Directorate-General Joint Research Centre

BACKGROUND: These recommendations have been developed to improve the care of intensive care unit (ICU) patients during the dying process. The recommendations build on those published in 2003 and highlight recent developments in the field from a U.S. perspective. They do not use an evidence grading system because most of the recommendations are based on ethical and legal principles that are not derived from empirically based evidence. PRINCIPAL FINDINGS: Family-centered care, which emphasizes the importance of the social structure within which patients are embedded, has emerged as a comprehensive ideal for managing end-of-life care in the ICU. ICU clinicians should be competent in all aspects of this care, including the practical and ethical aspects of withdrawing different modalities of life-sustaining treatment and the use of sedatives, analgesics, and nonpharmacologic approaches to easing the suffering of the dying process. Several key ethical concepts play a foundational role in guiding end-of-life care, including the distinctions between withholding and withdrawing treatments, between actions of killing and allowing to die, and between consequences that are intended vs. those that are merely foreseen (the doctrine of double effect). Improved communication with the family has been shown to improve patient care and family outcomes. Other knowledge unique to end-of-life care includes principles for notifying families of a patient's death and compassionate approaches to discussing options for organ donation. End-of-life care continues even after the death of the patient, and ICUs should consider developing comprehensive bereavement programs to support both families and the needs of the clinical staff. Finally, a comprehensive agenda for improving end-of-life care in the ICU has been developed to guide research, quality improvement efforts, and educational curricula. CONCLUSIONS: End-of-life care is emerging as a comprehensive area of expertise in the ICU and demands the same high level of knowledge and competence as all other areas of ICU practice.

SUMMARY: When Saccharomyces cerevisiae is growing exponentially on glucose or fructose as carbon plus energy source, and in the presence of air, the glucose degradation proceeds mainly via aerobic fermentation. When the yeast is growing on mannose or galactose, degradation proceeds simultaneously via respiration and fermentation. This situation results from a repression of the of the respiratory enzymes synthesis by high fermentation rates. This regulatory system, called the “Crabtree effect”, consists actually of a repression of an energy source (respiration) by another energy source (fermentation). Various yeast strains were tested; the regulatory system was present in about 50% of them.

Whereas the dramatic environmental impact of plastic waste rightfully receives considerable attention by scientists, policy makers and public in general, the human health impact of micro- and nanoplastics contamination of our food and beverages remains largely unknown. Indeed, most studies aim at understanding the environmental impact rather than the human health impact of a possible exposure to micro- and nanoplastics. In addition, these papers generally lack a methodological, standardised approach. Furthermore, some studies focus on the damage to and contamination level of animal species collected from the wild environment, and others investigate the rate and biology of microplastic uptake of animals fed with microplastics in laboratory. This review aims at understanding human exposure. Since there is, with few exceptions, no evidence available on the presence of micro- and nanoplastics in a normal diet, this study takes an indirect approach and analyses peer-reviewed publications since 2010 that document the presence of micro- and nanoplastics in those animals (more than 200 species) and food products that are part of the human food chain and that may thus contribute directly or indirectly to the uptake of micro- and nanoplastics via the human diet. It also addresses the question of the definitions, the methodologies and the quality criteria applied to obtain the reported results. This review suggests that, beyond a few estimations and comparisons, precise data to assess the exact exposure of humans to micro- and nanoplastics through their diet cannot be produced until standardised methods and definitions are available.

This article develops an information criterion for determining the number q of common shocks in the general dynamic factor model developed by Forni et al., as opposed to the restricted dynamic model considered by Bai and Ng and by Amengual and Watson. Our criterion is based on the fact that this number q is also the number of diverging eigenvalues of the spectral density matrix of the observations as the number n of series goes to infinity. We provide sufficient conditions for consistency of the criterion for large n and T (where T is the series length). We show how the method can be implemented and provide simulations and empirics illustrating its very good finite-sample performance. Application to real data adds a new empirical facet to an ongoing debate on the number of factors driving the U.S. economy.

College students (N = 3,435) in 26 cultures reported their perceptions of age-related changes in physical, cognitive, and socioemotional areas of functioning and rated societal views of aging within their culture. There was widespread cross-cultural consensus regarding the expected direction of aging trajectories with (a) perceived declines in societal views of aging, physical attractiveness, the ability to perform everyday tasks, and new learning; (b) perceived increases in wisdom, knowledge, and received respect; and (c) perceived stability in family authority and life satisfaction. Cross-cultural variations in aging perceptions were associated with culture-level indicators of population aging, education levels, values, and national character stereotypes. These associations were stronger for societal views on aging and perceptions of socioemotional changes than for perceptions of physical and cognitive changes. A consideration of culture-level variables also suggested that previously reported differences in aging perceptions between Asian and Western countries may be related to differences in population structure.

Assessment of ecological status for the European Water Framework Directive (WFD) is based on “Biological Quality Elements” (BQEs), namely phytoplankton, benthic flora, benthic invertebrates and fish. Morphological identification of these organisms is a time-consuming and expensive procedure. Here, we assess the options for complementing and, perhaps, replacing morphological identification with procedures using eDNA, metabarcoding or similar approaches. We rate the applicability of DNA-based identification for the individual BQEs and water categories (rivers, lakes, transitional and coastal waters) against eleven criteria, summarised under the headlines representativeness (for example suitability of current sampling methods for DNA-based identification, errors from DNA-based species detection), sensitivity (for example capability to detect sensitive taxa, unassigned reads), precision of DNA-based identification (knowledge about uncertainty), comparability with conventional approaches (for example sensitivity of metrics to differences in DNA-based identification), cost effectiveness and environmental impact. Overall, suitability of DNA-based identification is particularly high for fish, as eDNA is a well-suited sampling approach which can replace expensive and potentially harmful methods such as gill-netting, trawling or electrofishing. Furthermore, there are attempts to replace absolute by relative abundance in metric calculations. For invertebrates and phytobenthos, the main challenges include the modification of indices and completing barcode libraries. For phytoplankton, the barcode libraries are even more problematic, due to the high taxonomic diversity in plankton samples. If current assessment concepts are kept, DNA-based identification is least appropriate for macrophytes (rivers, lakes) and angiosperms/macroalgae (transitional and coastal waters), which are surveyed rather than sampled. We discuss general implications of implementing DNA-based identification into standard ecological assessment, in particular considering any adaptations to the WFD that may be required to facilitate the transition to molecular data.

