Live Organoid Cyclic Imaging.

Organoids are becoming increasingly relevant in biology and medicine for their physiological complexity and accuracy in modeling human disease. To fully assess their biological profile while preserving their spatial information, spatiotemporal imaging tools are warranted. While previously developed imaging techniques, such as four-dimensional (4D) live imaging and light-sheet imaging have yielded important clinical insights, these technologies lack the combination of cyclic and multiplexed analysis. To address these challenges, bioorthogonal click chemistry is applied to display the first demonstration of multiplexed cyclic imaging of live and fixed patient-derived glioblastoma tumor organoids. This technology exploits bioorthogonal click chemistry to quench fluorescent signals from the surface and intracellular of labeled cells across multiple cycles, allowing for more accurate and efficient molecular profiling of their complex phenotypes. Herein, the versatility of this technology is demonstrated for the screening of glioblastoma markers in patient-derived human glioblastoma organoids while conserving their viability. It is anticipated that the findings and applications of this work can be broadly translated into investigating physiological developments in other organoid systems.

Genomic consequences of isolation and inbreeding in an island dingo population.

Dingoes come from an ancient canid lineage that originated in East Asia around 8000-11,000 years BP. As Australia's largest terrestrial predator, dingoes play an important ecological role. A small, protected population exists on a world heritage listed offshore island, K'gari (formerly Fraser Island). Concern regarding the persistence of dingoes on K'gari has risen due to their low genetic diversity and elevated inbreeding levels. However, whole-genome sequencing data is lacking from this population. Here, we include five new whole-genome sequences of K'gari dingoes. We analyze a total of 18 whole genome sequences of dingoes sampled from mainland Australia and K'gari to assess the genomic consequences of their demographic histories. Long (>1 Mb) runs of homozygosity (ROH) - indicators of inbreeding - are elevated in all sampled dingoes. However, K'gari dingoes showed significantly higher levels of very long ROH (>5 Mb), providing genomic evidence for small population size, isolation, inbreeding, and a strong founder effect. Our results suggest that, despite current levels of inbreeding, the K'gari population is purging strongly deleterious mutations, which, in the absence of further reductions in population size, may facilitate the persistence of small populations despite low genetic diversity and isolation. However, there may be little to no purging of mildly deleterious alleles, which may have important long-term consequences, and should be considered by conservation and management programs.

CD39+ conventional CD4+ T cells with exhaustion traits and cytotoxic potential infiltrate tumors and expand upon CTLA-4 blockade.

Conventional CD4+ T (Tconv) lymphocytes play important roles in tumor immunity; however, their contribution to tumor elimination remains poorly understood. Here, we describe a subset of tumor-infiltrating Tconv cells characterized by the expression of CD39. In several mouse cancer models, we observed that CD39+ Tconv cells accumulated in tumors but were absent in lymphoid organs. Compared to tumor CD39- counterparts, CD39+ Tconv cells exhibited a cytotoxic and exhausted signature at the transcriptomic level, confirmed by high protein expression of inhibitory receptors and transcription factors related to the exhaustion. Additionally, CD39+ Tconv cells showed increased production of IFNγ, granzyme B, perforin and CD107a expression, but reduced production of TNF. Around 55% of OVA-specific Tconv from B16-OVA tumor-bearing mice, expressed CD39. In vivo CTLA-4 blockade induced the expansion of tumor CD39+ Tconv cells, which maintained their cytotoxic and exhausted features. In breast cancer patients, CD39+ Tconv cells were found in tumors and in metastatic lymph nodes but were less frequent in adjacent non-tumoral mammary tissue and not detected in non-metastatic lymph nodes and blood. Human tumor CD39+ Tconv cells constituted a heterogeneous cell population with features of exhaustion, high expression of inhibitory receptors and CD107a. We found that high CD4 and ENTPD1 (CD39) gene expression in human tumor tissues correlated with a higher overall survival rate in breast cancer patients. Our results identify CD39 as a biomarker of Tconv cells, with characteristics of both exhaustion and cytotoxic potential, and indicate CD39+ Tconv cells as players within the immune response against tumors.

