Resultados preliminares del estudio piloto de radioterapia Intraoperatoria para el cáncer de mama con fotones de 70KeV"

Introducción: La radioterapia intraoperatoria (RIO) es una nueva modalidad que permite la administración de una única dosis de radiación ionizante sobre el lecho tumoral durante la intervención quirúrgica. En el tratamiento adyuvante del cáncer de mama puede aplicarse como monoterapia o como refuerzo de la radioterapia externa. Diversos estudios han evaluado si la RIO con 50KeV es una técnica factible como monoterapia adyuvante en pacientes con estadios precoces de cáncer de mama. Por lo pronto se ha demostrado la seguridad de la técnica y se ha demostrado su no inferioridad respecto a otras. Objetivo: El principal objetivo del estudio es describir y evaluar un protocolo para la aplicación segura de RIO a 70 KeV como monoterapia adyuvante. También se describirán los efectos agudos y subagudos. Diseño del estudio: Consiste en un estudio observacional retrospectivo unicéntrico. Los criterios de inclusión han sido extraídos del protocolo establecido de radioterapia intraoperatoria en el cáncer de mama: mujeres de >60 años con cáncer de mama en estadio precoz y sin factores pronósticos desfavorables. Los datos obtenidos durante el seguimiento de las pacientes han sido recogidos en el SAP. Material y métodos: La unidad del tratamiento consiste en un acelerador miniaturizado de electrones con una tensión máxima de 70kV. Cuenta con cinco aplicadores de diferente tamaño (3’5-5’5 cm) que se eligen en función del tamaño de la cavidad tras la tumorectomía. Para establecer la dosimetría se realizaron diversas simulaciones en medio acuoso. En el seguimiento de las pacientes se ha valorado la presencia de dermitis en las revisiones rutinarias, se ha realizado un TAC de tórax a las 7 semanas y una mamografía de control a las 13 semanas de la intervención. Resultados: La media de seguimiento fueron 9.6 meses. De las 17 pacientes que fueron incluidas, cuatro presentaron ganglios positivos, por lo que requirieron radioterapia externa posterior. En otras cuatro pacientes no se pudo aplicar RIO tras la intervención por no terminar de cumplir los criterios necesarios. Ninguna ha presentado recurrencia ni neumonitis durante el seguimiento. Cinco han presentado eritema leve grado 1 y una grado 2. Discusión: No se han observado incidencias operativas. El número de pacientes que han necesitado radioterapia posterior ha sido esperable y además asumible. Por cuestiones anatómicas, cuatro pacientes no fueron subsidiarias de RIO. Se ha establecido que la dosis de 12’5 Gy a 0’5 cm es segura. Conclusiones: El protocolo descrito es capaz de administrar RIO para cáncer de mama de manera reproductible, segura y relativamente sencilla. Los únicos efectos secundarios agudos observados han sido dermatitis leves, sin transcendencia en el performance status de las pacientes. No se han observado recidivas a corto plazo. Es necesario un periodo de tiempo más prolongado para valorar la efectividad de la técnicaIntroduction: Intraoperative Radiation Therapy (IORT) is the latest technique on the treatment of breast cancer, that consists in emitting a high dose of radiation in the tumour bed at the same time of the intervention. It is useful to treat breast cancer as monotherapy or as boost before external radiotherapy. This technique, using 50KeV, has already been evaluated in other studies and its conclusion is that it is effective and safe in the management of early stages of breast cancer. Background: The main objective is to describe and evaluate a protocol to set up a safe dose of IORT 70 KeV as adyuvant monotherapy. It is also important to recognize and describe its acute and subacute effects. Study design: It is a single-center descriptive restrospective observational study. The inclusion criteria are: women aged 60 with early stages of breast cancer and with good prognosis. The information was obtained in the data base of the CHUC known as SAP. Methods: The treatment consists of a miniature linear particle accelerator that emits an average energy of electron at 70 KeV. It has five applicators that are choosen according to the size of the remaining cavity after the surgery. Simulations were done in aqueous medium to determinate the doses. During follow-up we evaluated the skin in order to find radiodermatitis. A scanner and a mammography were also done the seventh and thirteenth week after surgery, respectively. Results: The median time of follow-up was 9.6 months. 17 patients were included in the study. Four of them had a positive sentinel node and required external radiotherapy. Four patients did not meet the inclusion criteria for IORT after the surgery. None of them developed local recurrence or pneumonitis. According to RTOG Scoring Criteria, five had erythema stage 1 and only one had stage 2. Discussion: No incidents related to the techniche were observed. The quantity of patients that required external radiotherapy was expected and manageable. Four patients could not receive IORT because the remaining cavity after the surgery did not meet the inclusión criteria of the study. Dosis of 12’5 Gy at 0’5 cm has been established as safe. Conclusions: The protocol described to administrate IORT is safe, simple and can easily be replicated. The only acute effects that were observed were mild dermatitis without changes in the performance status of the patients. There was not local recurrence. Longer follow-up studies are needed to stablish the effectiveness of the techniche