We examine the role of rainfall trends in poor growth performance of sub-Saharan African nations relative to other developing countries, using a new cross-country panel climatic data set in an empirical economic growth framework. Our results show that rainfall has been a significant determinant of poor economic growth for African nations but not for other countries. Depending on the benchmark measure of potential rainfall, we estimate that the direct impact under the scenario of no decline in rainfall would have resulted in a reduction of between around 15% and 40% of today's gap in African GDP per capita relative to the rest of the developing world.

Membranous nephropathy is characterized by deposition of immune complexes along the glomerular basement membrane. PLA2R and THSD7A are target antigens in 70% and 1-5% of primary membranous nephropathy cases, respectively. In the remaining cases, the target antigen is unknown. Here, laser microdissection of glomeruli followed by mass spectrometry was used to identify novel antigen(s) in PLA2R-negative membranous nephropathy. An initial pilot mass spectrometry study in 35 cases of PLA2R-negative membranous nephropathy showed high spectral counts for neural tissue encoding protein with EGF-like repeats, NELL-1, in six cases. Mass spectrometry failed to detect NELL-1 in 23 PLA2R-associated membranous nephropathy and 88 controls. NELL-1 was localized by immunohistochemistry, which showed bright granular glomerular basement membrane staining for NELL-1 in all six cases. Next, an additional 23 NELL-1 positive cases of membranous nephropathy were identified by immunohistochemistry in a discovery cohort of 91 PLA2R-negative membranous nephropathy cases, 14 were confirmed by mass spectrometry. Thus, 29 of 126 PLA2R-negative cases were positive for NELL-1. PLA2R-associated membranous nephropathy and controls stained negative for NELL-1. We then identified five NELL-1 positive cases of membranous nephropathy out of 84 PLA2R and THSD7A-negative cases in two validation cohorts from France and Belgium. By confocal microscopy, both IgG and NELL-1 co-localized to the glomerular basement membrane. Western blot analysis showed reactivity to NELL-1 in five available sera, but no reactivity in control sera. Clinical and biopsy findings of NELL-1 positive membranous nephropathy showed features of primary membranous nephropathy. Thus, a subset of membranous nephropathy is associated with accumulation and co-localization of NELL-1 and IgG along the glomerular basement membrane, and with anti-NELL-1 antibodies in the serum. Hence, NELL-1 defines a distinct type of primary membranous nephropathy. Membranous nephropathy is characterized by deposition of immune complexes along the glomerular basement membrane. PLA2R and THSD7A are target antigens in 70% and 1-5% of primary membranous nephropathy cases, respectively. In the remaining cases, the target antigen is unknown. Here, laser microdissection of glomeruli followed by mass spectrometry was used to identify novel antigen(s) in PLA2R-negative membranous nephropathy. An initial pilot mass spectrometry study in 35 cases of PLA2R-negative membranous nephropathy showed high spectral counts for neural tissue encoding protein with EGF-like repeats, NELL-1, in six cases. Mass spectrometry failed to detect NELL-1 in 23 PLA2R-associated membranous nephropathy and 88 controls. NELL-1 was localized by immunohistochemistry, which showed bright granular glomerular basement membrane staining for NELL-1 in all six cases. Next, an additional 23 NELL-1 positive cases of membranous nephropathy were identified by immunohistochemistry in a discovery cohort of 91 PLA2R-negative membranous nephropathy cases, 14 were confirmed by mass spectrometry. Thus, 29 of 126 PLA2R-negative cases were positive for NELL-1. PLA2R-associated membranous nephropathy and controls stained negative for NELL-1. We then identified five NELL-1 positive cases of membranous nephropathy out of 84 PLA2R and THSD7A-negative cases in two validation cohorts from France and Belgium. By confocal microscopy, both IgG and NELL-1 co-localized to the glomerular basement membrane. Western blot analysis showed reactivity to NELL-1 in five available sera, but no reactivity in control sera. Clinical and biopsy findings of NELL-1 positive membranous nephropathy showed features of primary membranous nephropathy. Thus, a subset of membranous nephropathy is associated with accumulation and co-localization of NELL-1 and IgG along the glomerular basement membrane, and with anti-NELL-1 antibodies in the serum. Hence, NELL-1 defines a distinct type of primary membranous nephropathy. see commentary on page 29 see commentary on page 29 Membranous nephropathy (MN) results from antibodies targeting an antigen in the glomerular basement membrane (GBM).1Beck Jr., L.H. Salant D.J. Membranous nephropathy: from models to man.J Clin Invest. 2014; 124: 2307-2314Crossref PubMed Scopus (109) Google Scholar, 2Ronco P. Debiec H. Pathophysiological advances in membranous nephropathy: time for a shift in patient's care.Lancet. 2015; 385: 1983-1992Abstract Full Text Full Text PDF PubMed Scopus (194) Google Scholar, 3Couser W.G. Primary membranous nephropathy.Clin J Am Soc Nephrol. 2017; 12: 983-997Crossref PubMed Scopus (245) Google Scholar The target antigen has been identified as M-type phospholipase A2 receptor (PLA2R) and thrombospondin type-1 domain-containing 7A (THSD7A) in approximately 70% and 1% to 5% of primary MN, respectively.4Beck Jr., L.H. Bonegio R.G. Lambeau G. et al.M-type phospholipase A2 receptor as target antigen in idiopathic membranous nephropathy.N Engl J Med. 2009; 361: 11-21Crossref PubMed Scopus (1419) Google Scholar, 5Tomas N.M. Hoxha E. Reinicke A.T. et al.Autoantibodies against thrombospondin type 1 domain-containing 7A induce membranous nephropathy.J Clin Invest. 2016; 126: 2519-2532Crossref PubMed Scopus (127) Google Scholar, 6Tomas N.M. Beck L.H. Meyer-Schwesinger C. et al.Thrombospondin type-1 domain-containing 7A in idiopathic membranous nephropathy.N Engl J Med. 2014; 371: 2277-2287Crossref PubMed Scopus (496) Google Scholar The target antigen(s) in the remaining PLA2R- and THSD7A-negative primary MN has remained elusive. The aim of this study was to identify an antigen(s) in the remaining primary MN. We used the novel methodology of laser microdissection and mass spectrometry to identify the major proteins in MN followed by immunostaining to localize and characterize the unique proteins. In the first step, we established that PLA2R antigen can be detected in PLA2R-positive MN. In the second step, we determined whether we could detect unique protein(s) with similar expression to PLA2R in a subset of PLA2R-negative MN. We initially selected 35 cases (pilot cohort) of PLA2R-negative MN on kidney biopsy for analysis by tandem mass spectrometry (MS/MS), and detected the unique protein, NELL-1, a neural tissue encoding protein with epidermal growth factor (EGF)-like repeats in 6 cases. We then analyzed the 35 cases of the pilot MS/MS and 91 additional PLA2R-negative MN cases by immunohistochemistry (IHC) for NELL-1 staining (discovery cohort). IHC confirmed the 6 positive NELL-1 cases of the pilot cohort and detected an additional 23 cases of NELL-1, bringing the total of NELL-1–positive cases to 29 (Figure 1). We performed MS/MS in 14 available samples of the 23 additional IHC NELL-1–positive cases from the discovery cohort to confirm the presence of NELL-1. Glomeruli were dissected (Figure 2a) and MS/MS studies from 35 PLA2R-negative MN cases (pilot cohort) detected the unique protein NELL-1 in 6 cases of the pilot cohort (Figure 2b). The average total spectral count for NELL-1 was 63.1 (SD ± 21.6) per case and is comparable to total spectral counts of PLA2R (86.1, SD ± 27.5) and Exostosin-1 (EXT1)/Exostosin-2 (EXT2) (EXT1: 65.3, SD ± 34.6; EXT2: 83.4, SD ± 38.4) in PLA2R-associated and EXT1/EXT2-associated MN, respectively.7Sethi S. Madden B.J. Debiec H. et al.Exostosin 1/exostosin 2–associated membranous nephropathy.J Am Soc Nephrol. 