Impact of primary kidney disease on the effects of empagliflozin in patients with chronic kidney disease: secondary analyses of the EMPA-KIDNEY trial

Background: The EMPA KIDNEY trial showed that empagliflozin reduced the risk of the primary composite outcome of kidney disease progression or cardiovascular death in patients with chronic kidney disease mainly through slowing progression. We aimed to assess how effects of empagliflozin might differ by primary kidney disease across its broad population. Methods: EMPA-KIDNEY, a randomised, controlled, phase 3 trial, was conducted at 241 centres in eight countries (Canada, China, Germany, Italy, Japan, Malaysia, the UK, and the USA). Patients were eligible if their estimated glomerular filtration rate (eGFR) was 20 to less than 45 mL/min per 1·73 m2, or 45 to less than 90 mL/min per 1·73 m2 with a urinary albumin-to-creatinine ratio (uACR) of 200 mg/g or higher at screening. They were randomly assigned (1:1) to 10 mg oral empagliflozin once daily or matching placebo. Effects on kidney disease progression (defined as a sustained ≥40% eGFR decline from randomisation, end-stage kidney disease, a sustained eGFR below 10 mL/min per 1·73 m2, or death from kidney failure) were assessed using prespecified Cox models, and eGFR slope analyses used shared parameter models. Subgroup comparisons were performed by including relevant interaction terms in models. EMPA-KIDNEY is registered with ClinicalTrials.gov, NCT03594110. Findings: Between May 15, 2019, and April 16, 2021, 6609 participants were randomly assigned and followed up for a median of 2·0 years (IQR 1·5–2·4). Prespecified subgroupings by primary kidney disease included 2057 (31·1%) participants with diabetic kidney disease, 1669 (25·3%) with glomerular disease, 1445 (21·9%) with hypertensive or renovascular disease, and 1438 (21·8%) with other or unknown causes. Kidney disease progression occurred in 384 (11·6%) of 3304 patients in the empagliflozin group and 504 (15·2%) of 3305 patients in the placebo group (hazard ratio 0·71 [95% CI 0·62–0·81]), with no evidence that the relative effect size varied significantly by primary kidney disease (pheterogeneity=0·62). The between-group difference in chronic eGFR slopes (ie, from 2 months to final follow-up) was 1·37 mL/min per 1·73 m2 per year (95% CI 1·16–1·59), representing a 50% (42–58) reduction in the rate of chronic eGFR decline. This relative effect of empagliflozin on chronic eGFR slope was similar in analyses by different primary kidney diseases, including in explorations by type of glomerular disease and diabetes (p values for heterogeneity all >0·1). Interpretation: In a broad range of patients with chronic kidney disease at risk of progression, including a wide range of non-diabetic causes of chronic kidney disease, empagliflozin reduced risk of kidney disease progression. Relative effect sizes were broadly similar irrespective of the cause of primary kidney disease, suggesting that SGLT2 inhibitors should be part of a standard of care to minimise risk of kidney failure in chronic kidney disease. Funding: Boehringer Ingelheim, Eli Lilly, and UK Medical Research Council

A simple approach for multicolor immunofluorescence staining in different Drosophila cell types

Multicolor immunostaining analysis is often a desirable tool in cell biology for most researchers. Nonetheless, this is not an easy task and often not affordable by many laboratories as it might require expensive instrumentation and sophisticated analysis software. Here, we describe a simple protocol for performing sequential immunostainings on two different Drosophila specimens. Our strategy relies on an efficient and reproducible method for removal primary antibodies and/or fluorophore-conjugated secondary antibodies that does not affect antigene integrity. We show that alternation of multiple rounds of antibody incubation and removal on the same slide, followed by registration of the same DAPI-stained image, provides a simple framework for the sequential detection of several antigens in the same cell. Given that the sample fixation procedures used for Drosophila tissues are compatible with most specimen processing protocols, we can envisage that the multicolor immunostaining strategy presented here can be also adapted to different samples including mammalian tissues and/or cells. J. Cell. Physiol. 229: 683-687, 2014. (c) 2013 Wiley Periodicals, Inc

Metrological assessment of TDR performance for moisture evaluation in granular materials

The monitoring of moisture levels and the investigation of the dielectric properties of materials are tasks that can be suitably performed through Time Domain Reflectometry (TDR) techniques. In particular, moisture monitoring of granular agro-foods is a major interest for several industrial applications. Despite the extensive literature regarding the use of TDR technology, the state of the art is lacking in a rigorous metrological characterization of the results achieved, especially for the agro-foods area. To fill this gap, in this paper the Authors carry out a comparative analysis between moisture content measurements obtained through two different TDR instruments on various granular materials. First, the major error contributions affecting the overall performance of the TDR-based method are discussed; in particular, the error contributions that are intrinsic in the method are discriminated from those related to the specific experimental conditions. Successively, systematic errors effects are minimized through a preliminary calibration procedure of the experimental set-up. Finally, results are compared and analyzed through a statistical approach, providing both an assessment of the repeatability of measurements and a rigorous metrological characterization. This ultimately leads to a performance characterization, particularly useful for practical industrial applications

