Cardiovascular tissue banking activity during SARS-CoV-2 pandemic: evolution of national protocols and Lombardy experience.

The worldwide pandemic outbreak due to severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) has created unprecedented challenges for public health services. Lombardy, region of the Northern Italy, has been the first area in the Western world whose organs and tissues procurement programs have had to face the virus pandemic emergency. We retrospectively collected and analyzed data about cardiovascular tissues (CT) in 2019 and in 2020. We aimed to describe the rapid evolution of SARS-CoV-2 regulation laws for tissue donor's selection and harvesting from February 2020 until January 2021. As expected the number of CT donors in 2020 was significantly lower than those of 2019 (66 vs. 99, p value 0.02). The total number of CT collected from donors have been 254 in 2019 and 206 in 2020 (p 0.28). Femoral arteries were the most required vascular tissues (55.5% in 2019 and 40% in 2020). Fifty-five and forty-eight pulmonary valves were implanted in 2019 and 2020, respectively. No differences were found for the types of CT requests between the 2 years. The median age of receivers of vascular tissues was 69.6 ± 14.6 years in the 2019 and 63.3 ± 14.9 years in 2020 (p < 0.01). The median age of receivers of pulmonary and aortic valves did not differ between the 2 years (9.32 ± 11.49 vs. 8.36 ± 10.66 and 48.67 ± 27.19 vs. 37.14 ± 31.97 respectively). Despite the dramatically reduction of donors, the number of CT collected has not decreased significantly and so far the CT distribution rate is comparable to those of 2019.

Aortic valve leaflet glycosaminoglycans composition and modification in severe chronic valve regurgitation.

BACKGROUND AND AIM OF THE STUDY: The surgical segments of aortic valve leaflets from patients with severe chronic aortic regurgitation were analyzed (by percentage and structure) for their content of complex polysaccharides and glycosaminoglycans (GAGs), and compared with control segments. METHODS: The GAG, hyaluronic acid (HA), chondroitin sulfate (CS) and dermatan sulfate (DS) and disaccharide contents were determined in segments (leaflet, root attachment region and belly) of aortic valve leaflets (non-coronary, left coronary and right coronary) using a multi-analytical approach. RESULTS: The aortic valve leaflets showed the presence of HA and CS/DS, with an overall charge density of -0.51-0.55. The CS/DS polymers showed a 4-sulfated/6-sulfated ratio of -0.70-0.77 in the belly, and -1.60-1.72 in commissure parts (-/+124%). The total amount of GAGs was -1.60-2.40 microg/mg of tissue. A significant increase in sulfated GAGs was observed in all valve parts in patients suffering from severe aortic insufficiency, as well as an increase in the 4-sulfated/6-sulfated ratio in the leaflet belly (-/+102%). CONCLUSION: It is speculated that differences in 4-sulfated/6-sulfated ratio determined in the belly and leaflet attachment region-commissure parts of the leaflets may correlate with the tensile or compressive loading of normal aortic valve regions. At the same time, it may be assumed that the increase in sulfated GAGs and 4-sulfated/6-sulfated ratio in the leaflet belly of valves taken from patients suffering severe aortic insufficiency was consistent with an altered matrix microstructure capable of influencing the hydration of these pathological tissues, and of conditioning their mechanical weakness.

Circulating microRNAs are new and sensitive biomarkers of myocardial infarction.

