Lyophilisation of influenza, rabies and Marburg lentiviral pseudotype viruses for the development and distribution of a neutralisation-assay based diagnostic kit

Pseudotype viruses (PVs) are chimeric, replication-deficient virions that mimic wild-type virus entry mechanisms and can be safely employed in neutralisation assays, bypassing the need for high biosafety requirements and performing comparably to established serological assays. However, PV supernatant necessitates -80°C long-term storage and cold-chain maintenance during transport, which limits the scope of dissemination and application throughout resource-limited laboratories. We therefore investigated the effects of lyophilisation on influenza, rabies and Marburg PV stability, with a view to developing a pseudotype virus neutralisation assay (PVNA) based kit suitable for affordable global distribution. Infectivity of each PV was calculated after lyophilisation and immediate reconstitution, as well as subsequent to incubation of freeze-dried pellets at varying temperatures, humidities and timepoints. Integrity of glycoprotein structure following treatment was also assessed by employing lyophilised PVs in downstream PVNAs. In the presence of 0.5M sucrose-PBS cryoprotectant, each freeze-dried pseudotype was stably stored for 4 weeks at up to 37°C and could be neutralised to the same potency as unlyophilised PVs when employed in PVNAs. These results confirm the viability of a freeze-dried PVNA-based kit, which could significantly facilitate low-cost serology for a wide portfolio of emerging infectious viruses.

Sustainable freight: impacts of the London congestion charge and low emissions zone

This paper assesses whether two sustainability policies currently in effect in London, a congestion charge zone and a low emission zone, have affected freight operations and reduced vehicle kilometers travelled. It investigates responses by freight operators, including re-timing, re-routing, or reducing the number of trips, or replacing vehicles. Freight traffic trends from 1994 to 2012 were identified using road traffic estimates, cordon counts, and vehicle speed data and supplemented by interviews with freight industry experts and operators. Findings indicate that freight traffic increased throughout London during this timeframe, but declined in the central boroughs partly within the congestion charge zone. The congestion charge may have time-shifted some light goods trips, but most freight trips face a variety of constraints on operators’ delivery window. No evidence was found of re-routing of freight traffic or avoidance traffic around the charged zone. The low emission zone spurred higher levels of operational change than the congestion charge zone, and it was effective at spurring freight vehicle replacement. The paper also discusses freight operators’ perceptions of these policies and how they could be improved.