Clinical characteristics of patients with Breast and / or Ovarian Cancer with mutations in the BRCA1 and BRCA2 genes in Córdoba, Argentina / [Características clínicas de pacientes con Cáncer de Mama y / o Cáncer de Ovario con mutaciones en los genes BRCA1 y BRCA2 en Córdoba, Argentina.]

Introduction: Hereditary predisposition syndromes to cancer represent 5-10% of cancer cases, the most studied being HBOC produced by mutations in the BRCA1/2 genes. Objectives: To describe clinical, histopathological and PV characteristics in patients with HBOC in Córdoba, Argentina and compare it with those without BRCA1/2 mutations. Methods: Cross-sectional, correlational and observational analysis of patients from Córdoba. The ANOVA, Student's t test contingency tables and Fisher exact test were used the significance level was α = 0.05. Results: 155 women with BC, OC and BC/OC were studied. 40 BRCA1 / 2 mutations were identified. No differences were found in the age of diagnosis between patients with and without BRCA1/2 mutations. A significant association was found between VP in BRCA1/2 and the type of cancer (p = 0.003); all cases with BC/OC presented mutations in BRCA1/2. No significant association was found between mutated/non-mutated and personal history, family background, and ER-PR-HER2. 23.1% and 38.1% of BC cases were TN in individuals with VP in BRCA 1 and 2, respectively. The prevalence of mutations was 25.8% and the prevalence of novel PV was 10.0%. Conclusions: Patients with BC-VP BRCA1/2 are associated with ductal histology, and younger age of presentation with VP BRCA1. We did not find significant differences in the age at diagnosis of BC between patients with BRCA1 and BRCA2 mutations, a higher proportion of BC TN is observed than in the general population. In our sample, the prevalence of BRCA1/2 mutations among patients who meet criteria for HBOC is 25.8%, with 10% new pathogenic variant.

Introducción: Los síndromes de predisposición hereditaria al cáncer representan un 5-10% de los casos de cáncer, el más estudiado es HBOC producido por mutaciones en los genes BRCA1/2. Objetivos: Describir características clínicas, histopatológicas y VP en pacientes con HBOC en Córdoba, Argentina y compararla con aquellas sin mutaciones en BRCA1/2. Métodos: Análisis transversal, correlacional y observacional de pacientes de Córdoba. Se utilizó la prueba ANOVA, t de Student, tablas de contingencia y prueba exacta de Fisher, el nivel de significancia fue α=0,05. Resultados: Se estudiaron 155 mujeres con CM, CO y CM/CO. Se identificaron 40 mutaciones en BRCA1/2. No se encontraron diferencias en edad de diagnóstico entre pacientes con y sin mutaciones en BRCA1/2. Se encontró asociación significativa entre VP en BRCA1/2 y el tipo de cáncer (p=0,003); todos los casos con CM/CO presentaron mutaciones en BRCA1/2. No se encontró asociación significativa entre mutados/no mutados y AP, AF, RE-RP-HER2. El 23.1% y 38.1% de los casos de CM fueron TN en individuos con VP en BRCA 1 y 2 respectivamente. La prevalencia de mutaciones fue 25,8% y la prevalencia de VP noveles del 10,0%. Conclusiones: Las pacientes con CM-VP BRCA1/2 están asociadas con histología ductal, y menor edad de presentación con VP BRCA1. No encontramos diferencias significativas en edad de diagnóstico del CM entre pacientes con mutaciones BRCA1 y BRCA2, se observa una mayor proporción CM TN que en la población en general. En nuestra muestra, la prevalencia de mutaciones en BRCA1/2 entre los pacientes que reúnen criterios para HBOC es del 25,8%, con 10% de VP noveles.

Intensified fish farming. Performance of electrochemical remediation of marine RAS waters.

