Contribution of individual histidines to the global stability of human prolactin

A member of the family of hematopoietic cytokines human prolactin (hPRL) is a 23k kDa polypeptide hormone, which displays pH dependence in its structural and functional properties. The binding affinity of hPRL for the extracellular domain of its receptor decreases 500‐fold over the relatively narrow, physiologic pH range from 8 to 6; whereas, the affinity of human growth hormone (hGH), its closest evolutionary cousin, does not. Similarly, the structural stability of hPRL decreases from 7.6 to 5.6 kcal/mol from pH 8 to 6, respectively, whereas the stability of hGH is slightly increased over this same pH range. hPRL contains nine histidines, compared with hGH's three, and they are likely responsible for hPRL's pH‐dependent behavior. We have systematically mutated each of hPRL's histidines to alanine and measured the effect on pH‐dependent global stability. Surprisingly, a vast majority of these mutations stabilize the native protein, by as much as 2–3 kcal/mol. Changes in the overall pH dependence to hPRL global stability can be rationalized according to the predominant structural interactions of individual histidines in the hPRL tertiary structure. Using double mutant cycles, we detect large interaction free energies within a cluster of nearby histidines, which are both stabilizing and destabilizing to the native state. Finally, by comparing the structural locations of hPRL's nine histidines with their homologous residues in hGH, we speculate on the evolutionary role of replacing structurally stabilizing residues with histidine to introduce pH dependence to cytokine function.     

Is artificial habitat diversity a key to restoring nurseries for juvenile coastal fish? Ex situ experiments on habitat selection and survival of juvenile seabreams

Man‐made infrastructures have become ubiquitous components of coastal landscapes, leading to habitat modification that affects the abundance and diversity of marine organisms. Marine coastal fish have a complex life cycle requiring different essential habitats. One of these habitats is known as a nursery, a place where juveniles can settle in large numbers, survive, and grow to contribute to the adult population. Nurseries are mainly found in shallow, sheltered zones and are thus particularly impacted by urbanization, notably by harbors. The vertical featureless structure of docks is very unlikely to be used by juveniles, which need complex habitats to find food and shelter from predators. Recent attempts to rehabilitate the nursery function in such environments by using artificial habitats have proven efficient in increasing juvenile densities. However, nothing is known about the survival of juveniles in these habitats, preventing any conclusions on the effectiveness of this means of restoration from being drawn. Here, we set up tank experiments to test the relationship between habitat preferences and the survival rate of two species of seabream when facing stalk‐attacking combers. Habitat choice was consistent with survival results, indicating that artificial habitats might not represent unintended ecological traps for juveniles. However, the artificial habitats' effect on survival was variable between species. Therefore, our results suggest that habitat diversity might be of prime importance to sustain juveniles of different species and stress the need for the development of diverse artificial habitats to counteract the effects of seascape homogenization.     

Plant–parasite coevolution: A weak signature of local adaptation between Peruvian Globodera pallida populations and wild potatoes

Plant–parasite coevolution has generated much interest and studies to understand and manage diseases in agriculture. Such a reciprocal evolutionary process could lead to a pattern of local adaptation between plants and parasites. Based on the phylogeography of each partner, the present study tested the hypothesis of local adaptation between the potato cyst nematode Globodera pallida and wild potatoes in Peru. The measured fitness trait was the hatching of cysts which is induced by host root exudates. Using a cross‐hatching assay between 13 populations of G. pallida and root exudates from 12 wild potatoes, our results did not show a strong pattern of local adaptation of the parasite but the sympatric combinations induced better hatching of cysts than allopatric combinations, and there was a negative relationship between the hatching percentage and the geographical distance between nematode populations and wild potatoes. Moreover, a strong effect of the geographic origin of root exudates was found, with root exudates from south of Peru inducing better hatching than root exudates from north of Peru. These results could be useful to develop new biocontrol products or potato cultivars to limit damages caused by G. pallida.     

Neutron Imaging of Meat during Cooking

Neutron imaging was used to follow the impact of cooking on beef meat. During online cooking, the cartography realised on image collected shows neutron attenuation per zone. Some data points were taken on the edge to highlight higher attenuation variations because of “microscopic” shrinkage of the meat at 70 °C. Some others points were taken in the centre of the sample, which first showed smaller decreases at 75 °C and then an increase around 80 °C. These smaller attenuation variations are possibly linked to denaturation of connective tissue which in turn influenced meat microstructure allowing the release of entrapped water and increase the thickness of the sample.     

CD40 Gene Silencing Reduces the Progression of Experimental Lupus Nephritis Modulating Local Milieu and Systemic Mechanisms

Lupus nephritis (LN) is an autoimmune disorder in which co-stimulatory signals have been involved. Here we tested a cholesterol-conjugated-anti-CD40-siRNA in dendritic cells (DC) in vitro and in a model of LPS to check its potency and tissue distribution. Then, we report the effects of Chol-siRNA in an experimental model of mice with established lupus nephritis. Our in vitro studies in DC show a 100%intracellular delivery of Chol-siRNA, with a significant reduction in CD40 after LPS stimuli. In vivo in ICR mice, the CD40-mRNA suppressive effects of our Chol-siRNA on renal tissue were remarkably sustained over a 5 days after a single preliminary dose of Chol-siRNA. The intra-peritoneal administration of Chol-siRNA to NZB/WF1 mice resulted in a reduction of anti-DNA antibody titers, and histopathological renal scores as compared to untreated animals. The higher dose of Chol-siRNA prevented the progression of proteinuria as effectively as cyclophosphamide, whereas the lower dose was as effective as CTLA4. Chol-siRNA markedly reduced insterstitialCD3+ and plasma cell infiltrates as well as glomerular deposits of IgG and C3. Circulating soluble CD40 and activated splenic lymphocyte subsets were also strikingly reduced by Chol-siRNA. Our data show the potency of our compound for the therapeutic use of anti-CD40-siRNA in human LN and other autoimmune disorders.     

Developmental compartmentalization in the dorsal mesothoracic disc of Drosophila.

The clonal analysis of the development of the dorsal mesothoracic (wing) disc shows that clones initiated after a given time do not cross over certain demarcation lines in the adult cuticle. The property of $\text{M}{}^{\mathrm{+}}\text{M}{}^{\mathrm{+}}$ (non-Minute) recombinant cells to overgrow the $\text{M}\text{M}{}^{\mathrm{+}}$ background cells was used to demonstrate the establishment of clonal restrictions during development. It has been shown that $\text{M}{}^{\mathrm{+}}\text{M}{}^{\mathrm{+}}$ clones initiated at a given time of development share common demarcation lines that delimit what we call “a developmental compartment.” As development time and cell proliferation of the anlage proceed, large compartments become split into pairs of smaller ones. A study of the number of cells in a given compartment at the time of its splitting into subcompartments indicates that the “developmental segregation” takes place in groups of neighboring cells and suggests that the number of segregated cells is different and characteristic for each compartment. Within a given compartment, a single clone of $\text{M}{}^{\mathrm{+}}\text{M}{}^{\mathrm{+}}$ cells gives rise to 60–90% of the total number of adult cells. This phenomenon is reminiscent of the regulative properties of the morphogenetic fields, and the relation of these to developmental compartments is discussed. Since homoeotic mutants transform entire developmental compartments into one another, the hypothesis is advanced that homoeotic genes control compartment development.