Length–weight relationships for estuarine fishes in South Brazil

The present study reports the length‐weight relationships (LWRs) for 18 estuarine species sampled in Guaratuba Bay, South Brazil. For the majority of these 18 species the LWRs are known from the literature, but these estimates do not consider sexes separately which is done in this contribution. Different fishing gears were used in the samplings (cylindrical nets – 1 mm mesh size, bottom trawls – 250 mm mesh size in the arms and 12 mm mesh size in the bag, beach seine – 5 mm mesh size and hook and line fishery) between September 2007 to April 2014. The specimens were sexed and the LWRs were calculated for each sex separately. There were differences between sexes in growth type, showing the importance of sexing in these studies. We reported for the first time (based on FishBase) the LWRs for two species: Ctenogobius shufeldti and Hypleurochilus fissicornis.     

A new species of Cambeva (Siluriformes,Trichomycteridae) from the Rio Ivaí basin,Upper Rio Paraná basin,Paraná State,Brazil

A new species of Cambeva endemic to the Rio Ivaí basin, Upper Paraná basin, is described combining morphological and molecular data. This new species is distinguished from all congeners by characters related to the number of pectoral-fin rays, to the colour pattern of the dorsal and lateral surface of the body, to the presence of diffuse blotches in the ventral surface of body, to the presence of a pelvic-fin and pelvic girdle, to the number of odontodes in the inter-opercular and opercular patches, to the number of dorsal and ventral procurrent rays. In addition, the analysis of mitochondrial DNA sequences identified a satisfactory genetic distance between this new species and its congeners. The new species from the Rio Ivaí reinforces its characteristics as an area of endemism for fishes in the Upper Rio Paraná basin.     

The Effects of Spatial Scale on Breakdown of Leaves in a Tropical Watershed

The objective was to assess the effects of natural variation in the physical structure of the environment on biological communities and on the processing of Eucalyptus cloeziana and Inga laurina and to identify the controlling factors at different scales along stream order gradients. The study area consisted of 14 sampling sites distributed within a tropical watershed (1st, 2nd, 3rd and 4th order streams replicated in 4 sub-basins). Our samples consisted of 3 g of leaves of E. cloeziana (high-quality) and I. laurina (low-quality) placed in 252 bags with 10mm mesh (measured by the chemical composition of the detritus). Four samples of each leaf type were collected periodically (three times) over a period of 75–125 days and washed on a sieve to separate the invertebrates. A series of leaf disks were cut to determine ash-free dry mass, polyphenol, lignin, cellulose, total microbial biomass and fungal biomass, and the remaining material was oven-dried to determine the dry weight. We performed analyses within and between spatial scales (regional and local) to assess which watershed scale was the more import determinant of the leaf breakdown rate (k). The microbial and shredder were most influenced at the local scale (stream order). Shredders were influenced by microorganisms, with stronger interactions between them than were found to drive the k at the local scale. Moreover, differences in the overall k and abiotic variables were more strongly influenced at the regional scale (sub-basin), showing that the study scale alters the response of the studied variables. We found higher k values at higher values of water velocity, dissolved oxygen and temperature, all of which accelerate biological metabolism in response to variations on the regional scale. Watersheds with warmer microclimates and streams with higher nutrient levels and oxygen could be accelerating the ecosystem metabolism, independent of the detritus quality.     

Probing the interaction between heparan sulfate proteoglycan with biologically relevant molecules in mimetic models for cell membranes: A Langmuir film study

Investigating the role of proteoglycans associated to cell membranes is fundamental to comprehend biochemical process that occurs at the level of membrane surfaces. In this paper, we exploit syndecan-4, a heparan sulfate proteoglycan obtained from cell cultures, in lipid Langmuir monolayers at the air-water interface. The monolayer served as a model for half a membrane, and the molecular interactions involved could be evaluated with tensiometry and vibrational spectroscopy techniques. Polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS) employed in a constant surface pressure regime showed that the main chemical groups for syndecan-4 were present at the air-water interface. Subsequent monolayer decompression and compression showed surface pressure-area isotherms with a large expansion for the lipid monolayers interacting with the cell culture reported to over-express syndecan-4, which was also an indication that the proteoglycan was inserted in the lipid monolayer. The introduction of biological molecules with affinity for syndecam-4, such as growth factors, which present a key role in biochemical process of cell signaling, changed the surface properties of the hybrid film, leading to a model, by which the growth factor binds to the sulfate groups present in the heparan sulfate chains. The polypeptide moiety of syndecan-4 responds to this interaction changing its conformation, which leads to lipid film relaxation and further monolayer condensation.     

The natural variation of a neural code

The way information is represented by sequences of action potentials of spiking neurons is determined by the input each neuron receives, but also by its biophysics, and the specifics of the circuit in which it is embedded. Even the "code" of identified neurons can vary considerably from individual to individual. Here we compared the neural codes of the identified H1 neuron in the visual systems of two families of flies, blow flies and flesh flies, and explored the effect of the sensory environment that the flies were exposed to during development on the H1 code. We found that the two families differed considerably in the temporal structure of the code, its content and energetic efficiency, as well as the temporal delay of neural response. The differences in the environmental conditions during the flies' development had no significant effect. Our results may thus reflect an instance of a family-specific design of the neural code. They may also suggest that individual variability in information processing by this specific neuron, in terms of both form and content, is regulated genetically.     

