A Novel Mechanism for Antibody-based Anthrax Toxin Neutralization: INHIBITION OF PREPORE-TO-PORE CONVERSION

Protective antigen (PA), a key component of anthrax toxin, mediates the entry of lethal factor (LF) or edema factor (EF) through a membranal pore into target cells. We have previously reported the isolation and chimerization of cAb29, an anti-PA monoclonal antibody that effectively neutralizes anthrax toxin in an unknown mechanism. The aim of this study was to elucidate the neutralizing mechanism of this antibody in vitro and to test its ability to confer post-exposure protection against anthrax in vivo. By systematic evaluation of the steps taking place during the PA-based intoxication process, we found that cAb29 did not interfere with the initial steps of intoxication, namely its ability to bind to the anthrax receptor, the consecutive proteolytic cleavage to PA₆₃, oligomerization, prepore formation, or LF binding. However, the binding of cAb29 to the prepore prevented its pH-triggered transition to the transmembranal pore, thus preventing the last step of intoxication, i.e. the translocation of LF/EF into the cell. Epitope mapping, using a phage display peptide library, revealed that cAb29 binds the 2α₁ loop in domain 2 of PA, a loop that undergoes major conformational changes during pore formation. In vivo, we found that 100% of anthrax-infected rabbits survived when treated with cAb29 12 h after exposure. In conclusion, these experiments demonstrate that cAb29 exerts its potent neutralizing activity in a unique manner by blocking the prepore-to-pore conversion process.     

Tropical Andean Forests Are Highly Susceptible to Nutrient Inputs—Rapid Effects of Experimental N and P Addition to an Ecuadorian Montane Forest

Tropical regions are facing increasing atmospheric inputs of nutrients, which will have unknown consequences for the structure and functioning of these systems. Here, we show that Neotropical montane rainforests respond rapidly to moderate additions of N (50 kg ha⁻¹ yr⁻¹) and P (10 kg ha⁻¹ yr⁻¹). Monitoring of nutrient fluxes demonstrated that the majority of added nutrients remained in the system, in either soil or vegetation. N and P additions led to not only an increase in foliar N and P concentrations, but also altered soil microbial biomass, standing fine root biomass, stem growth, and litterfall. The different effects suggest that trees are primarily limited by P, whereas some processes—notably aboveground productivity—are limited by both N and P. Highly variable and partly contrasting responses of different tree species suggest marked changes in species composition and diversity of these forests by nutrient inputs in the long term. The unexpectedly fast response of the ecosystem to moderate nutrient additions suggests high vulnerability of tropical montane forests to the expected increase in nutrient inputs.     

Advancing marine conservation planning in the Mediterranean Sea

Twenty leading scientists in the field of marine conservation planning attended the first international workshop on conservation planning in the Mediterranean Sea. This globally significant biodiversity hotspot has been subjected to human exploitation and degradation for 1,000s of years. Recently, several initiatives have tried to identify priority areas for conservation across the Mediterranean Sea. However, none of these efforts have led to large-scale actions yet. The aim of the workshop was to establish a network of scientists who are involved in large-scale conservation planning initiatives throughout the Mediterranean basin to promote collaboration and reduce redundancy in conservation initiatives. The three focus groups of the workshop build on existing efforts and intend to deliver: (1) a roadmap for setting conservation priorities, (2) a methodological framework for linking threats, actions and costs to improve the prioritization process, and (3) a systematic conservation planning process tailored to complex environments such as the Mediterranean Sea. Joining forces and involving more scientists (especially from the South-eastern part of the region) in following meetings, the participants endeavour to provide guidelines on how to bridge the science-policy gap and hence aid decision-makers to take efficient conservation actions.     

Inactivation of thiol-dependent enzymes by hypothiocyanous acid: role of sulfenyl thiocyanate and sulfenic acid intermediates

Myeloperoxidase (MPO) forms reactive oxidants including hypochlorous and hypothiocyanous acids (HOCl and HOSCN) under inflammatory conditions. HOCl causes extensive tissue damage and plays a role in the progression of many inflammatory-based diseases. Although HOSCN is a major MPO oxidant, particularly in smokers, who have elevated plasma thiocyanate, the role of this oxidant in disease is poorly characterized. HOSCN induces cellular damage by targeting thiols. However, the specific targets and mechanisms involved in this process are not well defined. We show that exposure of macrophages to HOSCN results in the inactivation of intracellular enzymes, including creatine kinase (CK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In each case, the active-site thiol residue is particularly sensitive to oxidation, with evidence for reversible inactivation and the formation of sulfenyl thiocyanate and sulfenic acid intermediates, on treatment with HOSCN (less than fivefold molar excess). Experiments with DAz-2, a cell-permeable chemical trap for sulfenic acids, demonstrate that these intermediates are formed on many cellular proteins, including GAPDH and CK, in macrophages exposed to HOSCN. This is the first direct evidence for the formation of protein sulfenic acids in HOSCN-treated cells and highlights the potential of this oxidant to perturb redox signaling processes.     