AIMS: Circulating microRNAs (miRNAs) may represent a novel class of biomarkers; therefore, we examined whether acute myocardial infarction (MI) modulates miRNAs plasma levels in humans and mice. METHODS AND RESULTS: Healthy donors (n = 17) and patients (n = 33) with acute ST-segment elevation MI (STEMI) were evaluated. In one cohort (n = 25), the first plasma sample was obtained 517 ± 309 min after the onset of MI symptoms and after coronary reperfusion with percutaneous coronary intervention (PCI); miR-1, -133a, -133b, and -499-5p were ~15- to 140-fold control, whereas miR-122 and -375 were ~87-90% lower than control; 5 days later, miR-1, -133a, -133b, -499-5p, and -375 were back to baseline, whereas miR-122 remained lower than control through Day 30. In additional patients (n = 8; four treated with thrombolysis and four with PCI), miRNAs and troponin I (TnI) were quantified simultaneously starting 156 ± 72 min after the onset of symptoms and at different times thereafter. Peak miR-1, -133a, and -133b expression and TnI level occurred at a similar time, whereas miR-499-5p exhibited a slower time course. In mice, miRNAs plasma levels and TnI were measured 15 min after coronary ligation and at different times thereafter. The behaviour of miR-1, -133a, -133b, and -499-5p was similar to STEMI patients; further, reciprocal changes in the expression levels of these miRNAs were found in cardiac tissue 3-6 h after coronary ligation. In contrast, miR-122 and -375 exhibited minor changes and no significant modulation. In mice with acute hind-limb ischaemia, there was no increase in the plasma level of the above miRNAs. CONCLUSION: Acute MI up-regulated miR-1, -133a, -133b, and -499-5p plasma levels, both in humans and mice, whereas miR-122 and -375 were lower than control only in STEMI patients. These miRNAs represent novel biomarkers of cardiac damage.

Neuropeptide Y promotes adipogenesis of human cardiac mesenchymal stromal cells in arrhythmogenic cardiomyopathy.

BACKGROUND: Arrhythmogenic Cardiomyopathy (AC) is a familial cardiac disease, mainly caused by mutations in desmosomal genes. AC hearts show fibro-fatty myocardial replacement, which favors stress-related life-threatening arrhythmias, predominantly in the young and athletes. AC lacks effective therapies, as its pathogenesis is poorly understood. Recently, we showed that cardiac Mesenchymal Stromal Cells (cMSCs) contribute to adipose tissue in human AC hearts, although the underlying mechanisms are still unclear. PURPOSE: We hypothesize that the sympathetic neurotransmitter, Neuropeptide Y (NPY), participates to cMSC adipogenesis in human AC. METHODS: For translation of our findings, we combined in vitro cytochemical, molecular and pharmacologic assays on human cMSCs, from myocardial biopsies of healthy controls and AC patients, with the use of existing drugs to interfere with the predicted AC mechanisms. Sympathetic innervation was inspected in human autoptic heart samples, and NPY plasma levels measured in healthy and AC subjects. RESULTS: AC cMSCs expressed higher levels of pro-adipogenic isotypes of NPY-receptors (i.e. Y1-R, Y5-R). Consistently, NPY enhanced adipogenesis in AC cMSCs, which was blocked by FDA-approved Y1-R and Y5-R antagonists. AC-associated PKP2 reduction directly caused NPY-dependent adipogenesis in cMSCs. In support of the involvement of sympathetic neurons (SNs) and NPY in AC myocardial remodeling, patients had elevated NPY plasma levels and, in human AC hearts, SNs accumulated in fatty areas and were close to cMSCs. CONCLUSIONS: Independently from the disease origin, AC causes in cMSCs a targetable gain of responsiveness to NPY, which leads to increased adipogenesis, thus playing a role in AC myocardial remodeling.

Human Cardiac Mesenchymal Stromal Cells From Right and Left Ventricles Display Differences in Number, Function, and Transcriptomic Profile.

BACKGROUND: Left ventricle (LV) and right ventricle (RV) are characterized by well-known physiological differences, mainly related to their different embryological origin, hemodynamic environment, function, structure, and cellular composition. Nevertheless, scarce information is available about cellular peculiarities between left and right ventricular chambers in physiological and pathological contexts. Cardiac mesenchymal stromal cells (C-MSC) are key cells affecting many functions of the heart. Differential features that distinguish LV from RV C-MSC are still underappreciated. AIM: To analyze the physiological differential amount, function, and transcriptome of human C-MSC in LV versus (vs.) RV. METHODS: Human cardiac specimens of LV and RV from healthy donors were used for tissue analysis of C-MSC number, and for C-MSC isolation. Paired LV and RV C-MSC were compared as for surface marker expression, cell proliferation/death ratio, migration, differentiation capabilities, and transcriptome profile. RESULTS: Histological analysis showed a greater percentage of C-MSC in RV vs. LV tissue. Moreover, a higher C-MSC amount was obtained from RV than from LV after isolation procedures. LV and RV C-MSC are characterized by a similar proportion of surface markers. Functional studies revealed comparable cell growth curves in cells from both ventricles. Conversely, LV C-MSC displayed a higher apoptosis rate and RV C-MSC were characterized by a higher migration speed and collagen deposition. Consistently, transcriptome analysis showed that genes related to apoptosis regulation or extracellular matrix organization and integrins were over-expressed in LV and RV, respectively. Besides, we revealed additional pathways specifically associated with LV or RV C-MSC, including energy metabolism, inflammatory response, cardiac conduction, and pluripotency. CONCLUSION: Taken together, these results contribute to the functional characterization of RV and LV C-MSC in physiological conditions. This information suggests a possible differential role of the stromal compartment in chamber-specific pathologic scenarios.

