Complete structure of an increasing capillary permeability protein (ICPP) purified from Vipera lebetina venom. ICPP is angiogenic via vascular endothelial growth factor receptor signalling

The partial sequence of the increasing capillary permeability protein (ICPP) purified from Vipera lebetina venom revealed a strong homology to vascular endothelial growth factor (VEGF)-A. We now report its complete amino acid sequence determined by Edman degradation and its biological effects on mouse and human vascular endothelial cells. ICPP is a homodimeric protein linked by cysteine disulfide bonds of 25115 Da revealed by mass spectrometry. Each monomer is composed of 110 amino acids including eight cysteine residues and a pyroglutamic acid at the N-terminal extremity. ICPP shares 52% sequence identity with human VEGF but lacks the heparin binding domain and Asn glycosylation site. Besides its strong capillary permeability activity, ICPP was found to be a potent in vitro angiogenic factor when added to mouse embryonic stem cells or human umbilical vein endothelial cells. ICPP was found to be as potent as human VEGF165 in activating p42/p44 MAPK, in reinitiation of DNA synthesis in human umbilical vein endothelial cells, and in promoting in vitro angiogenesis of mouse embryonic stem cells. All these biological actions, including capillary permeability in mice, were fully inhibited by 1 microm of a new specific VEGF receptor tyrosine kinase inhibitor (ZM317450) from AstraZeneca that belongs to the anilinocinnoline family of compounds. Indeed, up to a 30 times higher concentration of inhibitor did not affect platelet-derived growth factor, epidermal growth factor, FGF-2, insulin, alpha-thrombin, or fetal calf serum-induced p42/p44 MAPK and reinitiation of DNA synthesis. Therefore, we conclude that this venom-derived ICPP exerts its biological action (permeability and angiogenesis) through activation of VEGF receptor signaling (VEGF-R2 and possibly VEGF-R1).     

Disturbance-dependent Yellow-legged Gull (Larus michahellis) predation on Larid chicks decreases with chick age

Predation is one of the key factors shaping the dynamics of animal populations. In birds, nest loss due to predation can be a significant cause of low reproductive success. Ground-nesting birds are among the bird groups most susceptible to predation, mainly because their nests are easily accessible to a broad suite of potential predators. For these birds, anthropogenic disturbances can generate changes in nest predation risk by altering their antipredator behaviour and also by altering the behaviour of the predator species, i.e. the predator becoming much more aware of predation opportunities due to frequent disturbances and/or motivated to repeat predation attempts when some are successful. To date, most previous studies investigating this have focused on a single effect, either predation or disturbance, on chick survival. It remains unknown how the risk of predation with and without disturbance varies with chick age. In this study, we used behavioural observations to assess how the interaction between predators and disturbance affects predation risk in chicks and how this interacts with chick age. Specifically, we investigated the effect of disturbance caused by humans and stray dogs on the predation of Slender-billed Gull Chroicocephalus genei chicks by Yellow-legged Gulls Larus michahellis, and whether this depended on the age of the chicks. Our results revealed that disturbance had a significant positive effect on predation measures of Slender-billed Gull chicks by Yellow-legged Gulls, but that this effect was mediated both by disturbance type and the age of chicks. Stray dogs entering the colony had a stronger disturbance effect on chicks than passing humans, increasing predation risk by Yellow-legged Gulls. Our results also showed that chick age interacts with disturbance type to determine the predation risk. This is probably mediated by chicks' capacity to escape predation by gathering in a single large crèche that runs into the water when disturbed. To preserve Slender-billed Gull colonies in one of its few remaining breeding sites in Tunisia, and as gulls tend to react even when the disturbance occurs relatively far from the colonies, it is crucial to (1) restrict human access to dikes and islets where large colonies breed and (2) construct artificial islets attractive to gulls and inaccessible to stray dogs.     

Identification of calmodulin-binding proteins in brain of worker honeybees

Calmodulin is a Ca(+2)-binding protein important in a variety of cell functions. The Ca(+2)/calmodulin complex interacts with and regulates various enzymes and target proteins, known as calmodulin-binding proteins (CaMBPs). In this study, we revealed a comparative identification of the CaMBPs composition in the worker honeybee (Apis mellifera) brain, considering two different honeybee behaviors in the colony. To this end, the CaMBPs of forager and nurse workers were purified by affinity chromatography, separated in 1D gel, digested and submitted to peptide mass fingerprinting (PMF). In the PMF analysis, 15 different proteins, considered behavior-specific proteins, were identified, one of them exclusively in forager workers and 10 in nurses. All the proteins were classified in terms of their function and cell localization, revealing a greater expression of metabolism-related CaMBPs in both worker subcastes. Protein sequences were then analyzed for the presence of the calmodulin-binding sites. Therefore, the honeybee brain CaMBPs profiles presented differences between worker subcastes. This is the first identification of calmodulin-binding proteins in the brain of A. mellifera upon nursing and foraging behaviors in the colony and this diversity of target proteins for Ca(+2)/CaM may be involved in terms of the function of these proteins in the nervous system.     