Mitochondrial glutathione: Features, regulation and role in disease

El pdf del artículo es la versión post-print.[Background]: Mitochondria are the powerhouse of mammalian cells and the main source of reactive oxygen species (ROS) associated with oxygen consumption. In addition, they also play a strategic role in controlling the fate of cells through regulation of death pathways. Mitochondrial ROS production fulfills a signaling role through regulation of redox pathways, but also contributes to mitochondrial damage in a number of pathological states. [Scope of review]: Mitochondria are exposed to the constant generation of oxidant species, and yet the organelle remains functional due to the existence of an armamentarium of antioxidant defense systems aimed to repair oxidative damage, of which mitochondrial glutathione (mGSH) is of particular relevance. Thus, the aim of the review is to cover the regulation of mGSH and its role in disease. [Major conclusions]: Cumulating evidence over recent years has demonstrated the essential role for mGSH in mitochondrial physiology and disease. Despite its high concentration in the mitochondrial matrix, mitochondria lack the enzymes to synthesize GSH de novo, so that mGSH originates from cytosolic GSH via transport through specific mitochondrial carriers, which exhibit sensitivity to membrane dynamics. Depletion of mGSH sensitizes cells to stimuli leading to oxidative stress such as TNF, hypoxia or amyloid β-peptide, thereby contributing to disease pathogenesis. [General significance]: Understanding the regulation of mGSH may provide novel insights to disease pathogenesis and toxicity and the opportunity to design therapeutic targets of intervention in cell death susceptibility and disease. This article is part of a Special Issue entitled Cellular functions of glutathione. © 2012 Elsevier B.V.The work was supported by grants: SAF2009-11417, SAF2010-15760, and SAF2011-23031 (Plan Nacional de I + D), Proyectos de Investigación en Salud PI10/02114 and PS09/00056 (Instituto de Salud Carlos III), P50-AA-11999 (Research Center for Liver and Pancreatic Diseases, US National Institute on Alcohol Abuse and Alcoholism) and by CIBEREHD from the Instituto de Salud Carlos III.Peer Reviewe

Leishmania infantum-specific IFN-γ production in stimulated blood from dogs with clinical leishmaniosis at diagnosis and during treatment