2019; 30: 1123-1136Crossref PubMed Scopus (107) Google Scholar All controls including PLA2R-associated MN cases were negative for NELL-1. MS/MS showed baseline spectral counts of PLA2R (average: 9.6, SD ± 8.6) in NELL-1–associated MN. The spectral counts of NELL-1 in the 6 cases, along with representative sequence coverage map of NELL-1 from 1 case are shown in Figure 2b and c. We subsequently performed MS/MS in 14 of 23 cases of the discovery cohort cases that were positive for NELL-1 by IHC. All cases showed similar high spectral counts of NELL-1 (Figure 2d). An example of MS/MS spectra match to a sequence from NELL-1 is shown in Supplementary Figure S1. All 4 classes of Igs were detected in NELL-1–associated MN: IgG1 was the most abundant Ig (average: 63.6, SD ± 13.1), followed by IgG3 (average: 53.2, SD ± 19.6), IgG2 (average: 50.6, SD ± 23.9), and IgG4 (average: 35.5, SD ± 18.2). We performed IHC staining for NELL-1 in 126 cases of PLA2R-negative MN from the pilot and discovery cohorts. Twenty-nine cases (23.0%) were positive for NELL-1 (6 in the pilot and 23 in the discovery cohort). All 29 positive cases showed bright (2–3+/3) granular staining for NELL-1 along the GBM. Importantly, there was no significant mesangial staining. NELL-1 staining in 6 cases is shown in Figure 3a. Segmental granular capillary wall staining for NELL-1 was seen in 6 cases (20.6%). Review of electron microscopy confirmed the segmental subepithelial deposits in all 6 cases (Supplementary Figure S2). There was no staining along the Bowman’s capsule, tubular basement membranes, or in vessel walls. The positive NELL-1 granular staining mirrored the granular IgG along the GBM seen in each case. All control cases were negative for NELL-1. Representative negative staining for NELL-1 in PLA2R-associated MN, focal segmental glomerulosclerosis, IgA nephropathy, and diabetes is shown in Figure 3b. Representative NELL-1 staining in the remaining NELL-1–positive MN is also shown in the Supplementary Figure S3.Figure 3Immunohistochemical (IHC) stain for neural epidermal growth factor-like 1 protein (NELL-1) in NELL-1–associated membranous nephropathy (MN), M-type phospholipase A2 receptor (PLA2R)–associated MN, and control cases. (a) The bright granular capillary wall staining for NELL-1 along the glomerular basement membranes in 6 cases of NELL-1–associated MN are shown. Note the segmental capillary wall staining in case 9. (b) The negative NELL-1 staining in control cases are shown. There was no capillary wall staining for NELL-1 in (A,B) 2 cases of PLA2R-associated MN, (C) an additional case that was PLA2R-negative but also NELL-1–negative, (D) IgA nephropathy, (E) focal segmental glomerulosclerosis (FSGS), and (F) diabetic glomerulus. Note very weak podocyte staining for NELL-1 but negative capillary wall staining. (c) Immunofluorescence (IF) and IHC show bright capillary wall staining for NELL-1 of 3 cases of the French validation cohort. (A,B) Case 1 (patient 30) was stained by both IHC and IF, and (C,D) the remaining 2 cases (patients 31 and 32) were stained with IF only. (d) IF shows capillary wall staining for NELL-1 of (A,B) 2 cases of the Belgian validation cohort. Bars = 20 μm. To optimize viewing of this image, please see the online version of this article at www.kidney-international.org.View Large Image Figure ViewerDownload Hi-res image Download (PPT) Five of 90 cases (5.9%) of PLA2R- and THSD7A-negative MN were positive for NELL-1 staining in the validation cohorts. Three of 45 cases (6.7%) were positive for NELL-1 staining. The first positive case (patient 30) was detected at the Mayo Clinic and the remaining 2 cases (patients 31 and 32) at Tenon Hospital. Both IHC and immunofluorescence (IF) studies for NELL-1 were done in the first case (patient 30), while IF studies were done for NELL-1 detection in the other 2 cases (patients 31 and 32) (Figure 3c). Two of 39 cases (5.1%) (patients 33 and 34) were positive for NELL-1 staining by IF staining (Figure 3d). Those cases were recruited at Cliniques universitaires Saint-Luc in Brussels and detected at Tenon Hospital. We performed confocal IF microscopy to show that the NELL-1 and IgG colocalize along the GBM (Figure 4). Superimposition of the 2 signals (yellow, Figure 4c and f) and laser quantitative analysis (Figure 4g) confirm the colocalization of NELL-1 and IgG, further corroborating that the subepithelial deposits contain both NELL-1 and IgG. A second case is shown in Supplementary Figure S4. Western blot analyses were performed using recombinant human NELL-1 to determine the presence of circulating anti-NELL-1 antibodies in the serum of 5 patients—4 patients from the validation cohort and 1 from the discovery cohort. All 5 patients showed reactivity against NELL-1 under nonreducing conditions (patients are labeled as MN in Table 1); NELL-1 was detected as a 280-kDa homodimer and a 420-kDa homotrimer. Furthermore, sera were available at different points in patient MN2. The MN2 sera were tested both prior to and during follow-up (Figure 5a). Sera from patients with PLA2R-associated MN, minimal change disease, and IgA nephropathy did not show any reactivity against NELL-1. There was no reactivity under reducing conditions where NELL-1 resolves as monomeric bands of about 130 kDa, suggesting that NELL-1 autoantibody recognizes conformation-dependent epitopes.Table 1Laboratory and kidney biopsy findings of NELL-1–associated MNCaseAgeSexUrinary protein g/24 haMean urinary protein 7 ± 3.7 g/24 h.Serum creatinine mg/dlbMean serum creatinine 1.7 ± 1.3 mg/dl; cases 30, 31, 32, and 34 were associated with malignancies.Sclerosed/total glomeruliIFTAIFEM1 (MN5)cMN1 to the 5 cases in which serum was available and Western blot studies were performed Figure to the 5 cases in which serum was available and Western blot studies were performed Figure to the 5 cases in which serum was available and Western blot studies were performed Figure to the 5 cases in which serum was available and Western blot studies were performed Figure to the 5 cases in which serum was available and Western blot studies were performed Figure electron IF, immunofluorescence microscopy of out of and tubular MN, membranous not not NELL-1, neural epidermal growth factor-like 1 segmental to urinary protein 7 ± 3.7 g/24 serum creatinine 1.7 ± 1.3 mg/dl; cases 30, 31, 32, and 34 were associated with to the 5 cases in which serum was available and Western blot studies were performed Figure in a electron IF, immunofluorescence microscopy of out of and tubular MN, membranous not not NELL-1, neural epidermal growth factor-like 1 segmental to we also characterized the NELL-1 and showed that the IgG is IgG1 in patients and and IgG2 and IgG4 were also along with IgG1 in patient MN2 (Figure We identified 29 cases of NELL-1–associated MN from the pilot and discovery cohorts (patients There were patients and 14 patients The at was 63.1 ± The serum creatinine and at was 1.7 ± and ± g/24 respectively. urinary protein was not done in 7 the of 1 patient with positive including were The kidney biopsy of all cases of NELL-1–associated MN showed the findings of GBM on microscopy, bright IgG and 3 staining along the capillary wall on IF microscopy, and subepithelial deposits on electron an average of glomeruli were of which ± were IF microscopy showed bright staining for IgG (2–3+/3) and in all cases. 1 case showed and 2 cases showed The remaining cases were negative for and All cases showed staining for (2–3+/3) and (2–3+/3) IF staining for PLA2R was negative in all cases. microscopy showed subepithelial deposits in all cases, and in 6 cases, the subepithelial deposits were in a segmental but not all the capillary and mesangial deposits were not were not The 3 positive NELL-1 cases of the French validation cohort were also and 1 (patient 30) (patient and the (patient 32) In all 3 patients of that was at the time or a the of MN. (patient of the 2 positive NELL-1 cases of the Belgian validation cohort was while the other (patient 34) was an patient 34 of MN. The and findings are shown in Table MN is the most of in is by against target antigens in the GBM and of on the of the target MN is also as PLA2R-positive and negative MN. In the negative cases, the target antigen(s) elusive. We identified 2 novel and in patients with MN associated with S. Madden B.J. Debiec H. et al.Exostosin 1/exostosin 2–associated membranous nephropathy.J Am Soc Nephrol. 2019; 30: 1123-1136Crossref PubMed Scopus (107) Google Scholar We used a of laser and IHC to identify and We that we could identify the antigen(s) of the remaining to of primary PLA2R- and THSD7A-negative MN cases using the microdissection with MS/MS for the of a of proteins along with of using total spectral In this we were to identify to glomerular proteins per most of which are with total spectral we that PLA2R was the most abundant protein in PLA2R-associated MN with the S. Madden B.J. Debiec H. et al.Exostosin 1/exostosin 2–associated membranous nephropathy.J Am Soc Nephrol. 