Aquaculture has been the fastest growing agricultural sector in the past few decades and currently supplies about half of the fish market. A range of environmental and management concerns including limited land and water availability have led to intensifying fish production by recirculating aquaculture systems (RAS). Fish's diet contains 30-60 % protein and about 4-10 % nitrogen (N). As fish assimilate only 20-30 % of the feed to produce body mass, the unassimilated N is released in the form of toxic ammonium that deteriorates water quality and compels its degradation. Widely extended biological nitrification is not efficient in the removal of nitrites nor other chemicals and pharmaceuticals used during fish culture. Electrochemical oxidation, a less developed alternative, reports several advantages such as, i) simultaneous degradation of ammonia­nitrogen (TAN) and water disinfection in the same step with considerable simplification of the whole process, ii) easy adaptability to different production scales and periods of fish growth, and iii) no generation of harmful by-products and no use of chemicals, among others. Besides, in the case of marine aquaculture, the technology benefits from the high conductivity of seawater; thus, electrochemical oxidation is positioned in a very good place to satisfy the water treatment needs of the increasing production rate of marine aquaculture fish. Here, we report the analysis of the performance of a RAS demonstration plant aimed at farming gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) and provided with electrochemical remediation of culture water. The performance of the plant, with 20 m3 of seawater operating at a recirculation rate of 0.9-1.4 h-1, has been analysed in terms of TAN removal, water disinfection, make-up water intake and energy consumption and compared to data of conventional RAS provided with biofilters. The benefits and advantages of the innovative electrochemical remediation of RAS water are highlighted.

Factores Predictivos de Compromiso Axilar en ocasión de Ganglio Centinela Positivo en Cáncer de Mama Temprano / Predictive Factors of Axillary Compromise on the Occasion of Positive Sentinel Lymph Node in Early Breast Cancer

Introducción: Se ha demostrado en diversos protocolos que, en gran proporción de casos con cáncer de mama en estadíos tempranos y ganglio centinela positivo, no se observa enfermedad metastásica en ganglios linfáticos no centinelas (GNC). A raíz de esto es que se han descripto diferentes factores predictores de metástasis axilar. Esto nos motivó a realizar el presente trabajo. Objetivo: Analizar diferentes factores anatomopatológicos y su influencia en el compromiso metastásico de los ganglios no centinela. Material y método: Se estudiaron casos de cáncer de mama estadíos tempranos (T1-T2) con ganglio centinela positivo y seguidas de vaciamiento axilar, operadas en IMGO entre febrero del año 2000 y diciembre de 2012. De la muestra total se identificaron dos grupos: GnC negativos y GnC positivos. Analizamos diferentes variables anatomopatológicas vinculadas a cada grupo y calculamos su grado de significancia. Finalmente, analizamos estas variables en una tabla de análisis multivariado. Resultados: De una muestra de 205 casos (20,8% del total de ganglios estudia- dos en ese período de tiempo), el 48.3% (n=99) presentaba metástasis solo en el GC. De todas las características anatomopatológicas de mal pronóstico analizadas encontramos que el tamaño tumoral 2, el subtipo lobulillar y la invasión linfovascular tuvieron correlación significativa con la cantidad de GnC comprometidos. En el análisis multivariado encontramos tres factores predictivos, la invasión linfovascular, el tamaño de la metástasis en el ganglio centinela y el número de ganglios centinelas comprometidos. Conclusiones: Los factores independientes de compromiso de ganglios no centinela (GNC) en nuestra serie fueron: la invasión linfovascular, el tamaño de la metástasis en el ganglio centinela y el número de ganglios centinelas comprometidos

