Defining Chlorophyll-a Reference Conditions in European Lakes

The concept of “reference conditions” describes the benchmark against which current conditions are compared when assessing the status of water bodies. In this paper we focus on the establishment of reference conditions for European lakes according to a phytoplankton biomass indicator—the concentration of chlorophyll-a. A mostly spatial approach (selection of existing lakes with no or minor human impact) was used to set the reference conditions for chlorophyll-a values, supplemented by historical data, paleolimnological investigations and modelling. The work resulted in definition of reference conditions and the boundary between “high” and “good” status for 15 main lake types and five ecoregions of Europe: Alpine, Atlantic, Central/Baltic, Mediterranean, and Northern. Additionally, empirical models were developed for estimating site-specific reference chlorophyll-a concentrations from a set of potential predictor variables. The results were recently formulated into the EU legislation, marking the first attempt in international water policy to move from chemical quality standards to ecological quality targets

Tolerability and safety of intravitreal aflibercept 8 mg in the Phase 3 PULSAR trial of patients with neovascular age-related macular degeneration

Purpose : In PULSAR (NCT04423718), the primary endpoint of change from baseline in best-corrected visual acuity at Week 48 (non-inferiority margin at 4 letters) was met with intravitreal aflibercept 8 mg every 12 or 16 weeks (8q12 or 8q16) vs aflibercept 2 mg every 8 weeks (2q8). Here, we report safety and tolerability through Week 48. Methods : PULSAR is an ongoing, double-masked, 96-week, Phase 3 trial. Patients aged ≥50 years with treatment-naïve neovascular age-related macular degeneration (nAMD) were randomly assigned 1:1:1 to 8q12, 8q16 or 2q8, each after three initial monthly injections. Results : Of 1,009 patients (mean age 74.5 years) randomly assigned and treated (2q8: n=336; 8q12: n=335; 8q16: n=338), 11 (1.1%) discontinued with adverse events (AEs) as the primary reason. In the 2q8, 8q12 and 8q16 groups, 46/336 (13.7%), 34/335 (10.1%), and 32/338 patients (9.5%) had treatment-emergent (TE) non-ocular serious AEs, respectively; adjudicated Antiplatelet Trialists’ Collaboration (APTC) events occurred in 1.5%, 0.3%, and 0.6%, respectively. In total, 38.3% of patients experienced at least one TE ocular AE in the study eye (2q8: 130/336 [38.7%]; 8q12: 129/335 [38.5%]; 8q16: 127/338 patients [37.6%]); most common were reduced visual acuity (6.0%, 3.6%, 5.3% for 2q8, 8q12 and 8q16, respectively), cataracts (3.0%, 3.6%, and 3.6%, respectively), and retinal hemorrhage (4.2%, 3.3%, and 3.0%, respectively). Pre-injection intraocular pressure (IOP) values were similar to baseline at all timepoints through Week 48. The proportion of patients with pre- or post-dose IOP ≥35 mmHg was similar in 2q8, 8q12 and 8q16 groups (0.3%, 0.9%, and 0.3%, respectively). Seven patients had intraocular inflammation (IOI; 2q8: 0.6%; 8q12: 1.2%; 8q16: 0.3%), none discontinued and there were no cases of endophthalmitis or occlusive retinal vasculitis. In all groups, proportions of patients with TE anti-drug antibody (ADA)-positive status were very low and consistent with rates previously reported for aflibercept 2 mg. Conclusions : The safety profile of aflibercept 8 mg was similar to that of aflibercept 2 mg through Week 48 in patients with nAMD. Of note, rates of APTC events, IOI and TE ADA-positivity were very low; IOI was not associated with positive ADA status. This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field