Differentiation of Human Umbilical Cord Matrix Mesenchymal Stem Cells into Neural-Like Progenitor Cells and Maturation into an Oligodendroglial-Like Lineage

Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton''s jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.     

Size‐correlated morpho‐physiology of the aroid vine Rhodospatha oblongata along a vertical gradient in a Brazilian rain forest

In this work, we analyse morpho‐physiological modifications presented during the allomorphic growth of the aroid vine Rhodospatha oblongata Poepp throughout its ascent into the forest canopy. We test the hypothesis that morphological modifications in the root, shoot and leaf are followed by a gradual improvement of the xylem vascular system in order to increase water acquisition and transport as body size increases. The characterisation of these structural modifications was based on 30–35 specimens divided into six size classes. The dimensions of shoots, leaves and roots were quantified and qualified. The transition from the terrestrial to the epiphytic phase was followed by a simultaneous increase of leaf number and lamina area, together with increased length and diameter of the petiole. Furthermore, as the plant grows, the shoot internodes become shorter and thicker. However, occurrence of aerial roots is the most important characteristic in the ascending phase. In taller individuals, the increase in number of roots with wider xylem vessels guarantees an increased theoretical xylem hydraulic conductance for this growth phase. Along an acropetal direction of the same shoot, the diameter of xylem vessels increased, while the number of vessels per stele area decreased, in contrast with such allometric models as that of West, Brown and Enquist, showing that xylem vessel number and diameter taper in a reverse manner along the same direction. Such structural changes of R. oblongata allow improved foraging for light and water, facilitating the survival of bigger‐sized plants of this vine in the canopy.     

Adaptive foraging of leaf‐cutter ants to spatiotemporal changes in resource availability in Neotropical savannas

1. Generalist herbivores feed on a wide and diverse set of species, but fine‐scale foraging patterns may be affected by the interplay between the quality, quantity and spatial distribution of host plants. 2. The foraging patterns of a prevalent Neotropical herbivore, the leaf‐cutter ant Atta laevigata, in the Brazilian Cerrado savannas were examined in order to determine if patterns observed are in concert with central‐place foraging predictions. 3. The results showed that A. laevigata acts as a polyphagous but highly selective herbivore, with ant attacks often resulting in partial defoliation of less‐preferred species and full defoliation of preferred ones. It was found, for the first time, that there is a strong and positive relationship between the relative attack frequency on plants from preferred species and foraging distance to the nest. This suggests a balance between the quality of plant resources harvested and costs involved in their transportation. It was also observed that colonies focused their harvest on preferred species in months with low availability of young leaves. Consequently, high herbivory rate was more frequent in plants attacked far away from the nest and in dry months. 4. These assessments highlight the fact that Atta colonies may become more selective as foraging distance to the nest increases and in response to fluctuations in the availability of palatable resources throughout the year. The results also show some dissimilarities in the foraging behaviours of A. laevigata when compared with other locations, suggesting that widely distributed herbivores may modify foraging strategies across their geographic range.     

Functional specialization and differential regulation of short‐chain carboxylic acid transporters in the pathogen Candida albicans

The major fungal pathogen Candida albicans has the metabolic flexibility to assimilate a wide range of nutrients in its human host. Previous studies have suggested that C. albicans can encounter glucose‐poor microenvironments during infection and that the ability to use alternative non‐fermentable carbon sources contributes to its virulence. JEN1 encodes a monocarboxylate transporter in C. albicans and we show that its paralogue, JEN2, encodes a novel dicarboxylate plasma membrane transporter, subjected to glucose repression. A strain deleted in both genes lost the ability to transport lactic, malic and succinic acids by a mediated mechanism and it displayed a growth defect on these substrates. Although no significant morphogenetic or virulence defects were found in the double mutant strain, both JEN1 and JEN2 were strongly induced during infection. Jen1‐GFP (green fluorescent protein) and Jen2‐GFP were upregulated following the phagocytosis of C. albicans cells by neutrophils and macrophages, displaying similar behaviour to an Icl1‐GFP fusion. In the murine model of systemic candidiasis approximately 20–25% of C. albicans cells infecting the kidney expressed Jen1‐GFP and Jen2‐GFP. Our data suggest that Jen1 and Jen2 are expressed in glucose‐poor niches within the host, and that these short‐chain carboxylic acid transporters may be important in the early stages of infection.     