Introduction: It has been shown in various protocols that pathologically negative nonsentinel lymph node (NSN) disease, is observed in the rest of the armpit in a large proportion of cases with early stage breast carcinoma and positive sentinel node. As a result of this, different pathological predictors of axillary metastasis have been described. This motivated us to carry out the present work. Objetive: To analyze different pathological factors and their influence on the metastatic involvement of nonsentinel nodes. Material and method: Early stage breast cancer cases (T1-T2) with positive sentinel node and axillary emptying, operated on in IMGO between February 2000 and December 2012, were studied. Two groups were identified from the total sample: negative GnC and positive GnC, and analyzed diffe- rent pathological variables associated with metastatic nonsentinel nodes, calculating their significance. Finally, we analyze these variables in a multivariate analysis table. Results: From a sample of 205 cases (20.8% of the total lymph nodes studied), 48.3% (n = 99) had metastasis only in the SN. Of all the anatomo-pathological characteristics analyzed, we found that tumor size two, lobular subtype, and lymphovascular invasion had a significant correlation with the amount of compromised non-sentinel node (NSN). In the multivariate analysis, we found three predictive factors: lymphovascular invasion, the size of the metastasis in the sentinel node and the number of positive sentinel nodes. Conclusions: The independent factors of nonsentinel node involvement (NSN) in our series are: lymphovascular invasion, the size of the sentinel node metastasis, and the number of sentinel nodes involved

Polyfunctional KLRG-1+CD57+ Senescent CD4+ T Cells Infiltrate Tumors and Are Expanded in Peripheral Blood From Breast Cancer Patients.

Senescent T cells have been described during aging, chronic infections, and cancer; however, a comprehensive study of the phenotype, function, and transcriptional program of this T cell population in breast cancer (BC) patients is missing. Compared to healthy donors (HDs), BC patients exhibit an accumulation of KLRG-1+CD57+ CD4+ and CD8+ T cells in peripheral blood. These T cells infiltrate tumors and tumor-draining lymph nodes. KLRG-1+CD57+ CD4+ and CD8+ T cells from BC patients and HDs exhibit features of senescence, and despite their inhibitory receptor expression, they produce more effector cytokines and exhibit higher expression of Perforin, Granzyme B, and CD107a than non-senescent subsets. When compared to blood counterparts, tumor-infiltrating senescent CD4+ T cells show similar surface phenotype but reduced cytokine production. Transcriptional profiling of senescent CD4+ T cells from the peripheral blood of BC patients reveals enrichment in genes associated with NK or CD8+-mediated cytotoxicity, TCR-mediated stimulation, and cell exhaustion compared to non-senescent T cells. Comparison of the transcriptional profile of senescent CD4+ T cells from peripheral blood of BC patients with those of HDs highlighted marked similarities but also relevant differences. Senescent CD4+ T cells from BC patients show enrichment in T-cell signaling, processes involved in DNA replication, p53 pathways, oncogene-induced senescence, among others compared to their counterparts in HDs. High gene expression of CD4, KLRG-1, and B3GAT1 (CD57), which correlates with increased overall survival for BC patients, underscores the usefulness of the evaluation of the frequency of senescent CD4+ T cells as a biomarker in the follow-up of patients.

Single-cell barcode analysis provides a rapid readout of cellular signaling pathways in clinical specimens.

Serial tissue sampling has become essential in guiding modern targeted and personalized cancer treatments. An alternative to image guided core biopsies are fine needle aspirates (FNA) that yield cells rather than tissues but are much better tolerated and have lower complication rates. The efficient pathway analysis of such cells in the clinic has been difficult, time consuming and costly. Here we develop an antibody-DNA barcoding approach where harvested cells can be rapidly re-stained through the use of custom designed oligonucleotide-fluorophore conjugates. We show that this approach can be used to interrogate drug-relevant pathways in scant clinical samples. Using the PI3K/PTEN/CDK4/6 pathways in breast cancer as an example, we demonstrate how analysis can be performed in tandem with trial enrollment and can evaluate downstream signaling following therapeutic inhibition. This approach should allow more widespread use of scant single cell material in clinical samples.

CD39 Expression Defines Cell Exhaustion in Tumor-Infiltrating CD8+ T Cells.