There is limited data regarding Leishmania infantum specific T cell mediated immunity in naturally infected sick dogs at the time of diagnosis and during anti-Leishmania treatment. Our aim was to investigate the kinetics of L. infantum specific IFN-γ production in dogs with leishmaniosis at the time of diagnosis and during treatment and to correlate it with specific L. infantum antibodies, blood parasitemia and clinicopathological findings. Thirty-four dogs were diagnosed with leishmaniosis based on physical examination, routine laboratory tests and L. infantum-specific antibody levels by quantitative ELISA. Heparinized whole blood was stimulated with L. infantum soluble antigen (LSA) and concanavalin A (ConA) and incubated for 5 days. IFN-γ concentration was evaluated in supernatants of stimulated blood using a commercial sandwich ELISA. Leishmania real-time PCR was also performed for assessing blood parasitemia. Dogs were treated with meglumine antimoniate and allopurinol. Sixteen dogs were classified as IFN-γ non-producers after LSA stimulation (mean ± SD: 0 ± 0 pg/mL) and 18 dogs as IFN-γ producers (mean ± SD: 2885.3 ± 4436.1 pg/mL) at the time of diagnosis (P < 0.0001). IFN-γ non-producers were classified in a more severe clinical staging than IFN-γ producers that presented a mild to moderate clinical staging (P = 0.03). In the IFN-γ non-producer group, production of IFN-γ after LSA stimulation was significantly increased during treatment especially at day 365 (P = 0.018) together with clinical improvement when compared with day 0. In contrast, IFN-γ producers maintained their IFN-γ production after LSA stimulation and no statistically significant changes were found during treatment follow-up. At diagnosis, IFN-γ non-producers showed a significantly higher blood parasitemia versus IFN-γ -producers (P = 0.005). IFN-γ non-producers drastically reduced blood parasitemia to minimum values at day 365 when compared with day 0 (P = 0.017). No significant differences were found at day 365 in blood parasitemia of IFN-γ producers compared to pre-treatment. At diagnosis, L. infantum specific antibodies were higher in IFN-γ non-producers than IFN-γ producers (P = 0.014). A marked reduction of antibody levels was found at day 365 when compared with day 0 in IFN-γ non-producers (P = 0.005) and producers (P = 0.001). These results demonstrate that IFN-γ concentration increases with long-term anti-Leishmania treatment together with clinical improvement in dogs that do not produce IFN-γ at diagnosis. Together with clinical recovery, reduction in blood parasitemia and L. infantum specific antibodies, tracking IFN-γ concentration could constitute an important prognostic tool for immune monitoring in CanL

Human exposure and risk assessment of PAHs bound to three PM fractions (10, 2.5 and 1) in an area influenced by a cement plant

In the present study, we evaluated the concentrations of PAHs in 3 PM fractions (10, 2.5 and 1) collected in the surroundings of a cement plant located in Barcelona. PAH content and speciation were developed for the three fractions to elucidate their distribution among different sizes. Complementarily, the human health risks associated to the PAH exposure were risks by considering the daily activity pattern of an average adult living in of Barcelona (Spain).Financial support was received by the Spanish Ministry of Economy and Competitiveness (MINECO), through the project CTM2012-32778. F. Sánchez-Soberón received a doctoral scholarship as part of the project above mentioned. We also want to thank the European Union Seventh Framework Program for the funding received through the HEALS project (grant agreement No. 603946).Peer reviewe

Cysteine cathepsins control hepatic NF-κB-dependent inflammation via sirtuin-1 regulation

Sirtuin-1 (SIRT1) regulates hepatic metabolism but its contribution to NF-κB-dependent inflammation has been overlooked. Cysteine cathepsins (Cathepsin B or S, CTSB/S) execute specific functions in physiological processes, such as protein degradation, having SIRT1 as a substrate. We investigated the roles of CTSB/S and SIRT1 in the regulation of hepatic inflammation using primary parenchymal and non-parenchymal hepatic cell types and cell lines. In all cells analyzed, CTSB/S inhibition reduces nuclear p65-NF-κB and κB-dependent gene expression after LPS or TNF through enhanced SIRT1 expression. Accordingly, SIRT1 silencing was sufficient to enhance inflammatory gene expression. Importantly, in a dietary mouse model of non-alcoholic steatohepatitis, or in healthy and fibrotic mice after LPS challenge, cathepsins as well as NF-κB-dependent gene expression are activated. Consistent with the prominent role of cathepsin/SIRT1, cysteine cathepsin inhibition limits NF-κB-dependent hepatic inflammation through the regulation of SIRT1 in all in vivo settings, providing a novel anti-inflammatory therapeutic target in liver disease.This study was supported by grants from the Instituto de Salud Carlos III (PI13/00374 to MM), Ministerio de Economía y Competitividad (SAF2015-69944-R to JFC, SAF2013-47246-R to AC, SAF2015-66515-R to AM) and co-funded by FEDER (Fondo Europeo de Desarrollo Regional, Unión Europea. “Una manera de hacer Europa”); center grant P50-AA-11999 from Research Center for Liver and Pancreatic Diseases (US-NIAAA to JFC); and by CIBERehd. AT is a recipient of a FPU fellowship recipient from the Ministerio de Educación, Cultura y Deporte.Peer Reviewe

