Smooth hammerhead sharks (<Emphasis Type="Italic">Sphyrna zygaena</Emphasis>) observed off the Portuguese southern coast

Despite its worldwide distribution and vulnerable status, knowledge on the biology and ecology of the smooth hammerhead Sphyrna zygaena in the temperate NE Atlantic is very scarce. Here, we reveal intra-annual fluctuations in S. zygaena abundance in the Portuguese southwestern coast, using sightings data collected on board whale watching boats over five years (2010–14; excluding winter months). Moreover, we investigated how shark abundance is related to local environmental conditions. We describe the first smooth hammerhead “hotspot” in the NE Atlantic, and we show a recurrent pattern of occurrence during the warmer summer and autumn months (from July to October) near Sagres and Martinhal Bay and islands. Intra-annual variations in abundance were significantly associated with sea surface temperature and negatively related with upwelling index and chlorophyll a, suggesting horizontal movements linked to the seasonal changes. Hence, hammerheads moved inshore during the warmer periods (with low primary productivity), due to either direct influence of temperature in their movements or indirect influence in prey distribution. This hammerhead “hotspot” may constitute a nursery ground for S. zygaena, nevertheless, our data does not allow us to prove or refute such claim. Future telemetry-based studies should be conducted to fully understand hammerhead movements and habitat preferences, and to evaluate this “hotspot” as a critical habitat for this predator.     

Inhibition of the proton pumping ATPases of yeast and oat root plasma membranes by dicyclohexylcarbodiimide

The inhibition of the proton-pumping ATPases of yeast and oat root plasma membranes by dicyclohexylcarbodiimide (DCCD) can be correlated with the covalent incorporation of the inhibitor. Full inhibition of the yeast enzyme required the incorporation of about 1 mol DCCD/mol of the ATPase polypeptide of 100 kDa. A kinetic study of the interaction of DCCD with the yeast and oat ATPases indicates a second-order rate constant of about 500 M-1 min-1 and a stoichiometry of 1 mol DCCD/mol of enzyme, in agreement with the amount of DCCD incorporated by the yeast enzyme. It is proposed that DCCD reacts with a single carboxylic group present in a hydrophobic region of these proton-pumping ATPases and which could participate in proton binding and transport.     

Patterns in grass silicification: response to grazing history and defoliation

Summary Morphologically distinct populations of a North American perennial grass, Agropyron smithii, collected from a heavily grazed prairie dog (Cynomys ludovicianus) colony (PDC) and a grazing exclosure (EX), were grown in an environmental chamber to determine whether: (1) leaf silicon (Si) concentrations are greater in plant populations which differentiated under heavy grazing pressure, and (2) leaf silicification is inducible by defoliation. Mean shoot Si concentration of nondefoliated plants was greater in the PDC population (2.2%) than the EX population (1.9%) over the 18 wk experiment, largely as a result of differences in Si concentrations in leaf blades. However, leaf Si concentration was lower in defoliated plants of each population than in nondefoliated plants, indicating that leaf silicification was not an inducible herbivore defense mechanism in A. smithii. The higher leaf Si concentrations from the heavily grazed population may be associated with grazingrelated environmental stresses such as a warmer, drier microclimate or with morphological characteristics related to grazing tolerance or avoidance.     

Regeneration of garlic plants (allium sativum L., CV. “Chonan”) via cell culture in liquid medium

Summary Aiming at the genetic improvement of garlic cultivars, a cell suspension protocol was established which includes the induction of friable callus, establishment of cells in liquid medium, plating, regeneration, and bulb formation. Calluses of various textures from compact to friable and from green to yellowish were obtained by culturing explants excised from inner leaves of garlic bulbs on Marashig-Shoog (MS) medium with 2,4 dichlorophenoxy acetic acid (2,4-D), (1.1 mg/liter [5.0 μM]), picloram (1.2 mg/liter [5.0 μM]), and kinetin (2.1 mg/liter [10 μM]). Friable callus occurred on MS-A contained 2,4-D alone (1.0 mg/liter [4.52 μM]) and this callus was used to develop cell suspension cultures, which were maintained in liquid MS-B medium with a 2,4-D/benzyl adenine (BA) (0.5 mg/liter [2.25 μM]: 0.5 mg/liter [2.22 μM]) ratio. High plating efficiency was obtained on MS-C medium with different naphthalene acetic acid/BA combinations. Regeneration occurred after transfer of the caulogenic mass to MS-C medium containing 10 mg/liter (74.02 μM) and 20 mg/liter (148.04 μM) adenine for 60 days, followed by transfer to adenine-free medium. Plantlets transplanted to soil showed normal phenology. Shoots grown on modified MS medium supplemented with indolylbutryic acid (3.0 mg/liter [14.7 μM]) stimulated bulb formation by 30 days in culture.     

