Conservation of Freshwater Biodiversity: a Comparison of Different Area Selection Methods

The biodiversity of freshwater systems is endangered, especially in Mediterranean semiarid areas such as the south east of the Iberian Peninsula, whose rich and endemic biota is threatened by the development of surrounding land-crop irrigation. For this reason, the prioritization of areas for biodiversity conservation is an urgent target. In this study we used data records of water beetles from a province of the southeast of Spain for assessing priority areas for freshwater biodiversity conservation. We compare the performance of various area-selection methods, ranging from scoring procedures to complementarity-based algorithms, which are based on different criteria such as richness, rarity and vulnerability. The complementarity approaches were more efficient than methods using scoring or richness and rarity hotspots for representing conservation targets in a given number of areas and for identifying the minimum set of areas containing all species at least once. Within these, the richness-based algorithm was more efficient than rarity-based algorithm. Crucial target habitats for aquatic biodiversity conservation in the area studied are streams at medium altitude, hypersaline streams, and endorreic and karstic complexes.     

Successful colon cancer eradication after chemoimmunotherapy is associated with profound phenotypic change of intratumoral myeloid cells

IL-12 is a potent immunostimulatory cytokine, but its impact as an antitumor drug in clinical practice is limited. Upsurge of regulatory T cells (Treg) in the tumor milieu has been proposed to limit the efficacy of the treatment. In this paper, two drugs (cyclophosphamide [CPA] and anti-CD25 mAb) widely used to eliminate Treg were used in an attempt to enhance the antitumor effect of IL-12 gene therapy. Both anti-CD25 and CPA combined with IL-12 were able to deplete intratumoral Treg and myeloid-derived suppressor cells (MDSC), but only IL-12 plus CPA achieved significant antitumor activity in mice with large established s.c. colon carcinoma. This therapeutic effect was associated with the emergence of a heterogeneous population of myeloid cells within the tumor, termed inflammatory myeloid cells (IMC), composed of Ly6C(high)Ly6G(low) inflammatory monocytes and Ly6G(high)Ly6C(+) neutrophils. IMC showed a distinctive pattern of cytokine/chemokine production, and in contrast to MDSC, they did not induce conversion of naive CD4(+) T cells into Treg. The appearance of IMC coincided with intense tumor infiltration by effector T cells, which was abrogated by elimination of IMC by anti-Gr1 mAb, a maneuver that abolished the antitumor effect of the therapy. Therefore, the combination of IL-12 and CPA eliminates intratumoral Treg and MDSC, while it induces the appearance of IMC within the tumor microenvironment. The latter effect is essential to facilitate effector T cell infiltration and subsequent tumor elimination.     

Vascular smooth muscle relaxation by a lectin from Pisum arvense: evidences of endothelial NOS pathway

The vasorelaxant effect of the lectin of Pisum arvense (PAL) seeds was investigated in rat aorta. PAL (10-100 μg/ml) was applied on aorta rings, with or without endothelium, pre-contracted with phenylephrine (Phe; 0.1 μM). Participation of endothelium derived relaxant factors was evaluated incubating the tissue with indomethacin (10 μM), L-nitro arginine methyl ester (L-NAME, 100 μM) and tetraethylammonium (TEA, 5 mM) before addition of PAL. The role of the lectin domain was investigated by addition of PAL into tissue in presence of glucose (3x 10?? M), or N-acetyl Dglucosamine (GlcNAc; 3 x 10?? M). The importance of extracellular calcium (Ca2?e) or interaction with muscarinic receptors in the relaxant effect was evaluated by addition of PAL into aorta rings containing calcium free solution (OCa) and atropine (1 μ M), respectively. PAL induced concentration-dependent relaxation in endothelized aorta (IC50 =58.38 ± 1.87 μg/ml), which was reversed by L-NAME and glucose. The lectin effect was totally inhibited when the preparation was inserted in OCa, but not in presence of atropine. Summarizing, our data showed a relaxant effect of PAL in isolated rat aorta rings in presence of endothelium, suggestive of interaction between the lectin carbohydrate binding sites with specific receptors located in vascular endothelial cells leading to nitric oxide synthase activation. This effect seems to require Ca2?e but is independent on muscarinic receptors interaction.     