[Reducing the impact of COVID-19 on radiation oncology units of developing countries: A rapid review and expert consensus]. / Reduciendo el impacto de COVID-19 sobre la radioterapia oncológica en países en desarrollo: revisión rápida y consenso de expertos.

OBJECTIVE: To generate recommendations on the management of radiotherapeutic treatments during the pandemic, adapted to a country with limited health resources. METHODS: We did a rapid review of the literature, searching for papers that describe any measures to reduce the risk of COVID-19 infection, as well as management guidelines to reduce the workload, in radiotherapy units. The following conditions were included in the scope of this review: gynecological tumors, breast cancer, gastrointestinal tumors, genitourinary tumors, head and neck tumors, skin cancer, tumors of the central nervous system, and lymphomas. An expert group discussed online the extracted data and drafted the recommendations. Using a modified Delphi method, the consensus was reached among 14 certificated radio-oncologists. The quality of the evidence that supported the recommendations on treatment schedules was assessed. RESULTS: A total of 57 documents were included. Of these, 25 provided strategies to reduce the risk of infection. Recommendations for each condiction were extracted from the remaining documents. The recommendations aim to establish specific parameters where treatments can be omitted, deferred, prioritized, and shortened. Treatment schemes are recommended for each condition, prioritizing hypo-fractionated schemes whenever possible. CONCLUSIONS: We propose strategies for the management of radiotherapy services to guarantee the continuity of high-quality treatments despite the health crisis caused by COVID-19.

OBJETIVO: Establecer recomendaciones para la toma de decisiones de manejo en radioterapia durante la pandemia de COVID-19, adaptadas a un país con recursos de salud limitados. MÉTODOS: A través de una revisión rápida de la literatura se buscaron publicaciones que describieran medidas para reducir el riesgo de infección por COVID-19, así como también pautas de manejo para reducir la carga de trabajo en las unidades de radioterapia. Se incluyeron en el alcance de esta revisión las siguientes patologías: tumores ginecológicos, cáncer de mama, tumores gastrointestinales, tumores genitourinarios, tumores de cabeza y cuello, cáncer de piel, tumores del sistema nervioso central y linfomas. Un grupo de expertos discutió en línea los datos extraídos y redactó las recomendaciones. Mediante un método Delphi modificado, se evaluó el consenso entre 14 radio-oncólogos certificados. Se evaluó la calidad de la evidencia que sustentó las recomendaciones sobre esquemas de tratamiento. RESULTADOS: Se incluyeron un total de 57 documentos. De 25 trabajos se extrajeron las estrategias para reducir el riesgo de infección. De los restantes, se obtuvieron las recomendaciones para cada patología. Las recomendaciones están orientadas a establecer escenarios específicos donde se pueden omitir, diferir, priorizar y acortar los tratamientos. En el ítem de acortar se recomiendan esquemas de tratamiento para cada patología, priorizando los esquemas hipofraccionados cuando fue posible. CONCLUSIÓN: Se plantean estrategias para la gestión de los servicios de radioterapia con el objetivo de garantizar que los tratamientos de alta calidad para pacientes oncológicos sigan entregándose, pese a la crisis sanitaria ocasionada por COVID-19.

