Biology of Tupiocoris cucurbitaceus (Hemiptera: Miridae), a predator of the greenhouse whitefly Trialeurodes vaporariorum (Hemiptera: Aleyrodidae) in tomato crops in Argentina

The predator Tupiocoris cucurbitaceus is frequently found attacking Trialeurodes vaporariorum in greenhouses without pesticide applications in Argentina. The objective of these studies was to evaluate some biological characteristics of this species fed on three types of diet (whitefly nymphs, Sitotroga cerealella eggs and a mix of both) and on two host plants (tomato and tobacco), under controlled experimental conditions. Preimaginal developmental time for female and male bugs was shorter in the presence of whiteflies than with only moth eggs. Females lived longer when they ate only whitefly nymphs compared to a mixed diet or only moth eggs. The amount of adult descendants was greater when bugs could eat whiteflies, regardless of the presence of S. cerealella. Embryonic development time, male longevity and sex proportion were not affected by the diet or the host plant. Prey consumption was evaluated for three T. cucurbitaceus life history stages (fourth/fifth instar nymphs, female and male adults) on two types of prey (whitefly nymphs and S. cerealella eggs). On tomato, females were more voracious than males and nymphs. On tobacco, adults and nymphs consumed more S. cerealella than T.vaporariorum nymphs, but again, bug females preyed more than males and nymphs. Results demonstrate that T. cucurbitaceus can survive, develop and reproduce normally using both T. vaporariorum and S. cerealella eggs as prey on tobacco or tomato plants. This information can be useful for managing this predator against T. vaporariorum through conservative or augmentative biological control strategies.     

Communicating old-growth forest through an educational trail

This paper describes the experience of developing an educational trail inside an old-growth beech forest in central Italy. In the last 150 years local people exploited this stand almost exclusively for recreational purposes and now the beechwood has acquired new ecological value after having reached old-growth status. Despite the cultural-historical importance of the site, there is a substantial lack of knowledge about this magnificent stand among members of the public, who have no tools to understand its dynamics nor to accept those structural changes related to its natural evolution. Tourist facilities were restored at the site, according to non-restricting criteria, and an educational program was developed in order to promote a shared comprehension of old-growth forests, their ecological processes and value, the services they provide, and to increase tourist awareness toward this fragile ecosystem. We discuss the educational role ecotourism might potentially hold in western developed countries to link the public with the scientific world, educating people about nature, while preventing negative impacts on natural environments. Even if the importance of local communities in conservation of natural resources has been widely recognized by a number of conservationists, public involvement, particularly in forest conservation and management, still represents a difficult challenge, since it requires the implementation of a common understanding about forest ecosystems, forest goods and forest services. The educational trail inside the beech forest of Monte Cimino may represent a low impact strategy for preserving forest aesthetic functionality, limiting potential damages to the forest while increasing its importance for nature conservation.     

Revisiting the Acanthamoeba species that form star-shaped cysts (genotypes T7, T8, T9, and T17): characterization of seven new Brazilian environmental isolates and phylogenetic inferences

Free-living amoebae of the genus Acanthamoeba are the agents of both opportunistic and non-opportunistic infections and are frequently isolated from the environment. Of the 17 genotypes (T1-T17) identified thus far, 4 (T7, T8, T9, and T17) accommodate the rarely investigated species of morphological group I, those that form large, star-shaped cysts. We report the isolation and characterization of 7 new Brazilian environmental Acanthamoeba isolates, all assigned to group I. Phylogenetic analyses based on partial (~1200 bp) SSU rRNA gene sequences placed the new isolates in the robustly supported clade composed of the species of morphological group I. One of the Brazilian isolates is closely related to A. comandoni (genotype T9), while the other 6, together with 2 isolates recently assigned to genotype T17, form a homogeneous, well-supported group (2·0% sequence divergence) that likely represents a new Acanthamoeba species. Thermotolerance, osmotolerance, and cytophatic effects, features often associated with pathogenic potential, were also examined. The results indicated that all 7 Brazilian isolates grow at temperatures up to 40°C, and resist under hyperosmotic conditions. Additionally, media conditioned by each of the new Acanthamoeba isolates induced the disruption of SIRC and HeLa cell monolayers.     

Water availability modifies tolerance to photo-oxidative pollutants in transplants of the lichen Flavoparmelia caperata

The hypothesis that a daily water supply allows a lichen to endure the negative effects of environmental concentrations of NO_x and O₃ was tested with a transplant experiment. Five groups (0, A–D) of Flavoparmelia caperata samples derived from the same thalli were used for destructive, pre-exposure measurements (0), or exposed for 5 weeks in the rural collection site (A), and in a urban site with high levels of NO_x and O₃ (B–D). Two groups (C, D) were daily watered half an hour before the daily peak of NO_x (C), and O₃ (D). The comparison between pre- and post-exposure measurements of stress biomarkers revealed that the different thallus hydration regime modified the pollution tolerance as well as the physiology of the exposed samples. The non-watered group B suffered an evident decrease in F_v/F_m and reduced glutathione, but increased ion leakage, whereas the watered groups C and D showed only decreased non-photosynthetic-quenching, possibly derived from NO_x exposure. Ozone, which was higher in the rural than in the urban site, did not significantly affect the lichen metabolism. Our results re-open the discussion on the so-called “drought hypothesis”, which suggests that the lichen desert observed in urban areas of central and eastern Europe is more a matter of dry microclimate than of air pollution.     

