado se asocia también con una mayor agresividad tumoral y una peor supervivencia del CCR

Testing PtCu nanoparticles supported on highly ordered mesoporous carbons CMK3 and CMK8 as catalysts for low-temperature fuel cells

Pt(Cu) nanoparticles supported on CMK3 and CMK8 ordered mesoporous carbons (OMCs) have been synthesized by electroless deposition of Cu followed by galvanic exchange with Pt. The structural characterization by high-resolution transmission electron microscopy and X-ray diffraction showed the formation of Pt(Cu) nanoparticles of 4-5 nm, in which PtCu alloys with contracted fcc Pt lattice and 70-80 at.% Pt was identified. The X-ray photoelectron spectroscopy analyses indicated that the Pt(Cu) nanoparticles were mainly composed of a PtCu alloy core covered by a Ptrich shell, in agreement with the steady cyclic voltammograms, which did not show any Cu oxidation peaks. Electroactive surface areas up to about 70 m2 gPt−1 were obtained. The onset potentials for CO oxidation and the oxygen reduction reaction were more negative and positive, respectively, as compared to Pt/C, thus indicating higher activity of these Pt(Cu) catalysts with respect to the latter. Based on the corresponding binding energies, these better activities were attributed to the favorable geometric and ligand effects of Cu on Pt, which were able to reduce the adsorption energy of the intermediates on Pt. Pt(Cu)/CMK3 showed competitive mass and specific activities, as well as better stability than Pt/C

Electrochemical performance of carbon-supported Pt(Cu) electrocatalysts for low-temperature fuel cells

Pt(Cu) nanoparticles supported on carbon nanofibers (CNFs), multi-walled carbon nanotubes (MWCNTs) and Vulcan carbon XC72, have been synthesized by electroless deposition and galvanic exchange. The structural analyses show contracted Pt fcc lattices due to the formation of a PtCu alloy core covered by a Pt-rich shell, mean crystallite sizes of about 3 nm, as well as good dispersion and carbon attachment. The electrochemical surface areas (ECSAs) of Pt(Cu)/CNF and Pt(Cu)/XC72 are comparable to those of commercial Pt/C and PtCu/C. The Pt(Cu) electrocatalysts show more negative onset potentials for CO oxidation than Pt/C and PtCu/C, thus indicating their greater CO tolerance. Pt(Cu)/CNF and Pt(Cu)/MWCNT present the highest mass activity and specific activity for the O2 reduction, respectively, both with better relative stability than Pt(Cu)/XC72. Pt(Cu)/CNF and Pt(Cu)/MWCNT are then considered good cathode catalysts, yielding estimated savings of about 50 wt.% Pt, when applied to low-temperature fuel cells

8-oxoguanine DNA glycosylase 1 upregulation as a risk factor for obesity and colorectal cancer

DNA damage has been extensively studied as a potentially helpful tool in assessing and preventing cancer, having been widely associated with the deregulation of DNA damage repair (DDR) genes and with an increased risk of cancer. Adipose tissue and tumoral cells engage in a reciprocal interaction to establish an inflammatory microenvironment that enhances cancer growth by modifying epigenetic and gene expression patterns. Here, we hypothesize that 8-oxoguanine DNA glycosylase 1 (OGG1)—a DNA repair enzyme—may represent an attractive target that connects colorectal cancer (CRC) and obesity. In order to understand the mechanisms underlying the development of CRC and obesity, the expression and methylation of DDR genes were analyzed in visceral adipose tissue from CRC and healthy participants. Gene expression analysis revealed an upregulation of OGG1 expression in CRC participants (p < 0.005) and a downregulation of OGG1 in normal-weight healthy patients (p < 0.05). Interestingly, the methylation analysis showed the hypermethylation of OGG1 in CRC patients (p < 0.05). Moreover, expression patterns of OGG1 were found to be regulated by vitamin D and inflammatory genes. In general, our results showed evidence that OGG1 can regulate CRC risk through obesity and may act as a biomarker for CRC.Partial funding for open access charge: Universidad de Málag

Alterations of perineuronal nets in the dorsolateral prefrontal cortex of neuropsychiatric patients

Background: Alterations in the structure and physiology of interneurons in the prefrontal cortex (PFC) are important factors in the etiopathology of different psychiatric disorders. Among the interneuronal subpopulations, parvalbumin (PV) expressing cells appear to be specially affected. Interestingly, during development and adulthood the connectivity of these interneurons is regulated by the presence of perineuronal nets (PNNs), specialized regions of the extracellular matrix, which are frequently surrounding PV expressing neurons. Previous reports have found anomalies in the density of PNNs in the PFC of schizophrenic patients. However, although some studies have described alterations in PNNs in some extracortical regions of bipolar disorder patients, there are no studies focusing on the prefrontocortical PNNs of bipolar or major depression patients. For this reason, we have analyzed the density of PNNs in post-mortem sections of the dorsolateral PFC (DLPFC) from the Stanley Neuropathology Consortium, which includes controls, schizophrenia, bipolar and major depression patients. Results: We have not observed differences in the distribution of PV+ cells or PNNs, or in the percentage of PV+ interneurons surrounded by PNNs. The density of PV+ interneurons was similar in all the experimental groups, but there was a significantly lower density of PNNs in the DLPFC of bipolar disorder patients and a tendency towards a decrease in schizophrenic patients. No differences were found when evaluating the density of PV+ cells surrounded by PNNs. Interestingly, when assessing the influence of demographic data, we found an inverse correlation between the density of PNNs and the presence of psychosis. Conclusions: The present results point to prefrontocortical PNNs and their role in the regulation of neuronal plasticity as putative players in the etiopathology of bipolar disorder and schizophrenia. Our findings also suggest a link between these specialized regions of the extracellular matrix and the presence of psychosis