2019; 30: 1123-1136Crossref PubMed Scopus (107) Google Scholar Here, we detected a novel protein, NELL-1, in glomeruli dissected from PLA2R-negative MN. The NELL-1 spectral counts were the and were comparable to the and basement membrane proteins and were similar to the counts of PLA2R and in PLA2R- and EXT1/EXT2-associated MN. The high NELL-1 spectral count was also by the sequence coverage for the NELL-1 There were NELL-1 spectral counts in PLA2R-positive MN and control cases. IHC confirmed MS/MS and bright granular capillary wall NELL-1 staining that was the GBM and mirrored the IgG staining. There was of MS/MS and that all NELL-1–positive MN cases detected on MS/MS (pilot cohort) were positive on IHC and all NELL-1–positive MN cases detected on IHC (discovery cohort) were also positive for NELL-1 on there was segmental positive IHC staining for NELL-1 in a electron microscopy in cases also showed segmental subepithelial The of NELL-1 staining along the GBM and with the subepithelial deposits that this protein is from the circulating antigens or immune is that NELL-1 is from mesangial or there was no mesangial or staining in the NELL-1–positive MN. Furthermore, IF confocal microscopy studies showed that both NELL-1 and IgG to the that NELL-1 is the target antigen for the IgG. was further confirmed by of circulating antibodies to NELL-1 in the sera of 5 In 1 patient in sera was available at different time reactivity to NELL-1 at the time of with and that the patient was but the of is in with results seen in PLA2R-associated MN where of PLA2R L.H. Beck et of in membranous nephropathy.J Am Soc Nephrol. PubMed Scopus Google Scholar that NELL-1 can be used for follow-up to be confirmed in further et for a to membranous nephropathy.J Am Soc Nephrol. 2017; PubMed Scopus Google Scholar NELL-1–associated MN be to the of MN, including PLA2R- and MN. is seen in has no has no IgG4 with and there is an of features as for on in validation disease, and IgG performed in cases of NELL-1–positive MN by IF and MS/MS showed that the was the IgG in PLA2R-positive MN is to be The kidney biopsy also shows no features that to a as features on microscopy, Ig staining including on IF microscopy, and in and mesangial or deposits on electron NELL-1 is a to a that is in neural tissue encoding a protein with EGF-like S. S. E. et encoding a protein with EGF-like repeats is in neural of PubMed Scopus Google Scholar expression is in all and the expression was in the as and kidney very of S. S. E. et encoding a protein with EGF-like repeats is in neural of PubMed Scopus Google Scholar 2 to NELL-1, and been et and of two novel human and encoding proteins with six EGF-like PubMed Scopus (109) Google Scholar The of NELL-1 and are about suggesting that the 2 proteins are distinct with different The and are in and in the PubMed Scopus Google Scholar NELL-1 a protein of and including a 4 and 6 EGF-like repeats (Figure et of NELL-1, a growth factor associated with in PubMed Scopus Google Scholar The is the and the repeats are the protein S. et and expression analysis of neural proteins and PubMed Scopus Google Scholar NELL-1 is in and The of NELL-1 H. et and of recombinant human protein in human kidney PubMed Scopus Google H. et of PubMed Scopus Google Scholar NELL-1 is in patients with of the where is H. et NELL-1 in PubMed Scopus Google et of a in during PubMed Scopus Google Scholar NELL-1 also the S. et in Clin Invest. PubMed Scopus Google Scholar In the NELL-1 expression is in while is in the glomeruli 5% to of glomerular NELL-1 at the et and of two novel human and encoding proteins with six EGF-like PubMed Scopus (109) Google Scholar human kidney were on NELL-1 was in the of the suggesting that NELL-1 is as an and be in the H. et and of recombinant human protein in human kidney PubMed Scopus Google Scholar was shown NELL-1 expression is in of while is in and is to an in the et and on of and in human 2015; PubMed Scopus Google Scholar To the of of NELL-1 has not been in any kidney NELL-1–associated MN to be unique kidney associated with of NELL-1. is the case for PLA2R and S. et al.Thrombospondin type 1 7A to the and membrane of Am Soc Nephrol. 2019; 30: PubMed Scopus Google Scholar anti-NELL-1 antibodies a conformation-dependent both in 280-kDa and 420-kDa a studies are to determine NELL-1 at the podocyte and the of anti-NELL-1 antibodies in podocyte and a of 126 of NELL-1–positive MN were detected in the cohort cases at the Mayo Clinic with the of cases of from the French and Belgian validation cohorts. be to as is seen in the of PLA2R- and S. et A2 receptor (PLA2R) and glomerular PLA2R expression in patients with membranous 2016; PubMed Scopus Google Scholar Thus, with to a study showed of PLA2R-associated MN at the Tenon et of PLA2R detected by of antibodies with detection of PLA2R antigen in membranous nephropathy: a study 14 2017; PubMed Scopus Google Scholar while there was a of of MN at the Mayo Clinic a study on MN a of the patients from 6 C. et for thrombospondin type 1 domain-containing 7A in membranous 2019; Full Text Full Text PDF PubMed Scopus Google Scholar while 1 of patients in Membranous of et or in the of membranous nephropathy.N Engl J Med. 2019; PubMed Scopus Google Scholar and 2 of patients at Mayo Clinic and to be THSD7A positive was detected in 4 of 5 NELL-1–positive cases of the validation while of the 29 NELL-1 of the pilot and discovery Mayo Clinic cohort cases The findings that NELL-1–associated MN can in different including as is also the case for PLA2R-associated MN. also the whether the subset of NELL-1–positive MN associated with can be as primary MN. studies are to this 34 cases were identified in PLA2R-negative with all cohorts and that is the of THSD7A cases in the study of N.M. Beck L.H. Meyer-Schwesinger C. et al.Thrombospondin type-1 domain-containing 7A in idiopathic membranous nephropathy.N Engl J Med. 2014; 371: 2277-2287Crossref PubMed Scopus (496) Google Scholar the findings that NELL-1 is the second most antigen in primary MN. studies are to the of NELL-1–positive MN. In we identified a novel protein NELL-1 in a subset of PLA2R-negative MN in NELL-1–associated MN to be a distinct type of primary MN. were in the of and Mayo for and and microscopy, IF microscopy including PLA2R and electron microscopy was performed in each case of MN. The was from the The study was by the Mayo Clinic Review control cases, we performed MS/MS on cases that cases of time kidney cases of minimal change disease, cases of focal segmental glomerulosclerosis, 7 cases of diabetic glomerulosclerosis, 5 cases of IgA nephropathy, and 23 cases of PLA2R-associated MN. control we used 20 cases that 4 cases of focal segmental glomerulosclerosis, 4 cases of IgA nephropathy, 4 cases of and cases of PLA2R-associated MN. Two validation cohorts were kidney biopsy of tissue were by the et and analyzed in the by IHC for NELL-1. All 23 cases were PLA2R- and THSD7A-negative MN. An additional cases of PLA2R- and THSD7A-negative MN were stained for NELL-1 at using IF kidney biopsy of tissue of PLA2R- and THSD7A-negative MN were in the cases were stained for NELL-1 at using each case were and on a membrane laser microdissection and using a the glomeruli were to approximately to per case. were with and for MS/MS The were identified by tandem MS/MS using a Mass to a All MS/MS samples were analyzed using and to a human was used to and protein were at by the with protein a 2 and a using E. et for proteins by tandem mass PubMed Scopus Google Scholar of laser and MS/MS are in the Supplementary and Supplementary Table and IHC staining were performed at the using the were at 5 and IHC staining was performed for the NELL-1 stain were for 20 using 2 and in for 5 The NELL-1 primary was to in and for The detection used was the primary and and was by in and from the To this were with were for 5 using and followed by in and this is not the with the the was were from the and in for 5 were in of and in 3 of prior to in IF staining was performed on for using target high in The NELL-1 primary was to in serum and and at 4 with biopsy Next, the were with IgG IgG was then with the tissue as were in and with of NELL-1 and IgG along the glomerular basement membrane was by confocal microscopy using a and analyzed with The protein recombinant human NELL-1 was with nonreducing or reducing and for 5 were to and in were to membranes to and the membranes were with were at 4 with sera from controls and antibodies against NELL-1 were and for 2 at with human or IgG, proteins were with detection in nonreducing were with IgG antibodies then with IgG All the no We to the Mayo of the Mayo Clinic of and and the Mayo and the the of the for in and the We are to the of the patients in the of and at Tenon the 20 to from the of and to for in Western We to and all of the Tenon for in and We also the from the for from the of of the Cliniques universitaires for and for and the the the kidney IHC and and performed the laser microdissection and in performed the IHC. and tissue for the validation cohort and also performed the confocal studies and Western blot and and tissue for the Belgian validation cohort. The was and by with as from the of the Download with Supplementary and membranous nephropathy: a of nephropathy (MN) is primary or associated with each with unique glomerular The discovery of the phospholipase A2 receptor (PLA2R) antigen in primary MN of MN and to major antigens in MN the THSD7A antigen and et identified a NELL-1, in primary MN, the of patients antigen unknown. PDF In this and a novel of that kidney followed by kidney In this was followed by a of the kidney significant tubular kidney tubular and of the at followed by a PDF