The Salmonella effector SteA binds phosphatidylinositol 4‐phosphate for subcellular targeting within host cells

Many bacterial pathogens use specialized secretion systems to deliver virulence effector proteins into eukaryotic host cells. The function of these effectors depends on their localization within infected cells, but the mechanisms determining subcellular targeting of each effector are mostly elusive. Here, we show that the Salmonella type III secretion effector SteA binds specifically to phosphatidylinositol 4‐phosphate [PI(4)P]. Ectopically expressed SteA localized at the plasma membrane (PM) of eukaryotic cells. However, SteA was displaced from the PM of Saccharomyces cerevisiae in mutants unable to synthesize the local pool of PI(4)P and from the PM of HeLa cells after localized depletion of PI(4)P. Moreover, in infected cells, bacterially translocated or ectopically expressed SteA localized at the membrane of the Salmonella‐containing vacuole (SCV) and to Salmonella‐induced tubules; using the PI(4)P‐binding domain of the Legionella type IV secretion effector SidC as probe, we found PI(4)P at the SCV membrane and associated tubules throughout Salmonella infection of HeLa cells. Both binding of SteA to PI(4)P and the subcellular localization of ectopically expressed or bacterially translocated SteA were dependent on a lysine residue near the N‐terminus of the protein. Overall, this indicates that binding of SteA to PI(4)P is necessary for its localization within host cells.     

A high-throughput fluorescence-based assay for Plasmodium dihydroorotate dehydrogenase inhibitor screening

Plasmodium dihydroorotate dehydrogenase (DHODH) is a mitochondrial membrane-associated flavoenzyme that catalyzes the rate-limiting step of de novo pyrimidine biosynthesis. DHODH is a validated target for malaria, and DSM265, a potent inhibitor, is currently in clinical trials. The enzyme catalyzes the oxidation of dihydroorotate to orotate using flavin mononucleotide (FMN) as cofactor in the first half of the reaction. Reoxidation of FMN to regenerate the active enzyme is mediated by ubiquinone (CoQ_D), which is the physiological final electron acceptor and second substrate of the reaction. We have developed a fluorescence-based high-throughput enzymatic assay to find DHODH inhibitors. In this assay, the CoQ_D has been replaced by a redox-sensitive fluorogenic dye, resazurin, which changes to a fluorescent state on reduction to resorufin. Remarkably, the assay sensitivity to find competitive inhibitors of the second substrate is higher than that reported for the standard colorimetric assay. It is amenable to 1536-well plates with Z′ values close to 0.8. The fact that the human enzyme can also be assayed in the same format opens additional applications of this assay to the discovery of inhibitors to treat cancer, transplant rejection, autoimmune diseases, and other diseases mediated by rapid cellular growth.     

Lopesia leandrae (Diptera,Cecidomyiidae), a new species of gall midge associated with Leandra ionopogon (Mart.) Cogn. (Melastomataceae), a native plant to Brazil

Lopesia leandrae (Diptera, Cecidomyiidae), a new galling species associated with Leandra ionopogon (Melastomataceae), a native plant species to Brazil, is described based on larva, pupal exuvia, male and female. L. leandrae galls were collected in an area of Atlantic Forest in Bertioga, state of São Paulo, Southeastern Brazil. The new species was compared to the other species of Lopesia.     

A new species of Youngomyia Felt from Brazil and new morphological data on Youngomyia pouteriae Maia (Insecta,Diptera, Cecidomyiidae)

Youngomyia matogrossensis Proença & Maia a new species of Cecidomyiidae (Insecta: Diptera) that induces cylindrical hairy galls on leaves of Pouteria torta (Mart.) Radlk. (Sapotaceae) is herein described and illustrated (larvae, pupal exuviae, male and female). The galler, gall and host plant were collected at Parque Nacional da Chapada dos Guimarães, in the state of Mato Grosso (Brazil). New morphological data and photographs of pupal exuviae, male and female of Youngomyia pouteriae Maia, 2001 are also provided.     

Solvent-induced free energy landscape and solute-solvent dynamic coupling in a multielement solute

Molecular dynamics simulations using a simple multielement model solute with internal degrees of freedom and accounting for solvent-induced interactions to all orders in explicit water are reported. The potential energy landscape of the solute is flat in vacuo. However, the sole untruncated solvent-induced interactions between apolar (hydrophobic) and charged elements generate a rich landscape of potential of mean force exhibiting typical features of protein landscapes. Despite the simplicity of our solute, the depth of minima in this landscape is not far in size from free energies that stabilize protein conformations. Dynamical coupling between configurational switching of the system and hydration reconfiguration is also elicited. Switching is seen to occur on a time scale two orders of magnitude longer than that of the reconfiguration time of the solute taken alone, or that of the unperturbed solvent. Qualitatively, these results are unaffected by a different choice of the water-water interaction potential. They show that already at an elementary level, solvent-induced interactions alone, when fully accounted for, can be responsible for configurational and dynamical features essential to protein folding and function.