Dietary (poly)phenols are extensively metabolized, limiting their anticancer activity. Exosomes (EXOs) are extracellular vesicles that could protect polyphenols from metabolism. Our objective was to ...compare the delivery to breast tissue and anticancer activity in breast cancer cell lines of free curcumin (CUR) and resveratrol (RSV) vs. their encapsulation in milk-derived EXOs (EXO-CUR and EXO-RSV). A kinetic breast tissue disposition was performed in rats. CUR and RSV were analyzed using UPLC-QTOF-MS and GC-MS, respectively. Antiproliferative activity was tested in MCF-7 and MDA-MB-231 breast cancer and MCF-10A non-tumorigenic cells. Cell cycle distribution, apoptosis, caspases activation, and endocytosis pathways were determined. CUR and RSV peaked in the mammary tissue (41 ± 15 and 300 ± 80 nM, respectively) 6 min after intravenous administration of EXO-CUR and EXO-RSV, but not with equivalent free polyphenol concentrations. Nanomolar EXO-CUR or EXO-RSV concentrations, but not free CUR or RSV, exerted a potent antiproliferative effect on cancer cells with no effect on normal cells. Significant (p < 0.05) cell cycle alteration and pro-apoptotic activity (via the mitochondrial pathway) were observed. EXO-CUR and EXO-RSV entered the cells primarily via clathrin-mediated endocytosis, avoiding ATP-binding cassette transporters (ABC). Milk EXOs protected CUR and RSV from metabolism and delivered both polyphenols to the mammary tissue at concentrations compatible with the fast and potent anticancer effects exerted in model cells. Milk EXOs enhanced the bioavailability and anticancer activity of CUR and RSV by acting as Trojan horses that escape from cancer cells’ ABC-mediated chemoresistance.
Objetivo: Presentar una nueva técnica quirúrgica y los resultados clínicos comparativos de pacientes con roturas de tenorrafia primaria o lesiones crónicas del flexor profundo en las zonas I y II de ...Verdan, tratados con avance tendinoso no convencional. Materiales y Métodos: Se incluyó a pacientes >18 años, con antecedente de lesiones en las zonas I y II de Verdan y un seguimiento mínimo de 36 meses. La serie estaba formada por 13 pacientes (edad promedio 29 años) que fueron divididos en dos grupos según el tipo de lesión (7 con roturas de tenorrafia primaria y 6 con lesiones crónicas del flexor profundo) y la técnica quirúrgica utilizada (alargamiento en Z más sutura término-terminal y sutura lateral del tendón alargado a un flexor vecino indemne en la zona V, respectivamente). Se empleó la Clasificación de Strickland para la evaluación. Resultados: El seguimiento promedio fue de 51 meses. El intervalo promedio entre la lesión y la cirugía fue de 11.7 semanas. Según la Clasificación de Strickland, 8 pacientes tuvieron resultados excelentes; 4, buenos y uno, pobre. El avance tendinoso promedio fue de 20,5 mm en ambos grupos. Conclusiones: El avance tendinoso no convencional para lesiones en las zonas I y II de Verdan, sea en roturas de tenorrafia primaria o lesiones crónicas del flexor profundo, resultó un tratamiento reproducible y eficaz. Palabras clave: Tendón flexor; alargamiento; zetaplastia.Nivel de Evidencia: III
Donación en asistolia controlada: cómo iniciar un programa Villar-García, Susana; Martín-López, Carlos E.; Pérez-Redondo, Marina ...
Cirugía cardiovascular,
November-December 2022, 2022-11-00, 2022-11-01, Volume:
29, Issue:
6
Journal Article
Peer reviewed
Open access
La donación en asistolia controlada se ha convertido en una de las principales estrategias para aumentar el número de donantes. Hasta el momento el corazón de este tipo de donantes no era valorado ...por el riesgo de lesión isquémica durante la limitación del soporte vital. En los últimos años, con los sistemas de perfusión ex situ se iniciaron programas de donación cardiaca en asistolia controlada que demostraron la viabilidad miocárdica. El uso de perfusión regional normotérmica en nuestro medio y la experiencia de otros países en la recuperación cardiaca nos lleva a elaborar un protocolo de extracción cardiaca en asistolia controlada con valoración in situ y conservación en frío.
Se conforma un equipo multidisciplinar que realiza un protocolo de extracción cardiaca con donante en asistolia controlada. Tras una fase de experimentación animal, dicho protocolo es aprobado por el Comité de Ética para la Asistencia Sanitaria y por la Organización Nacional de Trasplantes.
Se realiza un protocolo que establece los criterios de selección de donantes y receptores y que regula el procedimiento de donación con perfusión toracoabdominal tras exclusión de los troncos supraaórticos. La valoración cardiaca es in situ y la conservación estática, en frío. Este protocolo se comunica al resto de hospitales donantes, y esto nos lleva a realizar el primer trasplante cardiaco en España con asistolia controlada.