Manejo multidisciplinario de las metástasis cerebrales: una revisión actualizada y un cambio de paradigma / Multidisciplinary management of brain metastases: An updated review and paradigm shift

Introducción: Las metástasis cerebrales (MC) representan una problemática importante de la salud pública, en promedio el 30% de los pacientes oncológicos desarrollarán MC siendo importante causa de morbilidad, ansiedad y mortalidad. El tratamiento con radioterapia, cirugía y sistémico son los pilares para el tratamiento y han tenido una evolución importante en la última década. Propósito de la revisión: Se proporciona información actualizada en relación a epidemiología, diagnóstico, pronóstico y tratamiento de metástasis cerebrales desde un enfoque multidisciplinariopara lograr individualizar su abordaje con el objetivo de proporcionar control oncológico y calidad de vida. También se discuten el acceso a nuevas terapias sistémicas, técnicas quirúrgicas y disponibilidad de tecnología para ofrecer técnicas avanzadas de radioterapia.Men saje principal: Conocer mutaciones específicas y receptores diana de los tumores permite elegir quimio-inmunoterapia o terapias dirigidas actuales que ofrecen mejor potencial de control tanto a nivel sistémico como intracraneal. La secuenciación de los tratamientos sistémicos y locales (cirugía, radio-cirugía, radioterapia holocraneal) deben ser discutidos desde un enfoque multidisciplinario Conclusión: Es importante poder estimar el pronóstico de los pacientes con MC, esto determinarla conducta terapéutica que puede variar desde cuidados sintomáticos hasta tratamientos más agresivos como resección neuroquirúrgica o radiocirugía.

In troduction:Brain metastases (BMs) represent a significant public health problem. An average of 30% of cancer patients develop BM, which is a significant cause of morbidity, anxiety, and mortality. Radio-therapy, surgery, and systemic treatment are the mainstays of treatment and have evolved significantly in the last decade.Pu rpose of the review: Updated information on the epidemiology, diagnosis, prognosis, and treatment of brain metastases from a multidisciplinary approach is provided to enable an individualized approach aimed at cancer control and quality of life. Access to new systemic therapies, surgical techniques, and availability of technology for advanced radiotherapy techniques are also discussed.Mai n message: Knowledge of specific mutations and targets of tumor receptors allows the selection of chemoimmunotherapy or current targeted therapies that offer better control potential at the systemic and intracranial levels. The sequence of systemic and local treatments (surgery, radiosurgery, whole brain radiation therapy) should be discussed as part of a multidisciplinary approach.C o nclusion: It is essential to estimate the prognosis of patients with BM, given that this will determine the therapeuticbehavior that can range from symptomatic care to more aggressive treatments such as neurosurgical resection or radiosurgery

Radioterapia hipofraccionada para cáncer de próstata localizado ¿Menos tiempo de tratamiento es mejor? / Hypofractionated radiotherapy for localized prostate cancer ¿Is less treatment time better?

Introducción: El hipofraccionamiento moderado (hRt) en cáncer de próstata, consisten en dismi-nuir el tiempo total de tratamiento con radioterapia, lo que mejora la adherencia terapéutica y opti-miza recursos tecnológicos. En cáncer de próstata, existe evidencia robusta con datos maduros a 5 años de seguimiento, donde se evidenció que hRtno es inferior al tratamiento con fraccionamiento estándar en control oncológico, con menor o igual toxicidad aguda y tardía. Se hace una revisión de la evidencia, dosis de tolerancia, contorneo de volúmenes objetivo (GTV-CTV-PTV) / órganos de ries-go, planificación y reproducibilidad del hRt en cáncer de próstata localizado.

Introduction: Moderate hypofractionation (hRt) in prostate cancer consists of reducing the total time of treatment with radiotherapy, which improves therapeutic adherence and optimizes technolog-ical resources. In prostate cancer, there is robust evidence with mature data at 5 years of follow-up, where it was evidenced that hRt is not inferior to treatment with standard fractionation in oncological control, with less or equal acute and late toxicity. A review of the evidence, tolerance dose, contouring of target volumes (GTV-CTV-PTV) / organs at risk, planning and reproducibility of hRt in localized prostate cancer is made.