Detection of Minority Variants and Mixed Infections in Mycobacterium tuberculosis by Direct Whole-Genome Sequencing on Noncultured Specimens Using a Specific-DNA Capture Strategy

Detection of mixed Mycobacterium tuberculosis (MTB) infections is essential, particularly when resistance mutations are present in minority bacterial populations that may affect patients' disease evolution and treatment. Whole-genome sequencing (WGS) has extended the amount of key information available for the diagnosis of MTB infection, including the identification of mixed infections. Having genomic information at diagnosis for early intervention requires carrying out WGS directly on the clinical samples. However, few studies have been successful with this approach due to the low representation of MTB DNA in sputa. In this study, we evaluated the ability of a strategy based on specific MTB DNA enrichment by using a newly designed capture platform (MycoCap) to detect minority variants and mixed infections by WGS on controlled mixtures of MTB DNAs in a simulated sputum genetic background. A pilot study was carried out with 12 samples containing 98% of a DNA pool from sputa of patients without MTB infection and 2% of MTB DNA mixtures at different proportions. Our strategy allowed us to generate sequences with a quality equivalent to those obtained from culture: 62.5× depth coverage and 95% breadth coverage (for at least 20× reads). Assessment of minority variant detection was carried out by manual analysis and allowed us to identify heterozygous positions up to a 95:5 ratio. The strategy also automatically distinguished mixed infections up to a 90:10 proportion. Our strategy efficiently captures MTB DNA in a nonspecific genetic background, allows detection of minority variants and mixed infections, and is a promising tool for performing WGS directly on clinical samples. IMPORTANCE We present a new strategy to identify mixed infections and minority variants in Mycobacterium tuberculosis by whole-genome sequencing. The objective of the strategy is the direct detection in patient sputum; in this way, minority populations of resistant strains can be identified at the time of diagnosis, facilitating identification of the most appropriate treatment for the patient from the first moment. For this, a platform for capturing M. tuberculosis-specific DNA was designed to enrich the clinical sample and obtain quality sequences

Pembrolizumab in Asian patients with microsatellite-instability-high/mismatch-repair-deficient colorectal cancer

The phase 3 KEYNOTE-177 study evaluated pembrolizumab versus chemotherapy with or without bevacizumab or cetuximab in patients with newly diagnosed, microsatellite-instability-high (MSI-H)/mismatch-repair-deficient (dMMR) metastatic colorectal cancer (mCRC). Primary endpoints were progression-free survival (PFS) per RECIST v1.1 by blinded independent central review (BICR) and overall survival (OS). Secondary endpoints were overall response rate (ORR) per RECIST v1.1 by BICR and safety. Here, we report results from the post hoc analysis of patients who were enrolled in Asia from the final analysis (FA) of KEYNOTE-177. A total of 48 patients from Japan, Korea, Singapore, and Taiwan (pembrolizumab, n =?22; chemotherapy, n =?26) were included. At FA, median time from randomization to data cutoff (February 19, 2021) was 45.3 (range 38.1?57.8) months with pembrolizumab and 43.9 (range 36.6?55.1) months with chemotherapy. Median PFS was not reached (NR; 95% confidence interval [CI] 1.9 months?NR) with pembrolizumab versus 10.4 (95% CI 6.3?22.0) months with chemotherapy (hazard ratio [HR] 0.56, 95% CI 0.26?1.20). Median OS was NR (range 13.8 months?NR) versus 30.0 (14.7?NR) months (HR 0.65, 95% CI 0.27?1.55) and ORR was 50% (95% CI 28?72) versus 46% (95% CI 27?67). Grade 3/4 treatment-related adverse events (TRAEs) were reported by two patients (9%) in the pembrolizumab arm and 20 (80%) in the chemotherapy arm. Immune-mediated adverse events or infusion reactions were reported by six patients (27%) and 10 patients (40%), respectively. No deaths due to TRAEs occurred. These data support first-line pembrolizumab as a standard of care for patients from Asia with MSI-H/dMMR mCRC. ClinicalTrials.gov identifier: NCT02563002.FUNDING INFORMATION: The study was designed under the responsibility of Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA, in conjunction with the steering committee. The study was funded by Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA. Pembrolizumab was provided by Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA. All authors had full access to all of the data in the study and had final responsibility for the decision to submit for publication. ACKNOWLEDGMENTS: This work was funded by Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA. We thank the patients and their families and caregivers for participating in this trial, and all investigators and site personnel. Medical writing and editorial assistance were provided by Jemimah Walker, PhD, Mehak Aggarwal, PharmD, and Doyel Mitra, PhD, CMPP, of ApotheCom (Yardley, PA, USA). This assistance was funded by Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA

Análisis de diferentes ensilados del alga Saccharina latissima

1 página.- Trabajo presentado a las: XIX Jornadas sobre Producción Animal AIDA. Zaragoza, España, 1-2 junio 2021. Congreso Virtual.Trabajo realizado en el marco del Proyecto ES 631289 “Ensiled Cultivated Macroalgae as a Sustainable Ruminant Feedstuff (EnMac)”, financiado por el programa MABIT (Marine Biotechnology in Northern Norway) de Norueg

Effectiveness and safety of aflibercept for metastatic colorectal cancer: retrospective review within an early access program in Spain

[Purpose] In the VELOUR study, aflibercept + FOLFIRI regimen resulted in improved survival in metastatic colorectal cancer (mCRC) patients who progressed after oxaliplatin. The use of aflibercept outside the clinical trial framework needs to be further assessed in terms of effectiveness and tolerability.[Methods] Early access to aflibercept through a named patient programme (NPP) was provided to mCRC patients receiving FOLFIRI as second-line treatment in Spain. The effectiveness of aflibercept was assessed as progression-free survival (PFS) achieved within the NPP population. Post hoc analyses on PFS were done according to certain baseline characteristics (K-RAS mutation, prior targeted therapy) or prognostic factors.[Results] Registries from 71 mCRC patients included in the NPP were reviewed retrospectively. The median age for the NPP population was 64 years (19.7 % aged ≥70 years) and 63.4 % patients had ≥2 metastases. A median PFS of 5.3 months (95 % CI, 3.6–8.5 months) was achieved, which did not depend on K-RAS mutation status or prior targeted therapy received. The risk of progression or death increased in patients with a poor prognosis as per the GERCOR score (performance status [PS] 1–2 and increased baseline lactate dehydrogenase [LDH] level) compared with patients with a good prognosis (PS 0 and normal LDH level) (median PFS: 2.6 vs. 8.3 months, respectively; p = 0.0124). Aflibercept was well tolerated, with a manageable toxicity profile.[Conclusions] Bearing in mind the differences in sample size, the PFS achieved with the aflibercept + FOLFIRI regimen in the real-life practice setting is comparable to that observed in the clinical trial setting.This study was supported by Sanofi. A. Del Campo (Pivotal SL) provided medical writing services on behalf of Sanofi Pharmaceuticals

Feeding mango wastes to dairy goats: Effects on diet digestibility, ruminal fermentation, and milk yield and composition

The aim of this study was to assess the potential of mango wastes (0.65:0.35 pulp:peels mixture, PP) preserved in multinutrient blocks (MMB) for goats feeding that replaced part of a commercial concentrate for dairy goats, and contained 0.29 of the mango PP. The multinutrient blocks were formulated to have similar protein content than the concentrate. In addition, soybean meal was totally replaced with sunflower meal. Two diets composed of 0.4 of alfalfa hay and either 0.6 of a commercial concentrate (control diet) or 0.4 of concentrate and 0.2 of MMB (MAN diet) were fed to two homogeneous groups of Murciano-Granadina dairy goats of 8 animals each. Feed intake, diet digestibility, milk yield, composition and fatty acid (FA) profile, ruminal fermentation parameters, and urinary excretion of purine derivatives were measured. The daily intake of MMB averaged 83.9 g dry matter (DM) per goat and was accompanied by an average daily decrease of 71.0 g in concentrate intake (P = 0.324) and 41.0 g of increase in alfalfa hay intake. However, there were no differences between groups (P ≥ 0.253) in the intake of total DM, nitrogen and gross energy. Similarly, nutrient digestibility, utilization of both nitrogen (N) and energy, and urinary excretion of purine derivatives were similar (P ≥ 0.103) for both diets. Partial replacement of concentrate with MMB did not affect (P ≥ 0.358) milk yield and composition, but the milk from MAN-fed goats had greater (P = 0.038) saturated FA and lower (P = 0.047) polyunsaturated FA proportions than the milk from goats fed the control diet. Feeding the MAN diet also resulted in greater (P = 0.003) proportions of milk FA with less than 16 carbons and lower (P = 0.009) proportions of FA with more than 16 carbons compared with feeding the control diet. Similar values (P ≥ 0.213) of ruminal pH and concentrations of NH-N and total volatile fatty acids were registered in both groups of animals, but goats fed the MAN diet had lower (P = 0.003) acetate and greater (P = 0.029) propionate proportions than those fed the control diet. Under the circumstances of the present study, the profit of milk production (€/d) tended (P = 0.056) to be greater for the MAN diet compared with the control diet. Although multinutrient blocks seem to be a feasible option to preserve mango wastes, more studies are needed to increase the intake of MMB by goats.Funding was provided by the Excellence Programme of Junta de Andalucía (Project P12-AGR-587)