Development of functional symbiotic white clover root hairs and nodules requires tightly regulated production of rhizobial cellulase CelC2

The establishment of rhizobia as nitrogen-fixing endosymbionts within legume root nodules requires the disruption of the plant cell wall to breach the host barrier at strategic infection sites in the root hair tip and at points of bacterial release from infection threads (IT) within the root cortex. We previously found that Rhizobium leguminosarum bv. trifolii uses its chromosomally encoded CelC2 cellulase to erode the noncrystalline wall at the apex of root hairs, thereby creating the primary portal of its entry into white clover roots. Here, we show that a recombinant derivative of R. leguminosarum bv. trifolii ANU843 that constitutively overproduces the CelC2 enzyme has increased competitiveness in occupying aberrant nodule-like root structures on clover that are inefficient in nitrogen fixation. This aberrant symbiotic phenotype involves an extensive uncontrolled degradation of the host cell walls restricted to the expected infection sites at tips of deformed root hairs and significantly enlarged infection droplets at termini of wider IT within the nodule infection zone. Furthermore, signs of elevated plant host defense as indicated by reactive oxygen species production in root tissues were more evident during infection by the recombinant strain than its wild-type parent. Our data further support the role of the rhizobial CelC2 cell wall-degrading enzyme in primary infection, and show evidence of its importance in secondary symbiotic infection and tight regulation of its production to establish an effective nitrogen-fixing root nodule symbiosis.     

Relative Effects of Road Risk,Habitat Suitability,and Connectivity on Wildlife Roadkills: The Case of Tawny Owls (Strix aluco)

Background

Despite its importance for reducing wildlife-vehicle collisions, there is still incomplete understanding of factors responsible for high road mortality. In particular, few empirical studies examined the idea that spatial variation in roadkills is influenced by a complex interplay between road-related factors, and species-specific habitat quality and landscape connectivity.

Methodology/Principal Findings

In this study we addressed this issue, using a 7-year dataset of tawny owl (Strix aluco) roadkills recorded along 37 km of road in southern Portugal. We used a multi-species roadkill index as a surrogate of intrinsic road risk, and we used a Maxent distribution model to estimate habitat suitability. Landscape connectivity was estimated from least-cost paths between tawny owl territories, using habitat suitability as a resistance surface. We defined 10 alternative scenarios to compute connectivity, based on variation in potential movement patterns according to territory quality and dispersal distance thresholds. Hierarchical partitioning of a regression model indicated that independent variation in tawny owl roadkills was explained primarily by the roadkill index (70.5%) and, to a much lesser extent, by landscape connectivity (26.2%), while habitat suitability had minor effects (3.3%). Analysis of connectivity scenarios suggested that owl roadkills were primarily related to short range movements (<5 km) between high quality territories. Tawny owl roadkills were spatially autocorrelated, but the introduction of spatial filters in the regression model did not change the type and relative contribution of environmental variables.

Conclusions

Overall, results suggest that road-related factors may have a dominant influence on roadkill patterns, particularly in areas like ours where habitat quality and landscape connectivity are globally high for the study species. Nevertheless, the study supported the view that functional connectivity should be incorporated whenever possible in roadkill models, as it may greatly increase their power to predict the location of roadkill hotspots.     