Reduciendo el impacto de COVID-19 sobre la radioterapia oncológica en países en desarrollo: revisión rápida y consenso de expertos / Reducing the impact of COVID-19 on radiation oncology units of developing countries: a rapid review and expert consensus

OBJETIVO: Establecer recomendaciones para la toma de decisiones de manejo en radioterapia durante la pandemia de COVID-19, adaptadas a un país con recursos de salud limitados. MÉTODOS: A través de una revisión rápida de la literatura se buscaron publicaciones que describieran medidas para reducir el riesgo de infección por COVID-19, así como también pautas de manejo para reducir la carga de trabajo en las unidades de radioterapia. Se incluyeron en el alcance de esta revisión las siguientes patologías: tumores ginecológicos, cáncer de mama, tumores gastrointestinales, tumores genitourinarios, tumores de cabeza y cuello, cáncer de piel, tumores del sistema nervioso central y linfomas. Un grupo de expertos discutió en línea los datos extraídos y redactó las recomendaciones. Mediante un método Delphi modificado, se evaluó el consenso entre 14 radio-oncólogos certificados. Se evaluó la calidad de la evidencia que sustentó las recomendaciones sobre esquemas de tratamiento. RESULTADOS: Se incluyeron un total de 57 documentos. De 25 trabajos se extrajeron las estrategias para reducir el riesgo de infección. De los restantes, se obtuvieron las recomendaciones para cada patología. Las recomendaciones están orientadas a establecer escenarios específicos donde se pueden omitir, diferir, priorizar y acortar los tratamientos. En el ítem de acortar se recomiendan esquemas de tratamiento para cada patología, priorizando los esquemas hipofraccionados cuando fue posible. CONCLUSIÓN: Se plantean estrategias para la gestión de los servicios de radioterapia con el objetivo de garantizar que los tratamientos de alta calidad para pacientes oncológicos sigan entregándose, pese a la crisis sanitaria ocasionada por COVID-19.

OBJECTIVE: To generate recommendations on the management of radiotherapeutic treatments during the pandemic, adapted to a country with limited health resources. METHODS: We did a rapid review of the literature, searching for papers that describe any measures to reduce the risk of COVID-19 infection, as well as management guidelines to reduce the workload, in radiotherapy units. The following conditions were included in the scope of this review: gynecological tumors, breast cancer, gastrointestinal tumors, genitourinary tumors, head and neck tumors, skin cancer, tumors of the central nervous system, and lymphomas. An expert group discussed online the extracted data and drafted the recommendations. Using a modified Delphi method, the consensus was reached among 14 certificated radio-oncologists. The quality of the evidence that supported the recommendations on treatment schedules was assessed. RESULTS: A total of 57 documents were included. Of these, 25 provided strategies to reduce the risk of infection. Recommendations for each condiction were extracted from the remaining documents. The recommendations aim to establish specific parameters where treatments can be omitted, deferred, prioritized, and shortened. Treatment schemes are recommended for each condition, prioritizing hypo-fractionated schemes whenever possible. CONCLUSIONS: We propose strategies for the management of radiotherapy services to guarantee the continuity of high-quality treatments despite the health crisis caused by COVID-19.

Mechanical compliance and immunological compatibility of fixative-free decellularized/cryopreserved human pericardium.

BACKGROUND: The pericardial tissue is commonly used to produce bio-prosthetic cardiac valves and patches in cardiac surgery. The procedures adopted to prepare this tissue consist in treatment with aldehydes, which do not prevent post-graft tissue calcification due to incomplete xeno-antigens removal. The adoption of fixative-free decellularization protocols has been therefore suggested to overcome this limitation. Although promising, the decellularized pericardium has not yet used in clinics, due to the absence of proofs indicating that the decellularization and cryopreservation procedures can effectively preserve the mechanical properties and the immunologic compatibility of the tissue. PRINCIPAL FINDINGS: The aim of the present work was to validate a procedure to prepare decellularized/cryopreserved human pericardium which may be implemented into cardiovascular homograft tissue Banks. The method employed to decellularize the tissue completely removed the cells without affecting ECM structure; furthermore, uniaxial tensile loading tests revealed an equivalent resistance of the decellularized tissue to strain, before and after the cryopreservation, in comparison with the fresh tissue. Finally, immunological compatibility, showed a minimized host immune cells invasion and low levels of systemic inflammation, as assessed by tissue transplantation into immune-competent mice. CONCLUSIONS: Our results indicate, for the first time, that fixative-free decellularized pericardium from cadaveric tissue donors can be banked according to Tissue Repository-approved procedures without compromising its mechanical properties and immunological tolerance. This tissue can be therefore treated as a safe homograft for cardiac surgery.

