Assessment of different sperm functional tests in golden‐headed lion tamarins (Leontopithecus chrysomelas)

The golden‐headed lion tamarin (Leontopithecus chrysomelas) is an endangered species endemic to Brazil's Atlantic Forest, a shrinking biodiversity hotspot. As in other Neotropical primates, its semen characteristics and freezability are poorly studied. Hence, reproductive technologies for callitrichids would greatly benefit from reliable methods of semen analysis. In a bid to promote reproductive research in tamarins, we validated simple and inexpensive sperm function tests that can be used to monitor sperm‐egg binding, plasma membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. Ejaculates from adult males were individually diluted and divided into control and damage‐induced aliquots, and then samples comprising assorted amounts of damaged spermatozoa were examined by organelle‐specific tests. Our findings showed that sperm‐binding in chicken egg perivitelline membrane (EPM) positively correlated with the number of spermatozoa injured by snap‐freezing. Eosin‐nigrosin (EN) and propidium iodide readings were correlated with each other, and both provided robust measurements of plasma membrane integrity. A high correlation between expected and measured amounts of acrosome‐intact spermatozoa was found using Fast Green‐Rose Bengal (FG‐RB), Coomassie Blue (CB), and FITC‐PSA stains, and all three methods exhibited comparable results. Likewise, different percentages of UV‐irradiated spermatozoa were accurately assessed for DNA integrity by Toluidine Blue (TB) and sperm chromatin dispersion (SCD) tests. Comparisons between 3,3′‐diaminobenzidine (DAB) and JC‐1 stains also indicated the reliability of the former assay to ascertain gradual increases in spermatozoa with greater mitochondrial function. These data confirmed that different parts of the tamarin spermatozoa can be simply and consistently evaluated by EPM, EN, FG‐RB, CB, TB, and DAB protocols.     

Design and synthesis of 4,5,6,7‐tetrahydro‐1H‐1,2‐diazepin‐7‐one derivatives as a new series of Phosphodiesterase 4 (PDE4) inhibitors

Phosphodiesterase 4 (PDE4) inhibitors have attractive therapeutic potential in respiratory, inflammatory, metabolic and CNS disorders. The present work details the design, chemical exploration and biological profile of a novel PDE4 inhibitor chemotype. A diazepinone ring was identified as an under-represented heterocyclic system fulfilling a set of PDE4 structure-based design hypotheses. Rapid exploration of the structure activity relationships for the series was enabled by robust and scalable two/three-steps parallel chemistry protocols. The resulting compounds demonstrated PDE4 inhibitory activity in cell free and cell-based assays comparable to the Zardaverine control used, suggesting potential avenues for their further development.     

Tripartite efflux pumps: energy is required for dissociation,but not assembly or opening of the outer membrane channel of the pump

The MtrCDE multidrug pump, from Neisseria gonorrhoeae, is assembled from the inner and outer membrane proteins MtrD and MtrE, which are connected by the periplasmic membrane fusion protein MtrC. Although it is clear that MtrD delivers drugs to the channel of MtrE, it remains unclear how drug delivery and channel opening are connected. We used a vancomycin sensitivity assay to test for opening of the MtrE channel. Cells expressing MtrE or MtrE‐E434K were insensitive to vancomycin; but became moderately and highly sensitive to vancomycin respectively, when coexpressed with MtrC, suggesting that the MtrE channel opening requires MtrC binding and is energy‐independent. Cells expressing wild‐type MtrD, in an MtrCE background, were vancomycin‐insensitive, but moderately sensitive in an MtrCE‐E434K background. The mutation of residues involved in proton translocation inactivated MtrD and abolished drug efflux, rendered both MtrE and MtrE‐E434K vancomycin‐insensitive; imply that the pump–component interactions are preserved, and that the complex is stable in the absence of proton flux, thus sealing the open end of MtrE. Following the energy‐dependent dissociation of the tripartite complex, the MtrE channel is able to reseal, while MtrE‐E434K is unable to do so, resulting in the vancomycin‐sensitive phenotype. Thus, our findings suggest that opening of the OMP via interaction with the MFP is energy‐independent, while both drug export and complex dissociation require active proton flux.     

<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.     

Inundation,Hydrodynamics and Vegetation Influence Carbon Dioxide Concentrations in Amazon Floodplain Lakes

Ecosystems - Extensive floodplains and numerous lakes in the Amazon basin are well suited to examine the role of floodable lands within the context of the sources and processing of carbon within...     

Production of Lactic Acid Enantiomers by Lactobacillus Strains under Limited Dissolved Oxygen Conditions in the Presence of a Pentose Fraction

Applied Biochemistry and Microbiology - Lactic acid has a privileged position in the family of carboxylic substances due to its several applications, including the production of biodegradable...     

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.