El trasplante cardiaco con donación en asistolia controlada es una realidad clínica que permitirá aumentar el número de potenciales donantes. Las características técnicas del procedimiento hacen que este tipo de donación se deba concentrar en centros con experiencia.
Donation after circulatory death has become one of the main strategies to increase the number of donors. Until now, the heart of this type of donor was not evaluated due to the risk of ischaemic injury during the withdrawal of life support. In recent years, ex situ perfusion systems have initiated cardiac donation programmes after circulatory death that have demonstrated myocardial viability. The use of normothermic regional perfusion in our environment and the experience of other countries in cardiac recovery led us to develop a protocol for cardiac procurement in donation after circulatory death with in situ assessment and cold preservation.
A multidisciplinary team was formed to create a cardiac extraction protocol with donation after circulatory death. After a phase of animal experimentation, the protocol was approved by the Ethics Committee for Health Care and by the National Transplant Organization.
A protocol was drawn up establishing the criteria for donor and recipient selection and regulating the donation procedure with thoracic-abdominal perfusion after exclusion of the supra-aortic trunks. Cardiac evaluation is in situ and static preservation in cold storage. This protocol was sent to the rest of the donor hospitals and this led us to perform the first heart transplant in Spain with this donation modality.
Cardiac transplantation with donation after circulatory death is a clinical reality that will increase the number of potential donors. The technical characteristics of the procedure mean that this type of donation should be performed in experienced centres.
Desde el 8 de febrero de 2021, la Sociedad Española de Cirugía Cardiovascular y Endovascular (SECCE) puso en marcha el Registro Español de Cirugía Cardiaca (RECC) que está disponible para las ...diferentes unidades de cirujanos cardiovasculares de nuestro país. Es una herramienta que permite recopilar datos de pacientes sometidos a cirugía cardiaca, vascular o endovascular. Tras dos años de desarrollo, hemos llevado a cabo un análisis de la calidad de la información obtenida para adquirir una visión general de su contenido.
La información ha sido analizada de forma anónima a nivel de paciente, hospital y provincia. Para la estimación de la mortalidad ajustada por riesgo se utilizó la escala de estimación de riesgo preoperatorio EuroSCORE II.
Se han incluido en el RECC un total de 7.087 intervenciones, de las cuales 6.267 se trataban de cirugías cardiacas mayores. Del total de intervenciones mayores, 53,9% eran cirugías valvulares, 25,2% de revascularización miocárdica y 14,9% de aorta. La mortalidad global de la serie fue de 5,0% y el índice de mortalidad ajustada al riesgo (IMAR) de 0,88. La calibración del EuroSCORE II en la muestra global fue buena en los pacientes de riesgo más bajo, aunque sobreestimó la mortalidad en los de alto riesgo.
El RECC se trata de una base de datos clínica nacional que permite el análisis de datos de pacientes con el fin de evaluar de forma precisa el volumen de la actividad, riesgo y resultados. A nivel local, podría utilizarse como una herramienta para mejorar la calidad de la atención y el desarrollo de programas correctivos.
Since February 8, 2021, the Spanish Society of Cardiovascular and Endovascular Surgery got under way the Spanish Registry of Cardiac Surgery (RECC), which is available for the different units of cardiovascular surgeons in our country. It is a tool that allows collect patient-level data of patients undergoing cardiac, vascular or endovascular surgery. After two years of development, we have carried out an analysis of the quality of the information obtained in order to acquire an overview of its content.
The information has been analyzed anonymously at patient, hospital and province level. For risk-adjusted mortality estimation, the EuroSCORE II preoperative risk estimation scale was used.
A total of 7087 interventions have been included. Six thousand two hundred and sixty-seven were major cardiac surgeries: 53.9% valvular, 25.2% coronary artery bypass grafting, and 14.9% aortic procedures. The overall mortality was 5.0% and the risk-adjusted mortality rate was 0.88. The EuroSCORE II calibration in the overall sample was good in the lowest-risk patients, although it overestimated mortality in high-risk patients.
RECC is a nationally defined clinical database in the field of cardiovascular surgery. RECC allows a patient-level data analysis in order to perform an accurate analysis of the volumen of activity, risk adjustment and results. Locally, it could be used as a tool to improve the quality of care and development of corrective programs.
Marfan syndrome (MFS) is an autosomal dominant disorder of the connective tissue caused by mutations in the
(fibrillin-1) gene encoding a large glycoprotein in the extracellular matrix called ...fibrillin-1. The major complication of this connective disorder is the risk to develop thoracic aortic aneurysm. To date, no effective pharmacologic therapies have been identified for the management of thoracic aortic disease and the only options capable of preventing aneurysm rupture are endovascular repair or open surgery. Here, we have studied the role of mitochondrial dysfunction in the progression of thoracic aortic aneurysm and mitochondrial boosting strategies as a potential treatment to managing aortic aneurysms.