Transient interactions of a slow-folding protein with the Hsp70 chaperone machinery

Most known proteins have at least one local Hsp70 chaperone binding site. Does this mean that all proteins interact with Hsp70 as they fold? This study makes an initial step to address the above question by examining the interaction of the E.coli Hsp70 chaperone (known as DnaK) and its co-chaperones DnaJ and GrpE with a slow-folding E.coli substrate, RNase H^D. Importantly, this protein is a nonobligatory client, and it is able to fold in vitro even in the absence of chaperones. We employ stopped-flow mixing, chromatography, and activity assays to analyze the kinetic perturbations induced by DnaK/DnaJ/GrpE (K/J/E) on the folding of RNase H^D. We find that K/J/E slows down RNase H^D''s apparent folding, consistent with the presence of transient chaperone-substrate interactions. However, kinetic retardation is moderate for this slow-folding client and it is expected to be even smaller for faster-folding substrates. Given that the interaction of folding-competent substrates such as RNase H^D with the K/J/E chaperones is relatively short-lived, it does not significantly interfere with the timely production of folded biologically active substrate. The above mode of action is important because it preserves K/J/E bioavailability, enabling this chaperone system to act primarily by assisting the folding of other misfolded and (or) aggregation-prone cellular proteins that are unable to fold independently. When refolding is carried out in the presence of K/J and absence of the nucleotide exchange factor GrpE, some of the substrate population becomes trapped as a chaperone-bound partially unfolded state.     

Long-term decrease in VLA-4 expression and functional impairment of dendritic cells during natalizumab therapy in patients with multiple sclerosis

Myeloid and plasmacytoid dendritic cells (mDCs, pDCs) are central to the initiation and the regulation of immune processes in multiple sclerosis (MS). Natalizumab (NTZ) is a humanized monoclonal antibody approved for the treatment of MS that acts by blocking expression of VLA-4 integrins on the surface of leukocytes. We determined the proportions of circulating DC subsets and analyzed expression of VLA-4 expression in 6 relapsing-remitting MS patients treated with NTZ for 1 year. VLA-4 expression levels on pDCs and mDCs decreased significantly during follow-up. In vitro coculture of peripheral blood mononuclear cells and pDCs, with different doses of NTZ in healthy controls (HC) and MS patients showed dose-dependent down-regulation of VLA-4 expression levels in both MS patients and HC, and reduced functional ability to stimulate antigen-specific T-lymphocyte responses. The biological impact of NTZ may in part be attributable to inhibition of transmigration of circulating DCs into the central nervous system, but also to functional impairment of interactions between T cells and DC.     

CNF1 improves astrocytic ability to support neuronal growth and differentiation in vitro

Modulation of cerebral Rho GTPases activity in mice brain by intracerebral administration of Cytotoxic Necrotizing Factor 1 (CNF1) leads to enhanced neurotransmission and synaptic plasticity and improves learning and memory. To gain more insight into the interactions between CNF1 and neuronal cells, we used primary neuronal and astrocytic cultures from rat embryonic brain to study CNF1 effects on neuronal differentiation, focusing on dendritic tree growth and synapse formation, which are strictly modulated by Rho GTPases. CNF1 profoundly remodeled the cytoskeleton of hippocampal and cortical neurons, which showed philopodia-like, actin-positive projections, thickened and poorly branched dendrites, and a decrease in synapse number. CNF1 removal, however, restored dendritic tree development and synapse formation, suggesting that the toxin can reversibly block neuronal differentiation. On differentiated neurons, CNF1 had a similar effacing effect on synapses. Therefore, a direct interaction with CNF1 is apparently deleterious for neurons. Since astrocytes play a pivotal role in neuronal differentiation and synaptic regulation, we wondered if the beneficial in vivo effect could be mediated by astrocytes. Primary astrocytes from embryonic cortex were treated with CNF1 for 48 hours and used as a substrate for growing hippocampal neurons. Such neurons showed an increased development of neurites, in respect to age-matched controls, with a wider dendritic tree and a richer content in synapses. In CNF1-exposed astrocytes, the production of interleukin 1β, known to reduce dendrite development and complexity in neuronal cultures, was decreased. These results demonstrate that astrocytes, under the influence of CNF1, increase their supporting activity on neuronal growth and differentiation, possibly related to the diminished levels of interleukin 1β. These observations suggest that the enhanced synaptic plasticity and improved learning and memory described in CNF1-injected mice are probably mediated by astrocytes.     

CD14 deficiency impacts glucose homeostasis in mice through altered adrenal tone

The toll-like receptors comprise one of the most conserved components of the innate immune system, signaling the presence of molecules of microbial origin. It has been proposed that signaling through TLR4, which requires CD14 to recognize bacterial lipopolysaccharide (LPS), may generate low-grade inflammation and thereby affect insulin sensitivity and glucose metabolism. To examine the long-term influence of partial innate immune signaling disruption on glucose homeostasis, we analyzed knockout mice deficient in CD14 backcrossed into the diabetes-prone C57BL6 background at 6 or 12 months of age. CD14-ko mice, fed either normal or high-fat diets, displayed significant glucose intolerance compared to wild type controls. They also displayed elevated norepinephrine urinary excretion and increased adrenal medullary volume, as well as an enhanced norepinephrine secretory response to insulin-induced hypoglycemia. These results point out a previously unappreciated crosstalk between innate immune- and sympathoadrenal- systems, which exerts a major long-term effect on glucose homeostasis.