TDR Moisture Estimation for Granular Materials: An Application in Agro-food Industrial Monitoring

Time-domain reflectometry (TDR) is a wellestablished technique for continuous monitoring of the moisture levels and dielectric properties of materials; typically, it is widely used in soil science applications. Despite the high flexibility, elevated accuracy, and the low cost of TDR instrumentation, the state of the art is actually rather lacking in specific applications for the quality monitoring of agro-industrial processes and agro-food products. Therefore, the possibility of real-time control of moisture level plays a crucial role in terms of optimization, quality preservation, and energy saving in several industrial applications. In this regard, the development of suitable models relating the\ud dielectric properties of granular materials to their moisture content is a still an open issue. To fill this gap, in this paper, the authors study the feasibility of adopting the TDR technique for such a purpose. The proposed goal is successfully reached through a comparative analysis among different calibration procedures and through experimental measurements on various granular materials. Finally, the adopted methodology is assessed through a rigorous metrological characterization, thus allowing the evaluation of the associated measurement uncertainty and repeatability

TDR Moisture Measurements in Granular Materials: from the Siliceous Sand test-case to the Applications for Agro-food Industrial Monitoring

Different approaches are commonly available for the continuous monitoring of moisture and dielectric properties of various materials. In particular Time Domain Reflectometry (TDR) is the standard method for measuring soil water content. Nevertheless, the state of the art is rather lacking in assessing specific calibration procedures allowing the accurate moisture monitoring of granular materials. To fill this gap, the adoption of TDR for such purposes is investigated. Starting from measurements on siliceous sand, an in-depth analysis is extended to various feedstuff materials (such as corn, corn flour and bran). Furthermore, the adopted methodology is fully assessed from a metrological point of view, thus confirming its suitability for the considered applications

Metrological assessment of TDR performance for moisture evaluation in granular materials

The monitoring of moisture levels and the investigation of the dielectric properties of materials are tasks that can be suitably performed through Time Domain Reflectometry (TDR) techniques. In particular, moisture monitoring of granular agro-foods is a major interest for several industrial applications. Despite the extensive literature regarding the use of TDR technology, the state of the art is lacking in a rigorous metrological characterization of the results achieved, especially for the agro-foods area. To fill this gap, in this paper the Authors carry out a comparative analysis between moisture content measurements obtained through two different TDR instruments on various granular materials. First, the major error contributions affecting the overall performance of the TDR-based method are discussed; in particular, the error contributions that are intrinsic in the method are discriminated from those related to the specific experimental conditions. Successively, systematic errors effects are minimized through a preliminary calibration procedure of the experimental set-up. Finally, results are compared and analyzed through a statistical approach, providing both an assessment of the repeatability of measurements and a rigorous metrological characterization. This ultimately leads to a performance characterization, particularly useful for practical industrial applications. © 2008 Elsevier Ltd. All rights reserved

Tachycardia and coronary blood flow: non-invasive estimation during Valsalva manoeuvre and exercise

Simultaneous changes in cycle length and coronary blood flow were studied during Valsalva manoeuvre and supine cycloergometer exercise test in 10 male patients (mean age 48+/-12 years) who had successfully undergone myocardial revascularization by surgical anastomosis of the left internal mammary artery on the left anterior descending coronary artery. Blood velocity curves in the left internal mammary artery were obtained by a non-invasive continuous-wave Doppler probe at rest, in the last phase of the expiratory effort of the Valsalva manoeuvre and at the maximum load attained during the exercise test. Mean arterial pressure by sphygmomanometer, and cardiac cycle length on the basis of Doppler recording were measured. Mean blood velocity, the length of the blood column entering the coronary bed at each cycle (cardiac cycle times mean velocity), an index of blood cell acceleration (the ratio of mean velocity to cardiac cycle), and an index of coronary resistance (the ratio of mean pressure to mean velocity), were calculated

A Non-Invasive Approach for Moisture Measurements through Patch Antennas

Different approaches and techniques are commonly available for the continuous monitoring of moisture levels and dielectric properties of materials; typically, related applications, are largely used in soil science area. Nevertheless, the state of the art is actually rather lacking in specific sensing methods that combine the low cost and the non-invasive approach of the involved instrumentation. To fill this gap, in this paper the authors study the feasibility of the adoption of the time-domain reflectometry analysis for such purposes, using a patch antenna as sensing element. This way, the extrapolation of the scattering parameters is suitably correlated to the water content levels of the materials under test. Furthermore, the adopted methodology is assessed through full-wave simulations, thus corroborating the experimental measurements