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.

Intake of dairy products in relation to periodontitis in older Danish adults

This cross-sectional study investigates whether calcium intakes from dairy and non-dairy sources, and absolute intakes of various dairy products, are associated with periodontitis. The calcium intake (mg/day) of 135 older Danish adults was estimated by a diet history interview and divided into dairy and non-dairy calcium. Dairy food intake (g/day) was classified into four groups: milk, cheese, fermented foods and other foods. Periodontitis was defined as the number of teeth with attachment loss ≥3 mm. Intakes of total dairy calcium (Incidence-rate ratio (IRR) = 0.97; p = 0.021), calcium from milk (IRR = 0.97; p = 0.025) and fermented foods (IRR = 0.96; p = 0.03) were inversely and significantly associated with periodontitis after adjustment for age, gender, education, sucrose intake, alcohol consumption, smoking, physical activity, vitamin D intake, heart disease, visits to the dentist, use of dental floss and bleeding on probing, but non-dairy calcium, calcium from cheese and other types of dairy food intakes were not. Total dairy foods (IRR = 0.96; p = 0.003), milk (IRR = 0.96; p = 0.028) and fermented foods intakes (IRR = 0.97; p = 0.029) were associated with reduced risk of periodontitis, but cheese and other dairy foods intakes were not. These results suggest that dairy calcium, particularly from milk and fermented products, may protect against periodontitis. Prospective studies are required to confirm these findings.