The ability of CD8+ T lymphocytes to eliminate tumors is limited by their ability to engender an immunosuppressive microenvironment. Here we describe a subset of tumor-infiltrating CD8+ T cells marked by high expression of the immunosuppressive ATP ecto-nucleotidase CD39. The frequency of CD39highCD8+ T cells increased with tumor growth but was absent in lymphoid organs. Tumor-infiltrating CD8+ T cells with high CD39 expression exhibited features of exhaustion, such as reduced production of TNF and IL2 and expression of coinhibitory receptors. Exhausted CD39+CD8+ T cells from mice hydrolyzed extracellular ATP, confirming that CD39 is enzymatically active. Furthermore, exhausted CD39+CD8+ T cells inhibited IFNγ production by responder CD8+ T cells. In specimens from breast cancer and melanoma patients, CD39+CD8+ T cells were present within tumors and invaded or metastatic lymph nodes, but were barely detectable within noninvaded lymph nodes and absent in peripheral blood. These cells exhibited an exhausted phenotype with impaired production of IFNγ, TNF, IL2, and high expression of coinhibitory receptors. Although T-cell receptor engagement was sufficient to induce CD39 on human CD8+ T cells, exposure to IL6 and IL27 promoted CD39 expression on stimulated CD8+ T cells from human or murine sources. Our findings show how the tumor microenvironment drives the acquisition of CD39 as an immune regulatory molecule on CD8+ T cells, with implications for defining a biomarker of T-cell dysfunction and a target for immunotherapeutic intervention.Significance: The tumor microenvironment elicits a subset of functionally exhausted CD8+ T cells by creating conditions that induce cell surface expression of CD39, an immunosuppressive molecule that can be therapeutically targeted to restore effector T-cell function. Cancer Res; 78(1); 115-28. ©2017 AACR.

Irradiación parcial acelerada de mama (apbi) después de cuadrantectomía más biopsia del ganglio centinela axilar: características clínico-patológicas e inmunohistoquímicas / Accelerated partial breast irradiation (apbi) after breast conservative surgery plus sentinel node biopsy: clinical-pathological and immunohistochemical characteristics

Introducción El tratamiento estándar en estadios tempranos del cáncer de mama es la cirugía conservadora (cc) más radioterapia del volumen total mamario (wbi). Dado que la mayoría de las recidivas locales ocurren cercanas al sitio del tumor (85%) y existiendo la posibilidad de acortar tiempos de irradiación ­debido a que esta insume varias semanas­, podría no ser necesario irradiar toda la mama, sino el área donde estaba situado el tumor más margen de seguridad, lo que insume un menor tiempo. Este el fundamento de la Irradiación Parcial Acelerada Mama (apbi). Consensos internacionales proporcionan guías para administrar apbi fuera de ensayos clínicos. Objetivos El objetivo principal de este trabajo es determinar y analizar las características clinico-anatomopatológicas e inmunohistoquímicas de las pacientes que fueron operadas y luego seleccionadas para ser irradiadas con técnica de apbi con radioterapia externa con intensidad modulada guiada por imágenes (imrt). Material y método Se trata de un trabajo prospectivo, no randomizado. La muestra está constituida por las primeras 20 pacientes ingresadas al protocolo apbi. El reclutamiento se realizó entre marzo de 2012 y julio de 2014. Se analizaron las características clínico-patológicas e inmunohistoquímicas de esas pacientes luego de cc más biopsia del ganglio centinela (bgc). Resultados Media de seguimiento en meses: 37,7 (r: 22-51); edad media: 65,7 años. Tamaño tumoral medio por anatomía patológica: 12,03 mm. Media de número de ganglios centinelas extirpados: 1,7. Todos los tumores fueron unifocales. Los márgenes quirúrgicos resultaron negativos en todos los casos. Tumores Luminal A: 19 pacientes; tumores Luminal B: 1 paciente. Hormonoterapia adyuvante en todas las pacientes. Control locorregional: 100%. Conclusiones A pesar del corto tiempo de seguimiento y del escaso número de pacientes, este análisis sugiere que la apbi en el lecho del tumor marcado intraoperatoriamente con fiduciales de titanio es factible para pacientes que reúnen los criterios estrictos clínico-patológicos e inmunohistoquímicos acordes con las guías internacionales.