Combining transcriptomics and metabolic analysis of aortas from an MFS mouse model (
) and MFS patients, we have identified mitochondrial dysfunction alongside with mtDNA depletion as a new hallmark of aortic aneurysm disease in MFS. To demonstrate the importance of mitochondrial decline in the development of aneurysms, we generated a conditional mouse model with mitochondrial dysfunction specifically in vascular smooth muscle cells (VSMC) by conditional depleting Tfam (mitochondrial transcription factor A;
mice). We used a mouse model of MFS to test for drugs that can revert aortic disease by enhancing Tfam levels and mitochondrial respiration.
The main canonical pathways highlighted in the transcriptomic analysis in aortas from
mice were those related to metabolic function, such as mitochondrial dysfunction. Mitochondrial complexes, whose transcription depends on Tfam and mitochondrial DNA content, were reduced in aortas from young
mice. In vitro experiments in
-silenced VSMCs presented increased lactate production and decreased oxygen consumption. Similar results were found in MFS patients. VSMCs seeded in matrices produced by Fbn1-deficient VSMCs undergo mitochondrial dysfunction. Conditional Tfam-deficient VSMC mice lose their contractile capacity, showed aortic aneurysms, and died prematurely. Restoring mitochondrial metabolism with the NAD precursor nicotinamide riboside rapidly reverses aortic aneurysm in
mice.
Mitochondrial function of VSMCs is controlled by the extracellular matrix and drives the development of aortic aneurysm in Marfan syndrome. Targeting vascular metabolism is a new available therapeutic strategy for managing aortic aneurysms associated with genetic disorders.
Increased collagen cross-linking (CCL) has been described in hypertensive cardiomyopathy by means of reduced serum ratio of serum carboxyterminal telopeptide of collagen type I (CITP) to matrix ...metalloproteinase-1 (MMP1). Previous studies have demonstrated the existence of primary impaired diastole in patients with Marfan syndrome (MFS), but little is known about the pathophysiology of this condition.
60 MFS patients (without previous cardiovascular surgery or significant valvular regurgitation) and 24 healthy controls (age and sex-matched) were enrolled. All participants underwent a comprehensive transthoracic echocardiographic study, including left atrial and left ventricular speckle-tracking strain analysis. CITP and MMP1 were measured in peripheral blood.
All participants had normal diastolic function according to guidelines. Peak left atrial strain in the reservoir phase (LASr) was significantly reduced in the MFS cohort compared to controls (32.2 ± 9.4 vs 43.9 ± 7.0%; p < 0.001). Serum CITP and CITP:MMP1 ratio were lower among MFS patients, showing significant correlations with LASr (R = 0.311; p = 0.020 and R = 0.437; p = 0.001, respectively). The MFS cohort was divided into quartiles of LASr. MFS patients in the lowest quartile of LASr (<26%) had significantly lower values of CITP:MMP1 ratio compared to the other quartiles.
The analysis of serum biomarkers revealed the presence of increased CCL in association with reduced LASr in the MFS cohort. Our results suggest that excessive CCL may play a role in the development of primary myocardial impairment in these patients. Future studies are needed to confirm our findings and evaluate the prognostic role of CCL markers in the MFS population.
•Left atrial strain is reduced in patients with Marfan syndrome (MFS), reflecting a subtle impairment in diastolic function•Lower CITP:MMP1 ratio is observed in the MFS cohort, reflecting a higher degree of collagen cross-linking (CCL).•Left atrial strain is significantly correlated with CCL biomarkers, with lower values of CITP:MMP1 in the lowest quartile of LASr.•These findings suggest that CCL may play a role in the pathophysiology of MFS-related cardiomyopathy.
Thoracic aortic aneurysm, as occurs in Marfan syndrome, is generally asymptomatic until dissection or rupture, requiring surgical intervention as the only available treatment. Here, we show that ...nitric oxide (NO) signaling dysregulates actin cytoskeleton dynamics in Marfan Syndrome smooth muscle cells and that NO-donors induce Marfan-like aortopathy in wild-type mice, indicating that a marked increase in NO suffices to induce aortopathy. Levels of nitrated proteins are higher in plasma from Marfan patients and mice and in aortic tissue from Marfan mice than in control samples, indicating elevated circulating and tissue NO. Soluble guanylate cyclase and cGMP-dependent protein kinase are both activated in Marfan patients and mice and in wild-type mice treated with NO-donors, as shown by increased plasma cGMP and pVASP-S239 staining in aortic tissue. Marfan aortopathy in mice is reverted by pharmacological inhibition of soluble guanylate cyclase and cGMP-dependent protein kinase and lentiviral-mediated Prkg1 silencing. These findings identify potential biomarkers for monitoring Marfan Syndrome in patients and urge evaluation of cGMP-dependent protein kinase and soluble guanylate cyclase as therapeutic targets.
