Improved biocompatibility of protein encapsulation in sol-gel materials

By using the fluorescent dye 6-propionyl-2-(N,N-dimethylamino) naphthalene (PRODAN) to monitor methanol generated during tetramethyl orthosilicate polymerization we have optimised the encapsulation of protein in silica sol-gel monoliths with respect to completion of hydrolysis and distillation in order to remove methanol such that protein can be added without denaturation. A minimum of 24 h at +4 °C was found to be required before hydrolysis is complete and 3-5 min of vacuum distillation at 50 °C and 300 mbar needed to remove methanol before the gel is formed. The biocompatibility of a tetramethyl orthosilicate sol-gel monolith was demonstrated by preserving the trimer protein allophycocyanin (APC) in its native form for up to 500 h. This obviates the previously essential requirement of covalently binding the trimer together in order to prevent dissociation into monomers and has enabled observation of native APC trimer in a sol-gel pore for the first time down to the single molecule level using combined fluorescence spectroscopy and confocal microscopy. The higher stability afforded by the protocol we describe could impact on the application of sol-gel materials to single-molecule studies of wider bearing such as protein folding and aggregation

Inter-specific synchrony of two contrasting ungulates: wild boar (Sus scrofa) and roe deer (Capreolus capreolus)

Very few studies on ungulates address issues of inter-specific synchrony in population responses to environmental variation such as climate. Depending on whether annual variation in performance of ungulate populations is driven by direct or indirect (trophic) interactions, very different predictions regarding the pattern of inter-specific synchrony can be derived. We compared annual autumn body mass variation in roe deer (Capreolus capreolus) and wild boar (Sus scrofa) from Poland over the period 1982–2002, and related this to variation in winter and summer climate and plant phenological development [the Normalized Difference Vegetation Index (NDVI), derived from satellites]. Roe deer fawns (∼1.3 kg increase from year 1982 to 2002) and yearlings both increased markedly in mass over years. There was also an increase for wild boar mass over years (∼4.2 kg increase for piglets from 1982 to 2002). Despite our failure to link annual body mass to spring or winter conditions or the NDVI, the body mass of roe deer and wild boar fluctuated in synchrony. As this was a field roe deer population, and since wild boar is an omnivore, we suggest this may be linked to annual variation and trends in crop structure (mainly rye). We urge future studies to take advantage of studying multiple species in order to gain further insight into processes of how climate affect ungulate populations

Dynamics of major histocompatibility complex class II-positive cells in the postischemic brain - influence of levodopa treatment

Background: Cerebral ischemia activates both the innate and the adaptive immune response, the latter being activated within days after the stroke onset and triggered by the recognition of foreign antigens. Methods: In this study we have investigated the phenotype of antigen presenting cells and the levels of associated major histocompatibility complex class II (MHC II) molecules in the postischemic brain after transient occlusion of the middle cerebral artery (tMCAO) followed by levodopa/benserazide treatment. Male Sprague Dawley rats were subjected to tMCAO for 105 minutes and received levodopa (20 mg/kg)/benserazide (15 mg/kg) for 5 days starting on day 2 after tMCAO. Thereafter, immune cells were isolated from the ischemic and contralateral hemisphere and analyzed by flow cytometry. Complementarily, the spatiotemporal profile of MHC II-positive (MHC II+) cells was studied in the ischemic brain during the first 30 days after tMCAO; protein levels of MHC II and the levels of inflammation associated cytokines were determined in the ischemic hemisphere. Results: We found that microglia/macrophages represent the main MHC II expressing cell in the postischemic brain one week after tMCAO. No differences in absolute cell numbers were found between levodopa/benserazide and vehicle-treated animals. In contrast, MHC II protein levels were significant downregulated in the ischemic infarct core by levodopa/benserazide treatment. This reduction was accompanied by reduced levels of IFN-gamma, TNF-alpha and IL-4 in the ischemic hemisphere. In the contralateral hemisphere, we exclusively detected MHC II+ cells in the corpus callosum. Interestingly, the number of cells was increased by treatment with levodopa/benserazide independent from the infarct size 14 days after tMCAO. Conclusions: Results suggest that dopamine signaling is involved in the adaptive immune response after stroke and involves microglia/macrophages