Introduction The standard conservative treatment for early stage breast carcinomas is the breast conservative surgery plus whole breast irradiation. Because most local recurrences occur close to the site of the primary tumor (80- 90%) and the possibility of shortening the irradiation times ­because it takes several weeks­, it may not be necessary to irradiate the entire breast, but the area where the tumor was located plus a margin of safety in less time. This is the basis of Accelerated Partial Irradiation of the Breast Objectives Determination and analysis of clinical-pathological and immunohistochemical characteristics in patients selected for Accelerated Partial Breast Irradiation (apbi) performed by imrt + igrt technique after breast conservative surgery plus sentinel node biopsy. Materials and method A prospective, non-randomized study of the first 20 patients performed apbi strictly following the recommendations of international consensus. It was initiated in March 2012 until July 2014. Clinical-pathological and immunohistochemical characteristics of these patients were analyzed to be selected for apbi, after breast conservative surgery plus sentinel node biopsy. Results Median follow up: 37.7 months (r: 22-51); average age: 65.7 years. Mean tumor size by pathological analysis: 12.03 mm. Mean number of sentinel nodes removed: 1.7. All tumors were unifocal. Negative surgical margins in all cases. Luminal A tumors: 19 patients; Luminal B tumors: 1 patient. Adjuvant hormone therapy in all patients. Loco-regional control: 100%. Conclusions Despite the short time of follow up and the small numbers of patients, this analysis suggests that apbi performed by imrt + igrt technique added to the placement of fiduciary marks at the time of surgery is feasible for highly selected patients who meet the clinical-pathological and immunohistochemical selection criteria according to international guidelines

Evaluación de la técnica de Biopsia de Ganglio Centinela Axilar a través de la incisión mamaria en la cirugía conservadora / Evaluation of Sentinel Lymph Node Axilar Biopsy through the mammary incision in conservative surgery

Introducción El abordaje del ganglio centinela por incisión mamaria única permite acceder al ganglio evitando la realización de incisión axilar, lo que disminuiría el linfedema, el dolor y las disestesias y mejoraría el resultado estético. Objetivos Describir el abordaje quirúrgico del ganglio centinela a través de la incisión mamaria utilizada para la tumorectomía del carcinoma mamario. Material y método Entre septiembre de 2013 y agosto de 2015, se evaluaron 83 pacientes consecutivas con axila clínicamente negativa, que fueron sometidas a cirugía conservadora más biopsia de ganglio centinela, un grupo mediante abordaje clásico (incisión axilar) y otro por la incisión mamaria. Se estudió: edad; tamaño tumoral y ubicación; tipo histológico; técnica de identificación del ganglio centinela; número de ganglios resecados; ganglios centinelas positivos; y complicaciones perioperatorias. Resultados La detección del ganglio centinela por una única incisión se logró en todas las pacientes (100% de éxito). La realización de una única incisión presentó menores complicaciones en comparación con las pacientes a las que se les realizó diferentes incisiones; además, se encontraron mejores resultados estéticos. Conclusiones El abordaje del ganglio centinela mediante la misma incisión de la cirugía mamaria conservadora presenta una excelente tasa de detección, ofreciendo menos complicaciones y mejores resultados estéticos.