Direct recruitment of polycomb repressive complex 1 to chromatin by core binding transcription factors

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Highlights? PRC1 interacts with Runx1 and CBFβ ? PRC1 and Runx1/CBFβ colocalize on chromatin and coregulate a subset of genes ? PRC1 and Runx1/CBFβ deficiencies generate partial phenocopies of one another in vivo ? Runx1 directly recruits PRC1-independent of PRC2     

Tailored Enrichment Strategies and Stereotypic Behavior in Captive Individually Housed Macaques (Macaca spp.)

The welfare of nonhuman animals in captivity is widely dependent on the natural psychological, physical, and behavioral needs of the animals and how adequately these needs are met. Inability to engage in natural behaviors can lead to chronic stress and expression of stereotypic behavior. The majority of research on decreasing stereotypic behavior in captivity addresses problems at the group level and does not account for individual variability in each animal's needs, history, and preferences. This study combined physiological and behavioral measures of well being to comprehensively assess the unique needs of individually housed captive macaques (Macaca spp.) with the aim of developing tailored welfare strategies. Behavioral and hormonal data were collected under 2 conditions: baseline and individualized enrichment. The results showed a significant decrease in stereotypic behavior under the enrichment condition. Additionally, 7 out of 9 individuals showed a decrease in fecal glucocorticoid (stress hormone) levels, indicating a reduction in hypothalamic-pituitary-adrenal activity. Addressing welfare on an individual, rather than group, level allows for a better overall characterization of well being and maximizes the probability of improving the welfare of each animal.     

HIF-2α Expression Regulates Sprout Formation into 3D Fibrin Matrices in Prolonged Hypoxia in Human Microvascular Endothelial Cells

BackgroundDuring short-term hypoxia, Hypoxia Inducible Factors (particular their subunits HIF-1α and HIF-2α) regulate the expression of many genes including the potent angiogenesis stimulator VEGF. However, in some pathological conditions chronic hypoxia occurs and is accompanied by reduced angiogenesis.ObjectivesWe investigated the effect of prolonged hypoxia on the proliferation and sprouting ability of human microvascular endothelial cells and the involvement of the HIFs and Dll4/Notch signaling.ConclusionProlonged culturing of hMVECs at 1% oxygen inhibited endothelial sprouting into fibrin. Two independent mechanisms contribute. Silencing of HIF-2α with si-RNA partially restored the inhibition of endothelial sprouting pointing to a HIF-2α-dependent mechanism. In addition, reduction of uPA contributed to reduced endothelial tube formation in a fibrin matrix during prolonged hypoxia.     

The effects of previous experience and current condition on status contests in the bluebanded goby (<Emphasis Type="Italic">Lythrypnus dalli</Emphasis>)

Social status can change over a lifetime and affect fitness. Status transitions are often influenced by previous experience such that previous winners tend to dominate in future interactions, while losers tend to remain subordinate. We investigated the role of social status experience on future status contests in the bluebanded goby (Lythrypnus dalli), a highly social, sex-changing fish. Female L. dalli were given long-term experience as alpha (dominant) or beta (subordinate) in a stable social group of three fish (two females, one dominant male). We hypothesized that females with previous dominance experience would be more likely to become dominant in a novel context and change sex, a status-dependent transition. We found that in size-matched pairs, half of the previous alphas and half of the previous betas established and maintained dominance in the novel pair. Previous betas were more likely to become dominant in pairs with small relative differences in physical condition. Betas that initiated more interactions with the male in the stable group were also more likely to become dominant. Experience only had a transient effect on agonistic behavior during status establishment. In pairs in which the previous beta established dominance, individual agonistic efficiency, or the proportion of their approaches that led to a displacement, was lower. There was no long-term effect of experience on the rate of sex change. These data suggest that long-term tenure as a subordinate does not, itself, prevent status ascension and that physical condition and individual behavior are relevant factors that influence L. dalli contests.