Angiogenin secretion from hepatoma cells activates hepatic stellate cells to amplify a self-sustained cycle promoting liver cancer

Hepatocellular carcinoma (HCC) frequently develops in a pro-inflammatory and pro-fibrogenic environment with hepatic stellate cells (HSCs) remodeling the extracellular matrix composition. Molecules secreted by liver tumors contributing to HSC activation and peritumoral stromal transformation remain to be fully identified. Here we show that conditioned medium from HCC cell lines, Hep3B and HepG2, induced primary mouse HSCs transdifferentiation, characterized by profibrotic properties and collagen modification, with similar results seen in the human HSC cell line LX2. Moreover, tumor growth was enhanced by coinjection of HepG2/LX2 cells in a xenograft murine model, supporting a HCC-HSC crosstalk in liver tumor progression. Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules. In fact, recombinant angiogenin induced in vitro HSC activation requiring its nuclear translocation and rRNA transcriptional stimulation. Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin. Finally, neomycin administration reduced tumor growth of HepG2-LX2 cells coinjected in mice. In conclusion, angiogenin secretion by HCCs favors tumor development by inducing HSC activation and ECM remodeling. These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC managementThis study was funded by grants from the Instituto de Salud Carlos III (FIS PI12/00110, PI09/00056 to A.M., FIS PI10/02114, PI13/00374 to M.M., PI12/01265 to J.C. and PI11/0325 to J.F.C.), Ministerio de Economía y Competitividad (SAF 2012/34831 to J.F.C. and SAF2011-23031 to C.G.R.) and co-funded by FEDER (Fondo Europeo de Desarrollo Regional, Unión Europea. “Una manera de hacer Europa”); center grant P50-AA-11999 from Research Center for Liver and Pancreatic Diseases, US NIAAA to J.F.C.); Fundació la Marató de TV3 to J.F.C., Mutua Madrilea (AP103502012) to C.G.R., and by CIBERehd from the Instituto de Salud Carlos IIIPeer Reviewe

Ghrelin attenuates hepatocellular injury and liver fibrogenesis in rodents and influences fibrosis progression in humans

El pdf del artículo es la versión pre-print.-- et al-There are no effective antifibrotic therapies for patients with liver diseases. We performed an experimental and translational study to investigate whether ghrelin, an orexigenic hormone with pleiotropic properties, modulates liver fibrogenesis. Recombinant ghrelin was administered to rats with chronic (bile duct ligation) and acute (carbon tetrachloride) liver injury. Hepatic gene expression was analyzed by way of microarray analysis and quantitative polymerase chain reaction. The hepatic response to chronic injury was also evaluated in wild-type and ghrelin-deficient mice. Primary human hepatic stellate cells were used to study the effects of ghrelin in vitro. Ghrelin hepatic gene expression and serum levels were assessed in patients with chronic liver diseases. Ghrelin gene polymorphisms were analyzed in patients with chronic hepatitis C. Recombinant ghrelin treatment reduced the fibrogenic response, decreased liver injury and myofibroblast accumulation, and attenuated the altered gene expression profile in bile duct-ligated rats. Moreover, ghrelin reduced the fibrogenic properties of hepatic stellate cells. Ghrelin also protected rats from acute liver injury and reduced the extent of oxidative stress and inflammation. Ghrelin-deficient mice developed exacerbated hepatic fibrosis and liver damage after chronic injury. In patients with chronic liver diseases, ghrelin serum levels decreased in those with advanced fibrosis, and ghrelin gene hepatic expression correlated with expression of fibrogenic genes. In patients with chronic hepatitis C, polymorphisms of the ghrelin gene (994CT and 604GA) influenced the progression of liver fibrosis. Conclusion: Ghrelin exerts antifibrotic effects in the liver and may represent a novel antifibrotic therapy. Copyright © 2010 by the American Association for the Study of Liver Diseases.Supported by grants from the Ministerio de Ciencia e Investigación (SAF2005-06245), from the Instituto de Salud Carlos III (FIS2005-050567, FIS 2008-PI08/0237 and PI070497), and from the European Community FP6 (LSHB-CT-2007-036644 - DIALOK) and by fellowships from Institut d’Investigacions Biomèdiques August Pi i Sunyer (to M. M. and M. D.), the Fundación Bilbao Vizcaya Argentaria (to M. D.) and the Fundació Clínic (to P. S. B.).Peer Reviewe

