Identification of hereditary breast and ovarian cancer germline variants in Granada (Spain): NGS perspective.

The aim of this study was to assess the prevalence of germline variants in cancer-predisposing genes by either targeted (BRCA1/2) or multigene NGS panel in a high-risk Hereditary Breast and Ovarian Cancer (HBOC) cohort. Samples from 824 Caucasian probands were retrospectively collected and the impact of genetic diagnosis and genetic variants epidemiology in this cohort was evaluated. Performance of risk-reducing prophylactic measures, such as prophylactic mastectomy and/or prophylactic oophorectomy, was assessed through clinical follow-up of patients with a positive genetic result. Pathogenic variants predisposing to HBOC were identified in 11.9% (98/824) individuals at BRCA2 (47/98), BRCA1 (24/98), PALB2 (8/51), ATM (7/51), CHEK2 (6/51) MSH6, (2/51), RAD51C (2/51) and TP53 (2/386). Of them, 11 novel pathogenic variants and 12 VUS were identified, characterized, and submitted to ClinVar. Regarding clinical impact, the risk of developing basal or Her2 breast cancer was increased 15.7 times or 37.5 times for BRCA1 and MSH6 pathogenic variants respectively. On the contrary, the risk of developing basal or luminal A breast cancer was reduced to 81% or 77% for BRCA2 and BRCA1 pathogenic variants, respectively. Finally, 53.2% of individuals testing positive for class IV/V variants underwent prophylactic surgery (mastectomy, oophorectomy or both) being significantly younger at the cancer diagnosis than those undertaking prophylactic measures (p = 0.008). Of them, 8 carried a pathogenic/likely pathogenic variant in other genes different from BRCA1 and BRCA2, and the remaining (46.7%) decided to continue with clinical follow-up. No differences in pathogenicity or risk of developing cancer were found for BRCA1/2 between targeted and multigene sequencing strategies; however, NGS was able to resolve a greater proportion of high-risk patients.

Deep Phenotypic Characterisation of CTCs by Combination of Microfluidic Isolation (IsoFlux) and Imaging Flow Cytometry (ImageStream).

The isolation of circulating tumour cells (CTCs) in colorectal cancer (CRC) mostly relies on the expression of epithelial markers such as EpCAM, and phenotypic characterisation is usually performed under fluorescence microscopy with only one or two additional markers. This limits the ability to detect different CTC subpopulations based on multiple markers. The aim of this work was to develop a novel protocol combining two platforms (IsoFluxTM and ImageStream®X) to improve CTC evaluation. Cancer cell lines and peripheral blood from healthy donors were used to evaluate the efficiency of each platform independently and in combination. Peripheral blood was extracted from 16 early CRC patients (before loco-regional surgery) to demonstrate the suitability of the protocol for CTC assessment. Additionally, peripheral blood was extracted from nine patients one month after surgery to validate the utility of our protocol for identifying CTC subpopulation changes over time. Results: Our protocol had a mean recovery efficiency of 69.5% and a limit of detection of at least four cells per millilitre. We developed an analysis method to reduce noise from magnetic beads used for CTC isolation. CTCs were isolated from CRC patients with a median of 37 CTCs (IQ 13.0-85.5) at baseline. CTCs from CRC patients were significantly (p < 0.0001) larger than cytokeratin (CK)-negative cells, and patients were stratified into two groups based on BRAFV600E and PD-L1 expression on CK-positive cells. The changes observed over time included not only the number of CTCs but also their distribution into four different subpopulations defined according to BRAFV600E and PD-L1 positivity. We developed a novel protocol for semi-automatic CTC isolation and phenotypic characterisation by combining two platforms. Assessment of CTCs from early CRC patients using our protocol allowed the identification of two clusters of patients with changing phenotypes over time.

The Polemic Diagnostic Role of TP53 Mutations in Liquid Biopsies from Breast, Colon and Lung Cancers.

Being minimally invasive and thus allowing repeated measures over time, liquid biopsies are taking over traditional solid biopsies in certain circumstances such as those for unreachable tumors, very early stages or treatment monitoring. However, regarding TP53 mutation status analysis, liquid biopsies have not yet substituted tissue samples, mainly due to the lack of concordance between the two types of biopsies. This needs to be examined in a study-dependent manner, taking into account the particular type of liquid biopsy analyzed, that is, circulating tumor cells (CTCs) or cell-free DNA (cfDNA), its involvement in the tumor biology and evolution and, finally, the technology used to analyze each biopsy type. Here, we review the main studies analyzing TP53 mutations in either CTCs or cfDNA in the three more prevalent solid tumors: breast, colon and lung cancers. We evaluate the correlation for mutation status between liquid biopsies and tumor tissue, suggesting possible sources of discrepancies, as well as evaluating the clinical utility of using liquid biopsies for the analysis of TP53 mutation status and the future actions that need to be undertaken to make liquid biopsy analysis a reality for the evaluation of TP53 mutations.