Introduction The sentinel node approach through a single breast incision allows its detection avoiding axillary incision, which could reduce complications and improve aesthetic result. Objectives The aim of this work was to describe the surgical approach of sentinel node via the single incision used for conservative surgery for breast carcinoma. Materials and method Patients (n=83) with clinically negative axilla, who went through conservative surgery and axillary sentinel node biopsy using classical approach (axillary incision) and single breast incision, in early-stage breast cancer, were studied. Clinical, anatomical and pathological variables and perioperative complications were considered. Results Our results showed that the method of detection of sentinel node by a single breast incision was successful in 100 % of the cases. The performance of a single incision presented minor complications compared to patients who underwent different incisions; also better aesthetic results were also found. Conclusions The surgical approach of the sentinel node through the single incision of breast conserving surgery presents an excellent detection rate, fewer complications and better aesthetic results.

Multiparametric plasma EV profiling facilitates diagnosis of pancreatic malignancy.

Pancreatic ductal adenocarcinoma (PDAC) is usually detected late in the disease process. Clinical workup through imaging and tissue biopsies is often complex and expensive due to a paucity of reliable biomarkers. We used an advanced multiplexed plasmonic assay to analyze circulating tumor-derived extracellular vesicles (tEVs) in more than 100 clinical populations. Using EV-based protein marker profiling, we identified a signature of five markers (PDACEV signature) for PDAC detection. In our prospective cohort, the accuracy for the PDACEV signature was 84% [95% confidence interval (CI), 69 to 93%] but only 63 to 72% for single-marker screening. One of the best markers, GPC1 alone, had a sensitivity of 82% (CI, 60 to 95%) and a specificity of 52% (CI, 30 to 74%), whereas the PDACEV signature showed a sensitivity of 86% (CI, 65 to 97%) and a specificity of 81% (CI, 58 to 95%). The PDACEV signature of tEVs offered higher sensitivity, specificity, and accuracy than the existing serum marker (CA 19-9) or single-tEV marker analyses. This approach should improve the diagnosis of pancreatic cancer.

The clinical relevance of micropapillary carcinoma of the breast: a case-control study.

AIMS: To ascertain the prognostic relevance of micropapillary carcinoma, a specific type of breast tumour. METHODS AND RESULTS: We interrogated the clinical records of a series of 49 pure micropapillary carcinoma patients and 13 487 invasive ductal carcinoma patients, diagnosed and treated consecutively in our institution over a 9-year time-frame. Compared with invasive ductal carcinoma, patients with micropapillary carcinoma more frequently had moderately differentiated tumours (P = 0.02) with extensive peritumoral vascular invasion (P < 0.0001), associated with a significantly higher rate of axillary lymph node involvement (P < 0.0001). Survival data obtained by comparing 49 micropapillary carcinoma patients with a set of 98 invasive ductal carcinoma patients matched for age, tumour size and grade, peritumoral vascular invasion, immunohistochemically defined molecular subtype, number of positive lymph nodes and year of surgery showed that the micropapillary histotype did not add any independent information to the risk of locoregional (P = 0.48) or distant (P = 0.79) relapse, or overall survival (P = 0.60). CONCLUSIONS: Our data reinforce the notion that micropapillary carcinoma usually arises as a locally advanced disease, and provide evidence that micropapillary histology does not add any additional information on clinical outcome independent of clinicopathological characteristics such as lymph node status and immunohistochemically defined molecular subtype.

Repeating conservative surgery after ipsilateral breast tumor reappearance: criteria for selecting the best candidates.

BACKGROUND: Mastectomy is still considered the treatment of first choice in patients with ipsilateral breast tumor recurrence (IBTR) after breast-conserving surgery (BCS) and whole-breast radiotherapy. METHODS: We retrospectively evaluated 161 patients with invasive IBTR who underwent a second BCS in order to describe prognosis, determine predictive factors of outcome, and select the subset of patients with the best local control. Median follow-up after IBTR was 81 months. RESULTS: Median age at IBTR was 53 years. Five-year overall survival after IBTR was 84 % (95 % confidence interval [CI] 78-89). Five-year cumulative incidence of a second local event after IBTR was 29 % (95 % CI 22-37). At the multivariate analysis, IBTR size >2 cm and time to relapse ≤48 months significantly increased the risk of local reappearance (hazard ratio [HR] 3.3, 95 % CI 1.6-7.0; and HR 1.9, 95 % CI 1.1-3.5). The 5-year cumulative incidence of a further local reappearance of the tumor after repeating BCS was 15.2 % in the patients with IBTR ≤2 cm and time to IBTR >48 months, 31.2 % in the patients with IBTR ≤2 cm and time to IBTR ≤48 months, and 71.2 % in patients with IBTR >2 cm (P < 0.001). CONCLUSIONS: The best candidates for a second BCS are those with small (≤2 cm) and late (>48 months) IBTR. The information about the risk of a further local reappearance after repeating BCS should be shared with the patients in the decision making process.