ATR and Chk1 Suppress a Caspase-3–Dependent Apoptotic Response Following DNA Replication Stress

The related PIK-like kinases Ataxia-Telangiectasia Mutated (ATM) and ATM- and Rad3-related (ATR) play major roles in the regulation of cellular responses to DNA damage or replication stress. The pro-apoptotic role of ATM and p53 in response to ionizing radiation (IR) has been widely investigated. Much less is known about the control of apoptosis following DNA replication stress. Recent work indicates that Chk1, the downstream phosphorylation target of ATR, protects cells from apoptosis induced by DNA replication inhibitors as well as IR. The aim of the work reported here was to determine the roles of ATM- and ATR-protein kinase cascades in the control of apoptosis following replication stress and the relationship between Chk1-suppressed apoptotic pathways responding to replication stress or IR. ATM and ATR/Chk1 signalling pathways were manipulated using siRNA-mediated depletions or specific inhibitors in two tumour cell lines or fibroblasts derived from patients with inherited mutations. We show that depletion of ATM or its downstream phosphorylation targets, NBS1 and BID, has relatively little effect on apoptosis induced by DNA replication inhibitors, while ATR or Chk1 depletion strongly enhances cell death induced by such agents in all cells tested. Furthermore, early events occurring after the disruption of DNA replication (accumulation of RPA foci and RPA34 hyperphosphorylation) in ATR- or Chk1-depleted cells committed to apoptosis are not detected in ATM-depleted cells. Unlike the Chk1-suppressed pathway responding to IR, the replication stress-triggered apoptotic pathway did not require ATM and is characterized by activation of caspase 3 in both p53-proficient and -deficient cells. Taken together, our results show that the ATR-Chk1 signalling pathway plays a major role in the regulation of death in response to DNA replication stress and that the Chk1-suppressed pathway protecting cells from replication stress is clearly distinguishable from that protecting cells from IR.     

Microvascular pressure and functional capillary density in extreme hemodilution with low- and high-viscosity dextran and a low-viscosity Hb-based O2 carrier

Blood losses are usually corrected initially by the restitution of volume with plasma expanders and subsequently by the restoration of oxygen-carrying capacity using either a blood transfusion or possibly, in the near future, oxygen-carrying plasma expanders. The present study was carried out to test the hypothesis that high-plasma viscosity hemodilution maintains perfused functional capillary density (FCD) by preserving capillary pressure. Microvascular pressure responses to extreme hemodilution with low- (LV) and high-viscosity (HV) plasma expanders and an exchange transfusion with a polymerized bovine cell-free Hb (PBH) solution were analyzed in the awake hamster window chamber model (n = 26). Systemic hematocrit was reduced from 50% to 11%. PBH produced a greater mean arterial blood pressure than the nonoxygen carriers. FCD was higher after a HV plasma expander (70 +/- 15%) vs. PBH (47 +/- 12%). Microvascular pressure spanning the capillary network was higher after a HV plasma expander (16-19 mmHg) compared with PBH (12-16 mmHg) and a LV plasma expander (11-14 mmHg) but lower than control (22-26 mmHg). FCD was found to be directly proportional to capillary pressure. The use of a HV plasma expander in extreme hemodilution maintained the number of perfused capillaries and tissue perfusion by comparison with a LV plasma expander due to increased mean arterial blood pressure and capillary pressure. The use of PBH increased mean arterial pressure but reduced capillary pressure due to vasoconstriction and did not maintain FCD.     

Modelling enrolment in Cambrian trilobites

Trilobites were capable of enroling in different ways based on the flexible articulation of thoracic segments and associated interlocking devices; the type of enrolment (spiral or sphaeroidal) is thought to have largely depended on the coaptative devices that each trilobite used to enclose the body. Based on X‐ray microtomography scans of complete enrolled specimens from the Cambrian, we created three‐dimensional (3D) computer models to assess the kinematics needed to achieve both enrolment types. We demonstrate that closely related trilobites with little morphological variation (Bailiaspis?, Conocoryphe and Parabailiella) developed different enrolment types as a result of small variations in the number of thoracic segments and the angle between adjacent segments. Moreover, our models indicate that sphaeroidal enrolment, which is associated with a smaller number of thoracic segments, enabled faster encapsulation. This supports the hypothesis that there was a trend in the evolution of trilobites towards reduction in the number of thoracic segments in phylogenetically derived taxa in order to enhance the efficiency of enrolment.