Metástasis endometrial por cáncer de mama durante hormonoterapia adyuvante: reporte de caso / Endometrial metastasis for breast cancer during adjuvant hormone therapy: case report

Introducción: el cáncer de mama es el tumor más frecuente en las mujeres y la primera causa de muerte en este grupo con cerca de 230.000 casos nuevos al año en los Estados Unidos. En el tratamiento adyuvante del cáncer de mama con receptores hormonales positivos el tamoxifeno cumple un papel fundamental sin embargo es bien conocido que su uso incrementa el riesgo de cáncer endometrial. Las metástasis del cáncer de mama hacia el útero son poco comunes aunque existen algunos casos reportados en la literatura. El diagnóstico diferencial entre tumor primario y metástasis distantes de cáncer de mama hacia el útero determina el tipo de tratamiento que se deberá realizar. Caso: nosotros reportamos un caso de cáncer de mama que se presenta con metástasis uterina durante el tratamiento con tamoxifeno y demostramos la utilidad del estudio histopatológico con inmunohistoquimica para llegar al diagnóstico.

Introduction: breast cancer is the most common tumor in women and the leading cause of cancer death in this group with about 230,000 new cases per year in the United States. Adjuvant treatment of breast cancer with hormone receptor-positive tamoxifen plays a critical role but is nevertheless well known the increased risk of endometrial cancer with its use. Metastases of breast cancer to the uterus are rare but there are some cases reported in the literature. The differential diagnosis between primary tumor and distant metastases to the uterus from breast cancer determines the type of treatment that should be administered. Case: we report a case of breast cancer that presented uterine metastases during treatment with tamoxifen and demonstrate the usefulness of histopathology with immunohistochemistry to reach the diagnosis.

[Cardiac engineered valves: state of the art]. / Valvole cardiache ingegnerizzate: stato dell'arte.

Engineered heart valves promise to be the ideal heart valve replacement: they have the potential to grow and repair within the host, to minimize inflammatory and immunological responses and to limit thromboembolism. In particular, pediatric patients with congenital defects would greatly benefit from growing replacement materials by reducing the need for surgical reinterventions due to the lack of prosthetic material growth. In fact, viable cells included in engineered heart valves can theoretically adapt to a growing and changing environment exactly as a native biological structure. Engineered heart valves represent thus a novel experimental approach to create autologous, living replacements with the potential to grow, regenerate, and remodel. The aim of this paper is to present a review concerning the research efforts to create a viable engineered heart valve.

Rates of cycling cells in cryopreserved valvular homograft: a preliminary study.

Some investigators claim that the viability of cryopreserved human valvular homograft is necessary for the duration of implanted homograft. In this preliminary study, the percentage of cycling cells in cryopreserved valvular homografts was evaluated with the use of monoclonal Ki-67 antibody. Three human aortic valves were harvested from multiorgan donors and cryopreserved. Sections of 5 microm in thickness were stained with monoclonal Ki-67 antibody. The proportion of endothelial cells with Ki-67 positive nuclei was 1.80 +/- 0.20%. No differences in distribution were observed from basal to marginal sites. Few fibroblasts showed Ki-67-immunopositivity (0.10 +/- 0.06%) while the Ki-67 immunostaining was 0.80 +/- 0.20% in myocytes. Our preliminary study shows that cryopreserved valvular homograft cells are not only viable but they also have the potential to replicate. These data can lead to the hypothesis that valvular cells could actively replicate even after implantation, permitting cellular renewal and regeneration of extracellular matrix's components.

Effect of cryopreservation techniques on aortic valve glycosaminoglycans.

This study was designed to evaluate the effect of cryopreservation on the glycosaminoglycan (GAG) content of the aortic allografts. Twenty-one porcine aortic valves were obtained. Five aortic roots were immediately analyzed without cryopreservation, eight were cryopreserved in closed leaflet position, and eight in open leaflet position. The groups were compared in terms of GAG concentration and subclass proportion in three different zones including the aortic root wall, the commissures, and the leaflets. GAG content at the commissures was significantly lower in the closed leaflet group than in the other groups (P = 0.001). The electrophoretic analysis did not show any significant difference in the zonal distribution of GAG classes between groups. Quantitative analysis in various aortic valve zones suggests that cryopreservation can alter the GAG content. Cryopreservation of the aortic valve in an open leaflet position can preserve the matrix more efficiently and might prolong the durability of the aortic allograft.