Background
Systemic mastocytosis (SM) is a heterogeneous disease characterized by an expansion of KIT‐mutated mast cells (MC). KIT‐mutated MC display activated features and release MC mediators that ...might act on the tumour microenvironment and other immune cells. Here, we investigated the distribution of lymphocyte subsets in blood of patients with distinct subtypes of SM and determined its association with other disease features.
Methods
We studied the distribution of TCD4+ and TCD4− cytotoxic cells and their subsets, as well as total NK‐ and B cells, in blood of 115 SM patients—38 bone marrow mastocytosis (BMM), 67 indolent SM (ISM), 10 aggressive SM (ASM)‐ and 83 age‐matched healthy donors (HD), using spectral flow cytometry and the EuroFlow Immunomonitoring panel, and correlated it with multilineage KITD816V, the alpha‐tryptasemia genotype (HαT) and the clinical manifestations of the disease.
Results
SM patients showed decreased counts (vs. HD) of TCD4− cytotoxic cells, NK cells and several functional subsets of TCD4+ cells (total Th1, Th2‐effector memory, Th22‐terminal effector and Th1‐like Tregs), together with increased T‐follicular‐helper and Th1/Th17‐like Treg counts, associated with different immune profiles per diagnostic subtype of SM, in multilineal versus MC‐restricted KITD816V and in cases with a HαT+ versus HαT− genotype. Unique immune profiles were found among BMM and ISM patients with MC‐restricted KITD816V who displayed HαT, anaphylaxis, hymenoptera venom allergy, bone disease, pruritus, flushing and GI symptoms.
Conclusion
Our results reveal altered T‐ and NK‐cell immune profiles in blood of SM, which vary per disease subtype, the pattern of involvement of haematopoiesis by KITD816V, the HαT genotype and specific clinical manifestations of the disease.
Constitutively activated tumour MC in SM patients are associated with a broad activation of the innate immune response and an altered distribution of other adaptive immune cells. The altered circulating T‐ and NK‐cell immune profile in SM varies depending on disease subtype, the pattern of involvement of haematopoiesis by KITD816V, the HαT genotype and specific clinical manifestations of the disease. These altered immune profiles contribute to a better understanding of the different patterns of clinical manifestations of the disease, independently of the MC burden. Abbreviations: ASM, aggressive systemic mastocytosis; BMM, bone marrow mastocytosis; CM, central memory; EE, early effector; EM, effector memory; HD, healthy donor; HVA, Hymenoptera venom allergy; ISM, indolent systemic mastocytosis; KIT, receptor tyrosine kinase; MC, mast cell; NK, natural killer; SM, systemic mastocytosis; TE, terminal effector; TFH cell, T‐follicular cell; TM, transitional memory; Tregs, T‐regulatory cells.
Toca 511 (vocimagene amiretrorepvec), a nonlytic, amphotropic retroviral replicating vector (RRV), encodes and delivers a functionally optimized yeast cytosine deaminase (CD) gene to tumors. In ...orthotopic glioma models treated with Toca 511 and 5-fluorocytosine (5-FC) the CD enzyme within infected cells converts 5-FC to 5-fluorouracil (5-FU), resulting in tumor killing. Toca 511, delivered locally either by intratumoral injection or by injection into the resection bed, in combination with subsequent oral extended-release 5-FC (Toca FC), is under clinical investigation in patients with recurrent high-grade glioma (HGG). If feasible, intravenous administration of vectors is less invasive, can easily be repeated if desired, and may be applicable to other tumor types. Here, we present preclinical data that support the development of an intravenous administration protocol. First we show that intravenous administration of Toca 511 in a preclinical model did not lead to widespread or uncontrolled replication of the RVV. No, or low, viral DNA was found in the blood and most of the tissues examined 180 days after Toca 511 administration. We also show that RRV administered intravenously leads to efficient infection and spread of the vector carrying the green fluorescent protein (GFP)-encoding gene (Toca GFP) through tumors in both immune-competent and immune-compromised animal models. However, initial vector localization within the tumor appeared to depend on the mode of administration. Long-term survival was observed in immune-competent mice when Toca 511 was administered intravenously or intracranially in combination with 5-FC treatment, and this combination was well tolerated in the preclinical models. Enhanced survival could also be achieved in animals with preexisting immune response to vector, supporting the potential for repeated administration. On the basis of these and other supporting data, a clinical trial investigating intravenous administration of Toca 511 in patients with recurrent HGG is currently open and enrolling.
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