OBJECTIVES: To refine the Family Satisfaction in the Intensive Care Unit (FS-ICU) survey and develop a validated method for scoring the instrument. DESIGN: Instrument development study, using data from two prospective cohort studies. SETTING: Intensive care units in seven university-affiliated hospitals (six Canadian, one United States). SUBJECTS: Family members of ICU patients. INTERVENTIONS: Based on a priori criteria, items were tagged for potential removal and discussed with the FS-ICU developers. Factor analysis was used to test the conceptual structure of the instrument and develop a scoring method based on scales and subscales. The new scoring method was validated in the U.S. cohort using the Quality of Dying and Death (QODD) instrument and nurse-assessed quality indicators. MEASUREMENTS AND MAIN RESULTS: A total of 1,038 family members completed the FS-ICU across seven sites. Fifteen items were initially tagged for possible removal. After consensus with the developers, ten items were dropped (and 24 were retained in the final instrument). Factor analysis explained 61.3% of the total variance using a two-factor model. The first factor pertained to satisfaction with care (14 items). The second factor encompassed satisfaction with decision making (10 items). A scoring method was developed based on this conceptual model. In validity testing, the FS-ICU was significantly correlated with the Family-QODD total score (Spearman's .56, p < .001) as well as individual QODD items such as quality of care by all providers (.64, p < .001). The FS-ICU also correlated significantly with multiple nurse-assessed quality indicators. CONCLUSIONS: The shortened FS-ICU measures two main conceptual domains-satisfaction with care and satisfaction with decision making. Scores on the FS-ICU show good validity against other indicators of ICU quality. The instrument holds promise as a useful outcome measure in studies that attempt to improve this component of ICU care.