Circulating tumor cells criteria (CyCAR) versus standard RECIST criteria for treatment response assessment in metastatic colorectal cancer patients.

BACKGROUND: The use of circulating tumor cells (CTCs) as indicators of treatment response in metastatic colorectal cancer (mCRC) needs to be clarified. The objective of this study is to compare the Response Evaluation Criteria in Solid Tumors (RECIST) with the Cytologic Criteria Assessing Response (CyCAR), based on the presence and phenotypic characterization of CTCs, as indicators of FOLFOX-bevacizumab treatment response. METHODS: 77 mCRC blood samples from FOLFOX-bevacizumab treated patients were analyzed to isolate CTCs before and after (12 and 24 weeks) treatment, using an immunomagnetic separation method. VEGFR expression was identified by double immunostaining. RESULTS: We observed a decrease of CTCs (42.8 vs. 18.2%) and VEGFR positivity (69.7% vs. 41.7%) after treatment. According to RECIST, 6.45% of the patients did not show any clinical benefit, whereas 93.55% patients showed a favorable response at 12 weeks. According to CyCAR, 29% had a non-favorable response and 71% patients did not. No significant differences were found between the response assessment by RECIST and CyCAR at 12 or 24 weeks. However, in the multivariate analysis, RECIST at 12 weeks and CyCAR at 24 weeks were independent prognostic factors for OS (HR: 0.1, 95% CI 0.02-0.58 and HR: 0.35, 95% CI 0.12-0.99 respectively). CONCLUSIONS: CyCAR results were comparable to RECIST in evaluating the response in mCRC and can be used as an alternative when the limitation of RECIST requires additional response analysis techniques.

CD133 expression in circulating tumor cells from breast cancer patients: potential role in resistance to chemotherapy.

CD133 has been associated with cell properties such as self renewal, migration and vasculogenic mimicry, potentially involved in generation of circulating tumor cells (CTCs). We characterized CD133 expression in CTCs of 98 nometastatic breast cancer (BC) patients. CTCs were isolated by immunomagnetic techniques using magnetic beads labeled with a multicytokeratin(CK)-specific antibody (CK3-11D5) and CTCs and CD133 detection through immunocytochemical methods. CK(+) /CD133(+) CTCs were identified in 65% of patients at baseline and 47.8% after systemic therapy (p = 0.53). Correlation of CD133 status in CTCs with classical clinicopathological characteristics and response to therapy was performed. Her2 not amplified and low Ki-67 index were positively correlated with presence of CK(+) /CD133(+) CTCs. Before any treatment, CK(+) /CD133(+) CTCs were more frequently isolated in patients with luminal BC subtype. No statistically significant differences were found between proportion of CK(+) /CD133(+) CTCs and BC subtypes after systemic therapy, implying a relative enrichment of CK(+) /CD133(+) CTCs in triple negative and HER2-amplified tumors. While CK(+) /CTCs decreases after chemotherapy when analyzing the whole population, CK(+) /CD133(+) CTCs were enriched in post-treatment samples in nonluminal BC subtypes. These findings suggest the potential role of CD133 as a promising marker of chemoresistance in nonluminal BC patients. Further prospective studies and extensive preclinical modeling will be needed to confirm whether CD133 is a marker of resistance to chemotherapy, and its role as a target for novel anticancer therapies targeting cancer stem cells and tumor vasculature.

Satisfacción de los usuarios con el servicio de teleoftalmología con cámara no midriática para el cribado de la retinopatía diabética / User satisfaction with teleophthalmology with nonmydriatic camera for diabetic retinopathy screening