Simultaneous bidirectional magnesium ion flux measurements in single barnacle muscle cells by mass spectrometry.

Stable isotopes of Mg were used to measure bidirectional magnesium ion fluxes in single barnacle giant muscle fibers immersed in Ca- and Na-free, isosmotic media. Measurements were made using a mass spectrometric technique, thermal ionization mass spectrometry (TIMS), in conjunction with atomic absorption spectroscopy. Kinetic relations based on a first-order model were developed that permit the determination of unidirectional rate coefficients for Mg influx, ki, and efflux, ke, in the same experiment from knowledge of initial conditions and the initial and final ratios of 26Mg/24Mg and 25Mg/24Mg in ambient solutions (i.e., by isotope dilution). Such determinations were made for three values of the external Mg ion concentration: 5, 25, and 60 mM. At the concentration [Mg+2]o = 5 mM, ki and ke were about equal at a value of 0.01 min-1. At the higher values of [Mg+2]o, the values of ke increased along a curve suggesting saturation, whereas the values of ki remained essentially constant. As could be expected on the basis of a constant ki, the initial influx rate varied in direct linear proportion to [Mg+2]o, and was 11.8 pmol/cm2s when [Mg+2]o was 5 mM. However, the initial efflux rate appeared to increase nonlinearly with [Mg+2]o, varying from 13.4 pmol/cm2s ([ Mg+2]o = 5 mM) to approximately 80 pmol/cm2s ([ Mg+2]o = 60 mM). The results are consistent with a model that assumes Mg influx to be mainly an electrodiffusive inward leak with PMg = 0.07 cm/s and Mg efflux to be almost entirely by active transport processes. Where comparisons can be made, the rate coefficients determined from stable isotope measurements agree with those previously obtained using radioactive Mg. The rate coefficients can be used to correctly predict time-dependent changes in total fiber Mg content. The results support the conclusion that nonradioactive tracers can be used to measure ion fluxes and ion flux ratios in excitable cells; it is expected that this method will greatly assist in the study of Mg regulation in general.     

Identification of Novel Loci Associated with Gastrointestinal Parasite Resistance in a Red Maasai x Dorper Backcross Population

Gastrointestinal (GI) parasitic infection is the main health constraint for small ruminant production, causing loss of weight and/or death. Red Maasai sheep have adapted to a tropical environment where extreme parasite exposure is a constant, especially with highly pathogenic Haemonchus contortus. This breed has been reported to be resistant to gastrointestinal parasite infection, hence it is considered an invaluable resource to study associations between host genetics and resistance. The aim of this study was to identify polymorphisms strongly associated with host resistance in a double backcross population derived from Red Maasai and Dorper sheep using a SNP-based GWAS analysis. The animals that were genotyped represented the most resistant and susceptible individuals based on the tails of phenotypic distribution (10% each) for average faecal egg counts (AVFEC). AVFEC, packed cell volume (AVPCV), and live weight (AVLWT) were adjusted for fixed effects and co-variables, and an association analysis was run using EMMAX. Revised significance levels were calculated using 100,000 permutation tests. The top five significant SNP markers with - log10 p-values >3.794 were observed on five different chromosomes for AVFEC, and BLUPPf90/PostGSf90 results confirmed EMMAX significant regions for this trait. One of these regions included a cluster of significant SNP on chromosome (Chr) 6 not in linkage disequilibrium to each other. This genomic location contains annotated genes involved in cytokine signalling, haemostasis and mucus biosynthesis. Only one association detected on Chr 7 was significant for both AVPCV and AVLWT. The results generated here reveal candidate immune variants for genes involved in differential response to infection and provide additional SNP marker information that has potential to aid selection of resistance to gastrointestinal parasites in sheep of a similar genetic background to the double backcross population.     

DNA Barcoding of Neotropical Sand Flies (Diptera,Psychodidae, Phlebotominae): Species Identification and Discovery within Brazil

DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23–19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.