Role of zebrafish ClC-K/barttin channels in apical kidney chloride reabsorption.

KEY POINTS: We have characterized the zebrafish clc-k and barttin proteins, demonstrating that they form a protein complex mediating chloride flux in a similar manner to their mammalian counterparts. As in mammals, in zebrafish, clc-k and barttin are basically expressed in the kidney. Contrary to what is found in mammals, in zebrafish both proteins show an apical localization in the kidney. We have generated the first knockout in zebrafish of a CLC protein. Lack of clc-k in zebrafish resulted in embryonic lethality, possibly caused by a reduction in total chloride content. As a consequence, there is an up-regulation of other chloride channels and other regulatory mechanisms such as renin or the uro-guanylin receptor in the kidney. barttin is mislocalized in vivo when clc-k is not present, indicating that there is a mutual dependence of the protein expression and localization between barttin and clc-k proteins. ABSTRACT: ClC-K/barttin channels are very important for salt transport in the kidney. This function can be clearly seen since mutations in CLCNKB or BSND cause Bartter's syndrome types III and IV, respectively. Working with the freshwater teleost zebrafish, we characterized the genes homologous to the mammalian chloride channel ClC-K and its obligate subunit barttin and we obtained and studied clc-k knockout zebrafish. The zebrafish clc-k/barttin proteins are very similar to their mammalian counterparts, and both proteins are necessary to mediate chloride currents. Localization studies indicated that both proteins are exclusively expressed in the apical membranes of zebrafish kidney tubules. Knockout of clc-k resulted in embryonic lethality. These animals showed barttin mislocalization and a reduction in whole-body chloride concentration, as well as up-regulation of the expression of other chloride channels and renin, and an increase in the kidney expression of the uroguanylin receptor. Our results indicate that apical kidney chloride reabsorption through clc-k/barttin channels is crucial for chloride homeostasis in zebrafish as it is in humans. The zebrafish model could be used as a new in vivo system to study ClC-K function.

Inhibitory killer cell immunoglobulin-like receptor (iKIR) mismatches improve survival after T-cell-repleted haploidentical transplantation.

Alloreactivity triggered by interaction between killer cell immunoglobulin-like receptors (KIRs) and natural killer (NK) cells plays a role in the graft-versus-tumor effect after hematopoietic stem cell transplantation (SCT). Our aim in this study was to evaluate this role in the setting of T-cell-repleted haploidentical SCT with postinfusion high-dose cyclophosphamide (PT-Cy). We included 33 patients. Among patient-donor pairs with at least 1 inhibitory KIR (iKIR) gene mismatch, event-free survival (EFS) and cumulative incidence of relapse 1 year after transplant were significantly better (85% vs. 37% [P = 0.008] and 18% vs. 46% [P = 0.041], respectively). A subanalysis in 12 patients with Hodgkin's lymphoma (HL) showed an improvement in EFS 1 year after transplant in those patients with KIR ligand mismatch (100% vs. 25%, P = 0.012), although overall survival (OS) was not affected (85% vs. 80%, P = 0.2). Eight of 12 patient-donors pairs presented iKIR mismatches. Of note, this outcome was better in the small subgroup, both for EFS (100% vs. 25%, P = 0.012) and for OS (100% vs. 37%, P = 0.004). Our data suggest that in the setting of T-cell-repleted haploidentical SCT with PT-Cy, iKIR mismatch is associated with improved survival, with particularly good results for both iKIR and KIR ligand mismatches in patients with HL.

Ibrutinib followed by ofatumumab consolidation in previously untreated patients with chronic lymphocytic leukemia (CLL): GELLC-7 trial from the Spanish group of CLL (GELLC).

Background: BTK inhibitors have been concurrently administered with anti-CD20 monoclonal antibodies (mAbs) in chronic lymphocytic leukemia (CLL). However, the optimal regimen for combining these two drugs remains pending. Methods: This multi-center phase 2 study aimed to analyze whether consolidation with ofatumumab improved the response in patients with CLL receiving front-line treatment with ibrutinib. Patients received 12 cycles of ibrutinib monotherapy. Those who achieved CR after this induction were maintained on ibrutinib. Conversely, those who did not attain CR continued with ibrutinib in addition to a consolidation, which involved 7 doses of ofatumumab. The primary objective was the complete response (CR) rate at cycle 20. This study is registered within the EU Clinical Trials Register (EudraCT 2016-004937-26). Findings: Between September 8, 2017, and May 21, 2018, 84 patients (median age, 69 years) were included. After completion of 12 cycles of ibrutinib (n = 80), 4 patients (5%) were in CR, 67 (84%) in partial response (PR), and 6 patients (7%) had a PR with lymphocytosis (PRL). After consolidation with ofatumumab, 20 patients improved the response from PR to CR and 6 patients with PRL obtained a PR. Seventy-one patients (85%) completed 20 cycles of treatment, with a CR rate of 24/71 (34%). According to the intention-to-treat analysis at cycle 20, the ORR was 69/84 (82.2%), with a CRR of 24/84 (28.6%). Progression-free survival and overall survival at 48-months were 89.9% (CI: 82.4-95.5) and 92.2% (CI: 85.3-97.1), respectively. Interpretation: These findings underscore the potential for a consolidation strategy in CLL, wherein the addition of a mAb in patients with low tumor burden might enhance the quality of the response. Funding: The study was funded by Janssen that also supplied ibrutinib, whereas ofatumumab was supplied by Novartis.

Diabetic foot self-care and concordance of 3diabetic foot risk stratification systems in a basic health area of Gran Canaria. / Nivel de autocuidados en pie diabético y concordancia de 3sistemas de estratificación de riesgo en una zona básica de salud de Gran Canaria.