ObjetivoConocer la satisfacción de los pacientes diabéticos respecto al servicio de retinografía.MétodosEncuesta telefónica a 64 usuarios entre julio de 2006 y marzo de 2007. El 57,8% eran varones. La edad media fue de 65,2 años. El 54,7% procedían de un equipo de atención primaria (EAP) urbano. Las variables fueron: sexo, edad, EAP, retinografía/tonometría (normal/patológica), accesibilidad, puntualidad, limpieza, tiempo dedicado, explicaciones, buenas manos, amabilidad en escala mala/regular/buena/muy buena/perfecta, y satisfacción telefónica/global en escala 0–10.ResultadosSe valoró (>80%) la accesibilidad, la puntualidad, la limpieza, el tiempo dedicado, las buenas manos y la amabilidad. La media de satisfacción global fue de 8,38 (intervalo de confianza del 95% [IC95%]: 8,03–8,72) y la telefónica fue de 7,88 (IC95%: 7,4–8,36). Se asociaron (p<0,05) con una satisfacción global (⩾8) el tiempo de visita, las explicaciones comprensibles y la llamada telefónica que informa del resultado. La regresión logística muestra (p<0,05) que en la satisfacción influye más recibir el resultado de las exploraciones por teléfono.ConclusionesEl servicio de retinografía está bien valorado. Los resultados telefónicos son los que más influyen en la elevada satisfacción. Se impulsarán las nuevas tecnologías (SMS, e-mail)(AU)

ObjectiveTo determine satisfaction with the retinography service among patients with diabetes.MethodsWe performed a telephone survey of 64 users from July 2006 to March 2007. The mean age was 65.2 years, 57.8% were men, and 54.7% were from urban primary care centers. The variables analyzed were sex, age, primary care team, retinography/tonometry (normal/pathologic), accessibility, punctuality, hygiene, consultation length, explanations, good hands, kindness on a scale rated bad/average/good/very good/perfect, satisfaction with the telephone call informing users of the results of the examination, and overall satisfaction, both rated on a scale from 0 to 10.ResultsAccessibility, punctuality, hygiene, consultation length, good hands and kindness received scores of >80%. The mean overall satisfaction score was 8.38% (95% confidence interval [95%CI]: 8.03–8.72), while satisfaction with the telephone call was 7.88% (95%CI: 7.4–8.36). The variables associated (p<0.05) with overall satisfaction of ⩾8 were consultation length, receiving comprehensible explanations, and the telephone call informing patients of the results of the examination. Logistic regression showed (p<0.05) that the variable with the greatest influence on satisfaction was the telephone call.Conclusionsthe retinography service was favorably evaluated. The variable with the greatest influence on high satisfaction was communicating the results by telephone. The service will promote new technologies (SMS, e-mail)(AU)

Fístula arteriovenosa renal congénita / Arteriovenous renal congenital fístula

Presentamos el caso de una mujer joven que acudió a urgencias con un cuadro de hematuria macroscópica anemizante. A su vez confirmamos que ante la sospecha de encontrarnos ante una fístula arteriovenosa, la arteriografía es la prueba diagnóstica de elección y la embolización selectiva, la mejor opción terapéutica (AU)

We present the case of a young woman who went to the Emergency Departent with macroscopic and anaemic haematuria. We also confirm that, when faced with the possibility of finding an arteriovenous fistula, arteriography is the best choice for diagnosis, and the best option for treatment is selective embolization (AU)

[User satisfaction with teleophthalmology with nonmydriatic camera for diabetic retinopathy screening]. / Satisfacción de los usuarios con el servicio de teleoftalmología con cámara no midriática para el cribado de la retinopatía diabética.

OBJECTIVE: To determine satisfaction with the retinography service among patients with diabetes. METHODS: We performed a telephone survey of 64 users from July 2006 to March 2007. The mean age was 65.2 years, 57.8% were men, and 54.7% were from urban primary care centers. The variables analyzed were sex, age, primary care team, retinography/tonometry (normal/pathologic), accessibility, punctuality, hygiene, consultation length, explanations, good hands, kindness on a scale rated bad/average/good/very good/perfect, satisfaction with the telephone call informing users of the results of the examination, and overall satisfaction, both rated on a scale from 0 to 10. RESULTS: Accessibility, punctuality, hygiene, consultation length, good hands and kindness received scores of >80%. The mean overall satisfaction score was 8.38% (95% confidence interval [95%CI]: 8.03-8.72), while satisfaction with the telephone call was 7.88% (95%CI: 7.4-8.36). The variables associated (p<0.05) with overall satisfaction of 8 were consultation length, receiving comprehensible explanations, and the telephone call informing patients of the results of the examination. Logistic regression showed (p<0.05) that the variable with the greatest influence on satisfaction was the telephone call. CONCLUSIONS: the retinography service was favorably evaluated. The variable with the greatest influence on high satisfaction was communicating the results by telephone. The service will promote new technologies (SMS, e-mail).