Diversity of waterbirds wintering in Douz wetlands (south Tunisia): factors affecting wetland occupancy and species richness

Understanding the environmental factors shaping wetland attractiveness for waterbirds is an important question in wetland ecology and for conservation purposes. However, detailed data from numerous North African wetlands, notably those situated in inland areas, are still lacking. Thus, the aim of this study was to identify the factors influencing wetland use by waterbirds wintering in one of such poorly known habitat systems, namely the Saharan wetland complex of Douz, in south-western Tunisia. Thirty-four waterbird species (20 wading birds and 14 waterfowl species) were found to winter in this area. Using discriminant function analyses, we found that wetland size was the unique variable discriminating between occupied and unoccupied sites for total waterbirds and wading birds, while waterfowl distribution was related to both wetland size and water depth. Wetland size also provided the strongest predictor of species richness of wading birds, waterfowl and total waterbirds. Overall, our findings highlight the importance of wetland size as a key factor determining the attractiveness of wetlands for waterbirds wintering in the Saharan wetland complex of Douz. The possible explanations of this wetland size effect are discussed.     

Novel svVEGF isoforms from Macrovipera lebetina venom interact with neuropilins

Increased vascular permeability and vasodilation are responses usually elicited by snake envenomation. In this report, we isolated from Macroviperalebetina venom two protein groups designated IC1 (Increasing Capillary1) and IC2 based on their activities on capillary permeability. Mass spectrometry analysis showed that IC1 contained four major proteins of 23,650, 24,306, 24,589 and 24,718 Da, whereas IC2 contained three major proteins of 25,101, 25,194 and 25,298 Da. N-terminal amino-acid sequencing revealed that IC1 and IC2 belong to the snake venom VEGF (svVEGF) family. IC1 and IC2 had a marked specificity for VEGFR-2, with affinities in the nanomolar range. Interestingly, they also bind to NRP1 and NRP2, with affinities in the micromolar range. This is the first report demonstrating that M. lebetina encodes several distinct svVEGFs, endowed with a capacity to interact with neuropilins. IC1 and IC2 could be valuable tools to understand the molecular properties of angiogenic factors and their receptors.     

On the mechanisms of cadmium stress alleviation in Medicago truncatula by arbuscular mycorrhizal symbiosis: A root proteomic study

The arbuscular mycorrhizal (AM) symbiosis belongs to the strategies plants have developed to cope with adverse environmental conditions including contamination by heavy metals such as cadmium (Cd). In the present work, we report on the protective effect conferred by AM symbiosis to the model legume Medicago truncatula grown in presence of Cd, and on the 2‐D‐based proteomic approach further used to compare the proteomes of M. truncatula roots either colonised or not with the AM fungus Glomus intraradices in Cd‐free and Cd‐contaminated substrates. The results indicated that at the proteome level, 9 out of the 15 cadmium‐induced changes in nonmycorrhizal roots were absent or inverse in those Cd‐treated and colonized by G. intraradices, including the G. intraradices‐dependent down‐accumulation of Cd stress‐responsive proteins. Out of the twenty‐six mycorrhiza‐related proteins that were identified, only six displayed changes in abundance upon Cd exposure, suggesting that part of the symbiotic program, which displays low sensitivity to Cd, may be recruited to counteract Cd toxicity through the mycorrhiza‐dependent synthesis of proteins having functions putatively involved in alleviating oxidative damages, including a cyclophilin, a guanine nucleotide‐binding protein, an ubiquitin carboxyl‐terminal hydrolase, a thiazole biosynthetic enzyme, an annexin, a glutathione S‐transferase (GST)‐like protein, and a S‐adenosylmethionine (SAM) synthase.     

miR-216 and miR-217 expression is reduced in transgenic mouse models of pancreatic adenocarcinoma,knockout of miR-216/miR-217 host gene is embryonic lethal

Mice harboring a G12D activating Kras mutation are among the most heavily studied models in the field of pancreatic adenocarcinoma (PDAC) research. miRNAs are differentially expressed in PDAC from patients and mouse models of PDAC. To better understand the relationship that Kras activation has on miRNA expression, we profiled the expression of 629 miRNAs in RNA isolated from the pancreas of control, young, and old P48^+/Cre;LSL-KRAS^G12D as well as PDX-1-Cre;LSL-KRAS^G12D mice. One hundred of the differentially expressed miRNAs had increased expression in the advanced disease (old) P48^+/Cre;LSL-KRAS^G12D compared to wild-type mice. Interestingly, the expression of three miRNAs, miR-216a, miR-216b, and miR-217, located within a ～30-kbp region on 11qA3.3, decreased with age (and phenotype severity) in these mice. miR-216/-217 expression was also evaluated in another acinar-specific ELa-Kras^G12D mouse model and was downregulated as well. As miR-216/-217 are acinar enriched, reduced in human PDAC and target KRAS, we hypothesized that they may maintain acinar differentiation or represent tumor suppressive miRNAs. To test this hypothesis, we deleted a 27.9-kbp region of 11qA3.3 containing the miR-216/-217 host gene in the mouse’s germ line. We report that germ line deletion of this cluster is embryonic lethal in the mouse. We estimate that lethality occurs shortly after E9.5. qPCR analysis of the miR-216b and miR-217 expression in the heterozygous animals showed no difference in expression, suggesting haplosufficiency by some type of compensatory mechanism. We present the differential miRNA expression in Kras^G12D transgenic mice and report lethality from deletion of the miR-216/-217 host gene in the mouse’s germ line.     

Assortative mating for carotenoid colouration but not size in the Yellow-legged Gull Larus michahellis

Capsule: Mediterranean Yellow-legged Gulls mate assortatively according to carotenoid-based colouration but not in relation to size.