The synthetic varespladib molecule is a multi-functional inhibitor for PLA2 and PLA2-like ophidic toxins

BackgroundThe treatment for snakebites is early administration of antivenom, which can be highly effective in inhibiting the systemic effects of snake venoms, but is less effective in the treatment of extra-circulatory and local effects. To complement standard-of-care treatments such as antibody-based antivenoms, natural and synthetic small molecules have been proposed for the inhibition of key venom components such as phospholipase A₂ (PLA₂) and PLA₂-like toxins. Varespladib (compound LY315920) is a synthetic molecule developed and clinically tested aiming to block inflammatory cascades of several diseases associated with high PLA₂s. Recent studies have demonstrated this molecule is able to potently inhibit snake venom catalytic PLA₂ and PLA₂-like toxins.MethodsIn vivo and in vitro techniques were used to evaluate the inhibitory effect of varespladib against MjTX-I. X-ray crystallography was used to reveal details of the interaction between these molecules. A new methodology that combines crystallography, mass spectroscopy and phylogenetic data was used to review its primary sequence.ResultsVarespladib was able to inhibit the myotoxic and cytotoxic effects of MjTX-I. Structural analysis revealed a particular inhibitory mechanism of MjTX-I when compared to other PLA₂-like myotoxin, presenting an oligomeric-independent function.ConclusionResults suggest the effectiveness of varespladib for the inhibition of MjTX-I, in similarity with other PLA₂ and PLA₂-like toxins.General significanceVarespladib appears to be a promissory molecule in the treatment of local effects led by PLA₂ and PLA₂-like toxins (oligomeric dependent and independent), indicating that this is a multifunctional or broadly specific inhibitor for different toxins within this superfamily.     

Assessing the landscape functional connectivity using movement maps: a case study with endemic Azorean insects

There is a vast body of literature aiming to predict, for a large number of taxa, the spatial distribution of suitable areas given the expected future changes of climatic conditions. However, such studies often overlook the role of landscape functional connectivity. This is particularly relevant for species with low vagility, as ground-dwelling insects, inhabiting areas with high human pressure due to habitat destruction and fragmentation, namely in the islands. In this study, we developed an individual-based model (IBM) that simulates individual movement according to landscape resistance and mortality probability, in order to derive the landscape movement map, and applied it to five endemic ground-dwelling insects of Terceira Island (Azores). We then confronted the movement maps of each species against the species distribution models previously developed for both current and future climatic conditions, quantifying the amount of important movement areas that are enclosed by the distribution polygons. We further sought to identify where habitat restoration would increase the overall connectivity among large habitat patches. Our results showed that, for both timeframes, the distribution models enclosed small amounts of areas predicted to be important for animal movement. Additionally, we predicted strong reductions (up to 94%) of these important areas for functional connectivity. We also identified areas in-between native forest of primary importance for restoration that may significantly increase the probability of persistence of our model species. We anticipate that this study will be useful to both conservation planners and ecologists seeking to understand species movement and dispersal both is islands and elsewhere.     

Productivity and Temperature as Drivers of Seasonal and Spatial Variations of Dissolved Methane in the Southern Bight of the North Sea

Dissolved CH₄ concentrations in the Belgian coastal zone (North Sea) ranged between 670 nmol l^?1 nearshore and 4 nmol l^?1 offshore. Spatial variations of CH₄ were related to sediment organic matter (OM) content and gassy sediments. In nearshore stations with fine sand or muddy sediments, the CH₄ seasonal cycle followed water temperature, suggesting methanogenesis control by temperature in these OM-rich sediments. In offshore stations with permeable sediments, the CH₄ seasonal cycle showed a yearly peak following the chlorophyll-a spring peak, suggesting that in these OM-poor sediments, methanogenesis depended on freshly produced OM delivery. This does not exclude the possibility that some CH₄ might originate from dimethylsulfide (DMS) or dimethylsulfoniopropionate (DMSP) or methylphosphonate transformations in the most offshore stations. Yet, the average seasonal CH₄ cycle was unrelated to those of DMS(P), very abundant during the Phaeocystis bloom. The annual average CH₄ emission was 126 mmol m^?2 y^?1 in the most nearshore stations (~4 km from the coast) and 28 mmol m^?2 y^?1 in the most offshore stations (~23 km from the coast), 1260–280 times higher than the open ocean average value (0.1 mmol m^?2 y^?1). The strong control of CH₄ by sediment OM content and by temperature suggests that marine coastal CH₄ emissions, in particular in shallow areas, should respond to future eutrophication and warming of climate. This is supported by the comparison of CH₄ concentrations at five stations obtained in March 1990 and 2016, showing a decreasing trend consistent with alleviation of eutrophication in the area.     