Downregulation of the cAMP/PKA Pathway in PC12 Cells Overexpressing NCS-1

It is well known that dopamine imbalances are associated with many psychiatric disorders and that the dopaminergic receptor D₂ is the main target of antipsychotics. Recently it was shown that levels of two proteins implicated in dopaminergic signaling, Neuronal calcium sensor-1 (NCS-1) and DARPP-32, are altered in the prefrontal cortex (PFC) of both schizophrenic and bipolar disorder patients. NCS-1, which inhibits D₂ internalization, is upregulated in the PFC of both patients. DARPP-32, which is a downstream effector of dopamine signaling, integrates the pathways of several neurotransmitters and is downregulated in the PFC of both patients. Here, we used PC12 cells stably overexpressing NCS-1 (PC12-NCS-1 cells) to address the function of this protein in DARPP-32 signaling pathway in vitro. PC12-NCS-1 cells displayed downregulation of the cAMP/PKA pathway, with decreased levels of cAMP and phosphorylation of CREB at Ser133. We also observed decreased levels of total and phosphorylated DARPP-32 at Thr34. However, these cells did not show alterations in the levels of D₂ and phosphorylation of DARPP-32 at Thr75. These results indicate that NCS-1 modulates PKA/cAMP signaling pathway. Identification of the cellular mechanisms linking NCS-1 and DARPP-32 may help in the understanding the signaling machinery with potential to be turned into targets for the treatment of schizophrenia and other debilitating psychiatric disorders.     

Muscle-type 6-phosphofructo-1-kinase and aldolase associate conferring catalytic advantages for both enzymes

6-Phosphofructo-1-kinase (PFK) and aldolase are two sequential glycolytic enzymes that associate forming heterotetramers containing a dimer of each enzyme. Although free PFK dimers present a negligible activity, once associated to aldolase these dimers are as active as the fully active tetrameric conformation of the enzyme. Here we show that aldolase-associated PFK dimers are not inhibited by clotrimazole, an antifungal azole derivative proposed as an antineoplastic drug due to its inhibitory effects on PFK. In the presence of aldolase, PFK is not modulated by its allosteric activators, ADP and fructose-2,6-bisphosphate, but is still inhibited by citrate and lactate. The association between the two enzymes also results on the twofold stimulation of aldolase maximal velocity and affinity for its substrate. These results suggest that the association between PFK and aldolase confers catalytic advantage for both enzymes and may contribute to the channeling of the glycolytic metabolism.     

Adhesins of Leptospira interrogans Mediate the Interaction to Fibrinogen and Inhibit Fibrin Clot Formation In Vitro

We report in this work that Leptospira strains, virulent L. interrogans serovar Copenhageni, attenuated L. interrogans serovar Copenhageni and saprophytic L. biflexa serovar Patoc are capable of binding fibrinogen (Fg). The interaction of leptospires with Fg inhibits thrombin- induced fibrin clot formation that may affect the haemostatic equilibrium. Additionally, we show that plasminogen (PLG)/plasmin (PLA) generation on the surface of Leptospira causes degradation of human Fg. The data suggest that PLA-coated leptospires were capable to employ their proteolytic activity to decrease one substrate of the coagulation cascade. We also present six leptospiral adhesins and PLG- interacting proteins, rLIC12238, Lsa33, Lsa30, OmpL1, rLIC11360 and rLIC11975, as novel Fg-binding proteins. The recombinant proteins interact with Fg in a dose-dependent and saturable fashion when increasing protein concentration was set to react to a fix human Fg concentration. The calculated dissociation equilibrium constants (K_D) of these reactions ranged from 733.3±276.8 to 128±89.9 nM for rLIC12238 and Lsa33, respectively. The interaction of recombinant proteins with human Fg resulted in inhibition of fibrin clot by thrombin-catalyzed reaction, suggesting that these versatile proteins could mediate Fg interaction in Leptospira. Our data reveal for the first time the inhibition of fibrin clot by Leptospira spp. and presents adhesins that could mediate these interactions. Decreasing fibrin clot would cause an imbalance of the coagulation cascade that may facilitate bleeding and help bacteria dissemination     

Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae

Objectives

Develop a Cell Surface Display system in Saccharomyces cerevisiae, based on the construction of an expression cassette for pYES2 plasmid.

Results

The construction of an expression cassette containing the α-factor signal peptide and the C-terminal portion of the α-agglutinin protein was made and its sequence inserted into a plasmid named pYES2/gDαAgglutinin. The construction allows surface display of bovine herpesvirus type 5 (BoHV-5) glycoprotein D (gD) on S. cerevisiae BY4741 strain. Recombinant protein expression was confirmed by dot blot, and indirect immunofluorescence using monoclonal anti-histidine antibodies and polyclonal antibodies from mice experimentally vaccinated with a recombinant gD.

Conclusions

These results demonstrate that the approach and plasmid used represent not only an effective system for immobilizing proteins on the yeast cell surface, as well as a platform for immunobiologicals development.