Acidosis, a common characteristic of the tumor microenvironment, is associated with alterations in metabolic preferences of cancer cells and progression of the disease. Here we identify the TGF-β2 isoform at the interface between these observations. We document that acidic pH promotes autocrine TGF-β2 signaling, which in turn favors the formation of lipid droplets (LD) that represent energy stores readily available to support anoikis resistance and cancer cell invasiveness. We find that, in cancer cells of various origins, acidosis-induced TGF-β2 activation promotes both partial epithelial-to-mesenchymal transition (EMT) and fatty acid metabolism, the latter supporting Smad2 acetylation. We show that upon TGF-β2 stimulation, PKC-zeta-mediated translocation of CD36 facilitates the uptake of fatty acids that are either stored as triglycerides in LD through DGAT1 or oxidized to generate ATP to fulfill immediate cellular needs. We also address how, by preventing fatty acid mobilization from LD, distant metastatic spreading may be inhibited.

Dendritic cells (DCs) initiate adaptive immune responses in lymph nodes (LNs). In mice, LN DCs can be divided into resident and tissue-derived populations, the latter of which migrate from the peripheral tissues. In humans, different subsets of DCs have been identified in the blood, spleen, and skin, but less is known about populations of resident and migratory tissue-derived DCs in LNs. We have analyzed DCs in human LNs and identified two populations of resident DCs that are present in all LNs analyzed, as well as in the spleen and tonsil, and correspond to the two known blood DC subtypes. We also identify three main populations of skin-derived migratory DCs that are present only in skin-draining LNs and correspond to the DC subsets found in the skin. Resident DCs subsets induce both Th1 and Th2 cytokines in naive allogeneic T lymphocytes, whereas the corresponding blood subsets failed to induce efficient Th2 polarization. LN-resident DCs also cross-present antigen without in vitro activation, whereas blood DCs fail to do so. Among migratory DCs, one subset was poor at both CD4(+) and CD8(+) T cell activation, whereas the other subsets induced only Th2 polarization. We conclude that in humans, skin-draining LNs host both resident and migratory DC subsets with distinct functional abilities.