Terpenoids dominate the bouquet of volatile organic compounds produced by <Emphasis Type="Italic">Passiflora edulis</Emphasis> in response to herbivory by <Emphasis Type="Italic">Heliconius erato phyllis</Emphasis> (Lepidoptera: Nymphalidae)

In response to injury, plants produce volatile organic compounds (VOCs) that usually differ depending on the type of damage they have suffered (e.g., mechanical damage, herbivory, and oviposition). The objectives of this study were to identify and compare the bouquet of volatiles emitted by passion vine plants (Passiflora edulis) after injury caused by mechanical damage (MD), herbivory (HB), and oviposition (OV) by the lepidopteran, Heliconius erato phyllis. Following injury, extracts of plant emissions were collected from each treatment every 24 h for three days and were analyzed by GC and GC/MS. Results show that plants emitted 12 volatiles before and after damage, namely terpenoids, ketones, and aldehydes. Although no significant differences were detected between the three treatments individually, if the entire bouquet of volatiles is analyzed, samples collected at 24 h were different from samples collected at 48 and 72 h. However, terpenoid emission increased significantly in HB plants after 24 h. HB plants emitted approximately 6300, 50, 46, 11, 6, and 3.6 times more (3E,7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), (E)-β-ocimene, (Z)-β-farnesene, (E)-β-caryophyllene, and farnesane, respectively, compared to control plants. OV plants displayed a peak of emission of (E)-β-ocimene after 72 h, which distinguished them from HB plants. MD plants showed a general increase of VOCs versus undamaged control plants. Furthermore, it has been suggested that (E)-β-ocimene may be sequestered by larvae of H. erato phyllis as a component of the odoriferous bouquet of the abdominal scent glands present in adult males, which play a role in sexual communication.     

<Emphasis Type="Italic">Chlorella vulgaris</Emphasis> (SAG 211-12) biofilm formation capacity and proposal of a rotating flat plate photobioreactor for more sustainable biomass production

Difficulties and cost of suspended microalgal biomass harvest and processing can be overcome by cultivating microalgae as biofilms. In the present work, a new photoautotrophic biofilm photobioreactor, the rotating flat plate photobioreactor (RFPPB), was developed aiming at a cost-effective production of Chlorella vulgaris (SAG 211-12), a strain not frequently referred in the literature but promising for biofuel production. Protocols were developed for evaluating initial adhesion to different materials and testing the conditions for biofilm formation. Polyvinyl chloride substrate promoted higher adhesion and biofilm production, followed by polypropylene, polyethylene, and stainless steel. The new RFPPB was tested, aiming at optimizing incident light utilization, minimizing footprint area and simplifying biomass harvesting. Tests show that the photobioreactor is robust, promotes biofilm development, and has simple operation, small footprint, and easy biomass harvest. Biomass production (dry weight) under non-optimized conditions was 3.35 g m^?2, and areal productivity was 2.99 g m^?2 day^?1. Lipid content was 10.3% (dw), with high PUFA content. These results are promising and can be improved by optimizing some operational parameters, together with evaluation of long-term photobioreactor maximum productivity.     

<Emphasis Type="Italic">Isospora lopesi</Emphasis> n. sp. (Apicomplexa: Eimeriidae) from the eastern white-throated spadebill <Emphasis Type="Italic">Platyrinchus mystaceus</Emphasis> Vieillot (Passeriformes: Tyranni: Tyrannidae) in South America

A species of Isospora Schneider, 1881 (Protozoa: Apicomplexa: Eimeriidae) considered as new to science is described and characterised molecularly from the eastern white-throated spadebill Platyrinchus mystaceus Vieillot in the Parque Nacional do Itatiaia, southeastern Brazil. Isospora lopesi n. sp. has oöcysts that are subspheroidal to ovoidal, 18–24 × 18–22 (20.6 × 19.7) µm, with smooth, bilayered wall, c.1.5 μm thick. Micropyle and oöcyst residuum are absent, but one polar granule is present. Sporocysts are ellipsoidal, 12–16 × 8–11 (14.4 × 8.6) µm. The Stieda body is flattened to half-moon-shaped and sub-Stieda body is rounded. Sporocyst residuum is present, consisting of numerous spherules of different sizes. Sporozoites are vermiform with anterior and posterior refractile bodies and nucleus. Molecular analysis was conducted at the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. This new isolate exhibited similarity greater than 98% with Isospora spp. isolates from spectacled warblers Sylvia conspicillata Temminck, 1820. This is the fourth isosporoid coccidian described from New World tyrannid birds, but is the first to have a complementary molecular characterisation.     

Role of Nγ-Acetyldiaminobutyrate as an Enzyme Stabilizer and an Intermediate in the Biosynthesis of Hydroxyectoine

Strain CHR63 is a salt-sensitive mutant of the moderately halophilic wild-type strain Halomonas elongata DSM 3043 that is affected in the ectoine synthase gene (ectC). This strain accumulates large amounts of Nγ-acetyldiaminobutyrate (NADA), the precursor of ectoine (D. Cánovas, C. Vargas, F. Iglesias-Guerra, L. N. Csonka, D. Rhodes, A. Ventosa, and J. J. Nieto, J. Biol. Chem. 272:25794–25801, 1997). Hydroxyectoine, ectoine, and glucosylglycerate were also identified by nuclear magnetic resonance (NMR) as cytoplasmic organic solutes in this mutant. Accumulation of NADA, hydroxyectoine, and ectoine was osmoregulated, whereas the levels of glucosylglycerate decreased at higher salinities. The effect of the growth stage on the accumulation of solutes was also investigated. NADA was purified from strain CHR63 and was shown to protect the thermolabile enzyme rabbit muscle lactate dehydrogenase against thermal inactivation. The stabilizing effect of NADA was greater than the stabilizing effect of ectoine or potassium diaminobutyrate. A ¹H NMR analysis of the solutes accumulated by the wild-type strain and mutants CHR62 (ectA::Tn1732) and CHR63 (ectC::Tn1732) indicated that H. elongata can synthesize hydroxyectoine by two different pathways—directly from ectoine or via an alternative pathway that converts NADA into hydroxyectoine without the involvement of ectoine.