Evolution of Afrotropical freshwater crab lineages obscured by morphological convergence

We use sequence data derived from six DNA gene loci to examine evolutionary and biogeographic affinities among all freshwater crab families. With an emphasis on the Afrotropical fauna that includes Africa, Madagascar, and the Seychelles, we test the proposed Gondwanan cladogenesis of the group. Phylogenetic results demonstrate that contemporary distribution patterns of freshwater crab lineages are incongruent with the expected area cladogram of continental fragmentation. Instead, our phylogenetic estimate and divergence time estimation indicate a post-Gondwanan, early Cretaceous cladogenesis for freshwater crabs implying that the acquisition of a freshwater lifestyle was achieved more recently. A dispersal hypothesis as opposed to vicariance appears to best explain the contemporary distribution pattern of this group. However, our results do not explicitly disprove a Gondwanan origin for the Afrotropical freshwater crabs. Alarmingly, these results suggest that most of the currently recognized freshwater crab families are unreliable taxonomic groupings since virtually no Afrotropical freshwater crab families formed monophyletic units thus obscuring inferred biogeographic relationships. Convergence in characters associated with the terminal segment of the mandibular palp is clearly a pervasive obstacle in the taxonomy of this group.     

Renal IL-17 expression in human ANCA-associated glomerulonephritis

Interleukin-17A (IL-17) promotes inflammatory renal tissue damage in mouse models of crescentic glomerulonephritis, including murine experimental autoimmune anti-myeloperoxidase glomerulonephritis, which most likely depends on IL-17-producing Th17 cells. In human anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis, however, the cellular sources of IL-17 remain to be elucidated. Therefore, we analyzed human kidney biopsies of active necrotizing and crescentic ANCA-associated glomerulonephritis by immunohistochemistry using an IL-17-specific antibody and by immunofluorescent colocalization with cell type markers. We detected numerous IL-17-expressing (IL-17(+)) cells in the glomeruli and in the tubulointerstitium. Unexpectedly, most of these IL-17(+) cells were polymorphonuclear neutrophilic granulocytes, while IL-17(+) T cells and IL-17(+) mast cells were present at significantly lower frequencies. IL-17 was not detected in other infiltrating or resident kidney cells. In those patients who had not received immunosuppressive treatment before biopsy, serum creatinine levels were positively correlated with tubulointerstitial IL-17(+) neutrophils as well as IL-17(+) T cells. Furthermore, we could demonstrate that purified human blood neutrophils expressed IL-17 protein and released it upon stimulation in vitro. In conclusion, these results support a pathogenic role for IL-17 in human ANCA-associated glomerulonephritis. Our data suggest that in the acute stage of the disease neutrophils may act as an important immediate-early innate source of IL-17 and may thereby initiate and promote ongoing renal inflammation. IL-17 may thus be a target for treating acute ANCA-associated glomerulonephritis.     

Influence of drying on the secondary structure of intrinsically disordered and globular proteins

Circular dichroism (CD) spectroscopy of five Arabidopsis late embryogenesis abundant (LEA) proteins constituting the plant specific families LEA_5 and LEA_6 showed that they are intrinsically disordered in solution and partially fold during drying. Structural predictions were comparable to these results for hydrated LEA_6, but not for LEA_5 proteins. FTIR spectroscopy showed that verbascose, but not sucrose, strongly affected the structure of the dry proteins. The four investigated globular proteins were only mildly affected by drying in the absence, but strongly in the presence of sugars. These data highlight the larger structural flexibility of disordered compared to globular proteins and the impact of sugars on the structure of both disordered and globular proteins during drying.     

The effect of species diversity on lipid production by micro-algal communities

Current research investigating the importance of diversity for biofuel lipid production remains limited. In contrast, the relationship between diversity and productivity within terrestrial and algal primary producers has been well documented in ecology. Hence, we set out to investigate, experimentally, whether diversity may also affect lipid production in micro-algae. We investigated the growth and lipid production of micro-algae using species from all major algal groups. Algae were grown in a large number of treatments differing in their diversity level. Additionally, we compared the growth and lipid production of laboratory communities to natural lake and pond phytoplankton communities of different diversity. Our results show that lipid production increased with increasing diversity in both natural and laboratory micro-algal communities. The underlying reason for the observed ‘diversity–productivity’ relationship seems to be resource use complementarity. We observed higher lipid production of highly diverse algal communities under the same growth and resource supply conditions compared to monocultures. Hence, the incorporation of the ecological advantages of diversity-related resource-use dynamics into algal biomass production may provide a powerful and cost effective way to improve biofuel production.     

Direct fluorimetric sensing of UV-excited analytes in biological and environmental samples using molecularly imprinted polymer nanoparticles and fluorescence polarization

A rapid, robust, sensitive and economic sensing method, based on a molecularly imprinted polymer (MIP) synthetic antibody mimic, and fluorescence polarization analysis, for the direct detection of UV-excited fluorescent analytes in food and environmental samples was developed. Fluoroquinolone (FQ) antibiotics were used as fluorescent model analytes. Water-compatible MIP nanoparticles were synthesized with enrofloxacin (ENRO) as the imprinting template. Fluorescence polarization measurements then allow the direct determination of the amount of ENRO and other structurally related piperazine-based fluoroquinolones that bind to the MIP. No separation step was required since this technique distinguishes in situ analyte molecules bound to the MIP from the free analyte in solution. This assay was successfully applied for the first time to determine FQs in real samples, i.e. tap water and milk, without any prior concentration step, by simply adding a known amount of MIP. No interference by the sample components was observed even though the excitation was in the UV region. In tap water, a low limit of detection of 0.1 nM for ENRO was achieved with 5 μg mL(-1) of MIP. In milk, ENRO and danofloxacin, whose MRLs have been fixed at 0.28 μM and 0.08 μM, respectively, could be selectively measured and distinguished from other families of antibiotics. The procedure is very easy and practical as it consists of simply precipitating the milk proteins with acetonitrile and adding buffer and MIP to the supernatant before reading the polarization values with a spectrofluorimeter.