Search

The ongoing threat of COVID-19 has highlighted the need for effective prophylaxis and convenient therapies, especially for outpatient settings. We have previously developed highly potent single-domain (VHH) antibodies, also known as nanobodies, that target the Receptor Binding Domain (RBD) of the SARS-CoV-2 Spike protein and neutralize the Wuhan strain of the virus. In this study, we present a new generation of anti-RBD nanobodies with superior properties. The primary representative of this group, Re32D03, neutralizes Alpha to Delta as well as Omicron BA.2.75; other members neutralize, in addition, Omicron BA.1, BA.2, BA.4/5, and XBB.1. Crystal structures of RBD-nanobody complexes reveal how ACE2-binding is blocked and also explain the nanobodies' tolerance to immune escape mutations. Through the cryo-EM structure of the Ma16B06-BA.1 Spike complex, we demonstrated how a single nanobody molecule can neutralize a trimeric spike. We also describe a method for large-scale production of these nanobodies in Pichia pastoris, and for formulating them into aerosols. Exposing hamsters to these aerosols, before or even 24 h after infection with SARS-CoV-2, significantly reduced virus load, weight loss and pathogenicity. These results show the potential of aerosolized nanobodies for prophylaxis and therapy of coronavirus infections., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:The following authors are employes of Scinai Immunotherapeutics Ltd., a company dedicated to developing Nanobodies for clinical use. Tamar Ben-Yedidia, Oded Ovadia, Dalit Weinstein Fischer, Amir Reichman. Moreover, Matthias Dobbelstein is a member of the scientific advisory board of Scinai Immunotherapeutics Ltd.., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Triclosan (TCS) is a broad-spectrum antimicrobial used in consumer products including toothpaste and hand soap. After being used, TCS is washed or rinsed off and residuals that are not biodegraded or otherwise removed during wastewater treatment can enter the aquatic environment in wastewater effluents and sludges. The environmental exposure and toxicity of TCS has been the subject of various scientific and regulatory discussions in recent years. There have been a number of publications in the past 5 y reporting toxicity, fate and transport, and in-stream monitoring data as well as predictions from aquatic risk assessments. State-of-the-science probabilistic exposure models, including Geography-referenced Regional Exposure Assessment Tool for European Rivers (GREAT-ER) for European surface waters and Pharmaceutical Assessment and Transport Evalutation (PhATE) for US surface waters, have been used to predict in-stream concentrations (PECs). These models take into account spatial and temporal variability in river flows and wastewater emissions based on empirically derived estimates of chemical removal in wastewater treatment and in receiving waters. These model simulations (based on realistic use levels of TCS) have been validated with river monitoring data in areas known to be receiving high wastewater loads. The results suggest that 90th percentile (low flow) TCS concentrations are less than 200 ng/L for the Aire-Calder catchment in the United Kingdom and between 250 ng/L (with in-stream removal) and 850 ng/L (without in-stream removal) for a range of US surface waters. To better identify the aquatic risk of TCS, a species sensitivity distribution (SSD) was constructed based on chronic toxicity values, either no observed effect concentrations (NOECs) or various percentile adverse effect concentrations (EC10-25 values) for 14 aquatic species including fish, invertebrates, macrophytes, and algae. The SSD approach is believed to represent a more realistic threshold of effect than a predicted no effect concentration (PNEC) based on the data from the single most sensitive species tested. The log-logistic SSD was used to estimate a PNEC, based on an HC5,50 (the concentration estimated to affect the survival, reproduction and/or growth of 5% of species with a 50% confidence interval). The PNEC for TCS was 1,550 ng/L. Comparing the SSD-based PNEC with the PECs derived from GREATER and PhATE modeling to simulate in-river conditions in Europe and the United States, the PEC to PNEC ratios are less than unity suggesting risks to pelagic species are low even under the highest likely exposures which would occur immediately downstream of wastewater treatment plant (WWTP) discharge points. In-stream sorption, biodegradation, and photodegradation will further reduce pelagic exposures of TCS. Monitoring data in Europe and the United States corroborate the modeled PEC estimates and reductions in TCS concentrations with distance downstream of WWTP discharges. Environmental metabolites, bioaccumulation, biochemical responses including endocrine-related effects, and community level effects are far less well studied for this chemical but are addressed in the discussion. The aquatic risk assessment for TCS should be refined as additional information becomes available.