Exosomes are secreted vesicles formed in late endocytic compartments. Mature dendritic cells (DCs) secrete exosomes bearing functional MHC-peptide complexes and high levels of ICAM-1. Such exosomes can activate Ag-specific naive T cells but only after recapture by recipient APCs. In this study, we addressed the molecular mechanisms of interaction between exosomes and recipient DCs. We show that exosomes can be presented by mouse DCs without the need for internalization and processing. Exosomes interact with DCs through a specific saturable receptor. Although the two major ligands of ICAM-1, LFA-1 and Mac-1, are expressed by lymphoid organ DCs, only LFA-1 is required for exosome capture by these cells. Accordingly, we show that CD8(+) DCs express higher levels of LFA-1 than CD8(-) DCs, and that they are the main recipients of exosomes in vivo. We propose a new role for LFA-1 on DCs, as a receptor for exosomes to favor Ag transfer between DCs in vivo.

Abstract. Since 2005 the European Flood Alert System (EFAS) has been producing probabilistic hydrological forecasts in pre-operational mode at the Joint Research Centre (JRC) of the European Commission. EFAS aims at increasing preparedness for floods in trans-national European river basins by providing medium-range deterministic and probabilistic flood forecasting information, from 3 to 10 days in advance, to national hydro-meteorological services. This paper is Part 2 of a study presenting the development and skill assessment of EFAS. In Part 1, the scientific approach adopted in the development of the system has been presented, as well as its basic principles and forecast products. In the present article, two years of existing operational EFAS forecasts are statistically assessed and the skill of EFAS forecasts is analysed with several skill scores. The analysis is based on the comparison of threshold exceedances between proxy-observed and forecasted discharges. Skill is assessed both with and without taking into account the persistence of the forecasted signal during consecutive forecasts. Skill assessment approaches are mostly adopted from meteorology and the analysis also compares probabilistic and deterministic aspects of EFAS. Furthermore, the utility of different skill scores is discussed and their strengths and shortcomings illustrated. The analysis shows the benefit of incorporating past forecasts in the probability analysis, for medium-range forecasts, which effectively increases the skill of the forecasts.

This is the first completed prospective randomized clinical efficacy trial of antifungals in the treatment of invasive aspergillosis (IA) and the first to compare the clinical efficacy of two dosages of liposomal amphotericin B (L-AmB) for IA in neutropenic patients with cancer or those undergoing bone marrow transplantation. Eighty-seven of 120 patients were eligible and evaluable. Clinical responses were documented for 26 (64%) of 41 patients receiving 1 mg/(kg.d) (L-AmB-1) and 22 (48%) of 46 receiving 4 mg/(kg.d) (L-AmB-4). Radiologic response rates were similar: 24 (58%) of the L-AmB-1 recipients and 24(52%) of the L-AmB-4 recipients. The six-month survival rates were 43% (L-AmB-1) and 37% (L-AmB-4). These differences were not significant. The numbers of deaths directly due to IA at 6 months were similar: 9 (22%) of 41 L-AmB-1 recipients and 9 (20%) of 46 L-AmB-4 recipients. No other variable independently influenced survival, apart from central nervous system IA. L-AmB is effective in treating approximately 50%-60% of patients who have IA. A 1-mg/(kg.d) dosage is as effective as a 4-mg/(kg.d) dosage, and no advantages to use of the higher, more expensive, dosage has been observed.

This review presents an overview of analytical methods for the analysis of pesticide residues in grapes and by-products in the last decade. The most widely used detection technique for the determination of pesticides in grapes is mass spectrometry combined with gas and/or liquid chromatography. In general, multi-residue methods with selective sample treatment methodologies have been developed for this purpose. However, this review focuses not only on these common multi-residue methods but also on specific methodologies as single-residue methods for the analysis of pesticides in grapes and by-products. Finally, the limitations of multi-residue methods, the future perspectives and the trends for pesticide residue analysis in grapes are reviewed.