Regorafenib Alteration of the BCL-xL/MCL-1 Ratio Provides a Therapeutic Opportunity for BH3-Mimetics in Hepatocellular Carcinoma Models

Background: The multikinase inhibitor regorafenib, approved as second-line treatment for hepatocellular carcinoma (HCC) after sorafenib failure, may induce mitochondrial damage. BH3-mimetics, inhibitors of specific BCL-2 proteins, are valuable drugs in cancer therapy to amplify mitochondrial-dependent cell death. Methods: In in vitro and in vivo HCC models, we tested regorafenib's effect on the BCL-2 network and the efficacy of BH3-mimetics on HCC treatment. Results: In hepatoma cell lines and Hep3B liver spheroids, regorafenib cytotoxicity was potentiated by BCL-xL siRNA transfection or pharmacological inhibition (A-1331852), while BCL-2 antagonism had no effect. Mitochondrial outer membrane permeabilization, cytochrome c release, and caspase-3 activation mediated A-1331852/regorafenib-induced cell death. In a patient-derived xenograft (PDX) HCC model, BCL-xL inhibition stimulated regorafenib activity, drastically decreasing tumor growth. Moreover, regorafenib-resistant HepG2 cells displayed increased BCL-xL and reduced MCL-1 expression, while A-1331852 reinstated regorafenib efficacy in vitro and in a xenograft mouse model. Interestingly, BCL-xL levels, associated with poor prognosis in liver and colorectal cancer, and the BCL-xL/MCL-1 ratio were detected as being increased in HCC patients. Conclusion: Regorafenib primes tumor cells to BH3-mimetic-induced cell death, allowing BCL-xL inhibition with A-1331852 or other strategies based on BCL-xL degradation to enhance regorafenib efficacy, offering a novel approach for HCC treatment, particularly for tumors with an elevated BCL-xL/MCL-1 ratio

Gas6, una proteína dependiente de la vitamina k implicada en la respuesta a la infección por sars-cov-2

Trabajo presentado en el LXIII Congreso Nacional SEHH/ XXXVII Congreso Nacional SETH, celebrado en Pamplona (Navarra) del 14 al 16 de octubre de 2021

A Nutraceutical Rich in Docosahexaenoic Acid Improves Portal Hypertension in a Preclinical Model of Advanced Chronic Liver Disease

Inflammation and oxidative stress play a key role in the pathophysiology of advanced chronic liver disease (ACLD) and portal hypertension (PH). Considering the current lack of effective treatments, we evaluated an anti-inflammatory and antioxidant nutraceutical rich in docosahexaenoic acid (DHA) as a possible therapy for ACLD. We investigated the effects of two-week DHA supplementation (500 mg/kg) on hepatic fatty acids, PH, oxidative stress, inflammation, and hepatic stellate cell (HSC) phenotype in rats with ACLD. Additionally, the effects of DHA were evaluated in murine macrophages and human HSC. In contrast to vehicle-treated animals, cirrhotic rats receiving DHA reestablished a healthy hepatic fatty acid profile, which was associated with an improvement in PH. The mechanisms underlying this hemodynamic improvement included a reduction in oxidative stress and inflammation, as well as a marked HSC deactivation, confirmed in human HSC. Experiments with cultured macrophages showed that treatment with DHA protects against pro-inflammatory insults. The present preclinical study demonstrates that a nutraceutical rich in DHA significantly improves PH in chronic liver disease mainly by suppressing inflammation and oxidative stress-driven HSC activation, encouraging its evaluation as a new treatment for PH and cirrhosis