Accelerated partial breast irradiation in early breast cancer: focus on intraoperative treatment with electrons (ELIOT).

Wide tumor resection plus postoperative whole breast irradiation is standard treatment for early breast cancer. Irradiation decreases recurrence rates, but may cause poor cosmesis, breast pain, and cardiac and lung toxicity. Accelerated partial breast irradiation is increasingly used in the hope of increasing convenience, decreasing sequelae and maintaining cure rates. Intraoperative radiotherapy with electrons is an attractive accelerated partial breast irradiation technique because collimator placement is under the direct control of the surgeon who removes the tumor, the skin is spared, shielding protects the chest wall and complete irradiation can be given in a single intraoperative session (avoiding 5-7 weeks of whole breast irradiation). Intraoperative radiotherapy with electrons seems as safe as whole breast irradiation; however, long-term results on local control and survival are not available yet.

Sentinel lymph node biopsy in multicentric breast cancer: five-year results in a large series from a single institution.

PURPOSE: This study was designed to present the 5-year results of patients with multicentric breast cancer who underwent sentinel lymph node biopsy (SLNB) in a single institution. METHODS: Between June 1999 and December 2007, 337 patients with multicentric breast cancer and a clinically negative axilla underwent lymphatic mapping by a single periareolar/peritumoral (n = 306) or a double peritumoral or subdermal injection (n = 31) of (99m)Tc-HSA nanocolloids. The sentinel lymph node (SLN) was evaluated by intraoperative frozen section and axillary dissection was performed only in cases of positive SLNB. RESULTS: The median age of the patients was 48 (range, 22-81) years. The mean number of hot spots identified was 1.4 in the whole series, 1.3 in patients who received a single injection, and 1.7 in those who received a double injection (P < 0.001). The mean number of removed SLNs was 1.7 (median, 1; range, 1-7) with an identification rate of 100%. A total of 138 patients with negative SLNB (n = 134) or isolated tumor cells in the SLN (n = 4) did not receive completion axillary lymph node dissection (CALND). In these latter patients, a total of 27 events (19.5%) occurred with 3 patients (2.2%) developing axillary recurrences after a median follow-up of 5 years (range, 17-134 months). CONCLUSIONS: Axillary lymph node reappearance was infrequent among patients with multicentric breast cancer, having negative SLNB and no CALND. We recommend SLNB as the standard procedure for nodal staging in patients with multicentric breast cancer and a clinically negative axilla.

The Changing Face of Mastectomy (from Mutilation to Aid to Breast Reconstruction).

Breast cancer is the most common cancer in women. Primary treatment is surgery, with mastectomy as the main treatment for most of the twentieth century. However, over that time, the extent of the procedure varied, and less extensive mastectomies are employed today compared to those used in the past, as excessively mutilating procedures did not improve survival. Today, many women receive breast-conserving surgery, usually with radiotherapy to the residual breast, instead of mastectomy, as it has been shown to be as effective as mastectomy in early disease. The relatively new skin-sparing mastectomy, often with immediate breast reconstruction, improves aesthetic outcomes and is oncologically safe. Nipple-sparing mastectomy is newer and used increasingly, with better acceptance by patients, and again appears to be oncologically safe. Breast reconstruction is an important adjunct to mastectomy, as it has a positive psychological impact on the patient, contributing to improved quality of life.