A method is described for isolating mutants of Saccharomyces cerevisiae which have lost repressibility by exogenous arginine for ornithine transcarbamylase. Besides permeability mutants, three complementary classes of mutations were found: argRI, argRII and argRIII which are recessive and define three loci. No evidence for a linkage between any of these three loci or with the gene coding for ornithine transcarbamylase has been obtained. Strains bearing mutations at either of these loci cannot be distinguished on a phenotype basis: after growth on minimal medium, the l ‐ornithine carbamoyl transferase activity is twice that of the wild type strain; the mutations modify neither the growth rate nor the permeability to arginine. The mutations might affect the structure of an hetero‐polypeptidic aporepressor. The level of ornithine transcarbamylase in diploids is proportional to the number of argF + genes in regulated as well as in non‐regulated cells.

Developmental neurotoxicity (DNT) and many forms of reproductive toxicity (RT) often manifest themselves in functional deficits that are not necessarily based on cell death, but rather on minor changes relating to cell differentiation or communication. The fields of DNT/RT would greatly benefit from in vitro tests that allow the identification of toxicant-induced changes of the cellular proteostasis, or of its underlying transcriptome network. Therefore, the 'human embryonic stem cell (hESC)-derived novel alternative test systems (ESNATS)' European commission research project established RT tests based on defined differentiation protocols of hESC and their progeny. Valproic acid (VPA) and methylmercury (MeHg) were used as positive control compounds to address the following fundamental questions: (1) Does transcriptome analysis allow discrimination of the two compounds? (2) How does analysis of enriched transcription factor binding sites (TFBS) and of individual probe sets (PS) distinguish between test systems? (3) Can batch effects be controlled? (4) How many DNA microarrays are needed? (5) Is the highest non-cytotoxic concentration optimal and relevant for the study of transcriptome changes? VPA triggered vast transcriptional changes, whereas MeHg altered fewer transcripts. To attenuate batch effects, analysis has been focused on the 500 PS with highest variability. The test systems differed significantly in their responses (<20 % overlap). Moreover, within one test system, little overlap between the PS changed by the two compounds has been observed. However, using TFBS enrichment, a relatively large 'common response' to VPA and MeHg could be distinguished from 'compound-specific' responses. In conclusion, the ESNATS assay battery allows classification of human DNT/RT toxicants on the basis of their transcriptome profiles.

OBJECTIVES: The present study is an attempt to experimentally induce a younger subjective age among older adults and to test whether they show better physical functioning when they are induced to feel younger. METHOD: Participants were 49 older adults aged between 52 and 91 years. Following an initial measure of handgrip performance as an indicator of physical functioning, participants in the experimental condition received positive feedback regarding their performance compared with their same-aged peers, whereas participants in the control condition did not receive any information. Participants in both groups then completed a second handgrip measure. Subjective age was assessed before the initial handgrip task and after the experimental manipulation. RESULTS: Participants in the experimental group felt younger than their age and showed a significant increase in grip strength, whereas no changes in subjective age and grip strength were observed in the control condition. DISCUSSION: This study is among the first to induce a younger subjective age. It supports the notion that redirecting older adults' attention to downward social comparison with same-aged peers is a promising strategy to maintain a sense of feeling younger. In addition, our results provide an initial positive answer to the question of whether feeling younger translates into better physical functioning.

BACKGROUND: Most Clostridium difficile infection (CDI) surveillance programs neither specify the diagnostic method to be used nor stratify rates accordingly. We assessed the difference in healthcare-associated CDI (HA-CDI) incidence and complication rates obtained by 2 validated diagnostic methods. METHODS: This was a prospective cohort study of patients for whom a C. difficile test was ordered between 1 August 2010 and 31 July 2011. All specimens were tested in parallel by a commercial polymerase chain reaction (PCR) assay targeting toxin B gene tcdB, and a 3-step algorithm detecting glutamate dehydrogenase and toxins A and B by enzyme immunoassay and cell culture cytotoxicity assay (EIA/CCA). CDI incidence rate ratios were calculated using univariate Poisson regression. RESULTS: A total of 1321 stool samples were tested during a period totaling 95 750 patient-days. Eighty-five HA-CDI cases were detected by PCR and 56 cases by EIA/CCA (P = .01). The overall incidence rate was 8.9 per 10 000 patient-days (95% confidence interval [CI], 7.1-10.9) by PCR and 5.8 per 10 000 patient-days (95% CI, 4.4-7.4) by EIA/CCA (P = .01). The incidence rate ratio comparing PCR and EIA/CCA was 1.52 (95% CI, 1.08-2.13; P = .015). Overall complication rate was 27% (23/85) when CDI was diagnosed by PCR and 39% (22/56) by EIA/CCA (P = .16). Cases detected by PCR only were less likely to develop a complication of CDI compared with cases detected by both PCR and EIA/CCA (3% vs 39%, respectively; P < .001). CONCLUSIONS: Performing PCR instead of EIA/CCA is associated with a >50% increase in the CDI incidence rate. Standardization of diagnostic methods may be indicated to improve interhospital comparison.

OBJECTIVES: To investigate cancer patients' desire for psychological support and to identify patients' sociodemographic, disease-related and psychological factors associated with this desire. METHODS: The study is part of a multicenter, cross-sectional study assessing cancer patients' needs and desire for psychological support. Patients completed the Hospital Anxiety and Depression Scale, the Ways of Coping Checklist, the Cancer Rehabilitation Evaluation System and reported their desire for psychological support. RESULTS: Among the 381 included patients, women (26%) desired psychological support significantly more often than men (11%) (p<0.001). Patients' desire for psychological support was associated with being younger (OR=0.94; p<0.001 for women and OR=0.93; p=0.007 for men) and having a support-seeking coping (OR=1.10; p=0.010 for women and OR=1.36; p=0.003 for men). Other contextual factors such as difficulties encountered and treatment modalities were diversely associated with women and men's desire for psychological support. Neither women's, nor men's psychological distress was associated with their desire for psychological support. CONCLUSIONS: One female cancer patient out of four and one male cancer patient out of ten desire psychological support. Results emphasize the need to screen not only for cancer patients' distress but also for their desire for psychological support. This will allow implementing psychological interventions according to patients' needs and desire.