OBJECTIVE: To assess the level of self-care in the population with diabetes and determine the risk of diabetic foot lesions through the use of 3stratification systems as well as to establish the degree of concordance between these systems. METHOD: Observational, cross-sectional and descriptive study carried out in the Basic Health Area of Santa Brígida (Gran Canaria-Canary Islands-Spain) in people diagnosed with diabetes (DM Type 1/DM Type 2) (n=182). Interview, physical examination, review of clinical history and completion of the Diabetic Foot Self-Care questionnaire of the University of Malaga were carried out. The risk stratification was then calculated using 3systems (System of the National Institute for Health Care Excellence, Classification of the International Working Group on the Diabetic Foot and High-Risk Diabetic Foot-60-Second Tool©-2012). The Kappa index was calculated to study the concordance between systems, the relative risk of negative screening of one method against another was estimated and the exact Fisher test to establish whether there were differences. RESULTS: 30.2% of diabetics had a low level of self-care, 45.1% a medium level and 24.7% a high level. The risk levels calculated were: National Institute for Health Care Excellence Classification (Negative Risk 71.4%-Positive Risk 28.6%), International Working Group on the Diabetic Foot Classification (Negative Risk 67.0%-Positive Risk 33.0%) and High-Risk Diabetic Foot-60-Second Tool© (Negative Risk 62.6%-Positive Risk 37.4%). CONCLUSIONS: All 3systems have good concordance with each other. The High-Risk Diabetic Foot-60-Second Tool© only distinguishes 2levels of risk but detects a higher percentage of people at risk. The Diabetic Foot Self-Care questionnaire of the University of Malaga may be useful in the context of Primary Care to assess the level of self-care of people with diabetes.

Nivel de autocuidados en pie diabético y concordancia de 3 sistemas de estratificación de riesgo en una zona básica de salud de Gran Canaria / Diabetic foot self-care and concordance of 3diabetic foot risk stratification systems in a basic health area of Gran Canaria

Objetivo: Evaluar el nivel de autocuidados en la población con diabetes y determinar el riesgo de padecer lesiones de pie diabético mediante el uso de 3 sistemas de estratificación, así como establecer el grado de concordancia entre estos sistemas. Método: Estudio observacional, transversal y descriptivo realizado en la Zona básica de salud de Santa Brígida (Gran Canaria, Islas Canarias, España) en personas diagnosticadas de diabetes (DM tipo 1/DM tipo 2) (n = 182). Se realizaron entrevista, exploración física, revisión de la historia clínica y cumplimentación del cuestionario Diabetic Foot Self-Care questionnaire of the University of Malaga. Tras ello se calculó la estratificación del riesgo con 3 sistemas (sistema del National Institute for Health Care Excellence, clasificación del International Working Group on the Diabetic Foot y High-Risk Diabetic Foot-60-Second Tool© 2012). Se calculó el índice kappa para estudiar la concordancia entre sistemas, se estimaron el riesgo relativo de screening negativo de un método frente a otro y el test exacto de Fisher para establecer si existían diferencias. Resultados: Un 30,2% de los diabéticos tenían un nivel bajo de autocuidados, un 45,1% un nivel medio y un 24,7% nivel alto. Los niveles de riesgo calculados fueron: fueron clasificación National Institute for Health Care Excellence (riesgo negativo 71,4%; riesgo positivo 28,6%), clasificación del International Working Group on the Diabetic Foot (riesgo negativo 67,0%; riesgo positivo 33,0%) y High-Risk Diabetic Foot-60-Second Tool© (riesgo negativo 62,6%; riesgo positivo 37,4%). Conclusiones: Los 3 sistemas poseen una buena concordancia entre sí. El High-Risk Diabetic Foot-60-Second Tool© solo distingue 2 niveles de riesgo pero detecta mayor porcentaje de personas en situación de riesgo. El cuestionario Diabetic Foot Self-Care questionnaire of the University of Malaga puede ser útil en el contexto de Atención Primaria para evaluar el nivel de autocuidados de las personas con diabetes

Objective: To assess the level of self-care in the population with diabetes and determine the risk of diabetic foot lesions through the use of 3 stratification systems as well as to establish the degree of concordance between these systems. Method: Observational, cross-sectional and descriptive study carried out in the Basic Health Area of Santa Brígida (Gran Canaria-Canary Islands-Spain) in people diagnosed with diabetes (DM Type 1/DM Type 2) (n = 182). Interview, physical examination, review of clinical history and completion of the Diabetic Foot Self-Care questionnaire of the University of Malaga were carried out. The risk stratification was then calculated using 3systems (System of the National Institute for Health Care Excellence, Classification of the International Working Group on the Diabetic Foot and High-Risk Diabetic Foot-60-Second Tool©-2012). The Kappa index was calculated to study the concordance between systems, the relative risk of negative screening of one method against another was estimated and the exact Fisher test to establish whether there were differences. Results: 30.2% of diabetics had a low level of self-care, 45.1% a medium level and 24.7% a high level. The risk levels calculated were: National Institute for Health Care Excellence Classification (Negative Risk 71.4%-Positive Risk 28.6%), International Working Group on the Diabetic Foot Classification (Negative Risk 67.0%-Positive Risk 33.0%) and High-Risk Diabetic Foot-60-Second Tool© (Negative Risk 62.6%-Positive Risk 37.4%). Conclusions: All 3 systems have good concordance with each other. The High-Risk Diabetic Foot-60-Second Tool© only distinguishes 2 levels of risk but detects a higher percentage of people at risk. The Diabetic Foot Self-Care questionnaire of the University of Malaga may be useful in the context of Primary Care to assess the level of self-care of people with diabetes