Cloning, Expression, Purification and Characterization of His-Tagged Human Glucose-6-Phosphate Dehydrogenase: A Simplified Method for Protein Yield

Glucose-6-phosphate dehydrogenase (G6PD) catalyzes the first step of the pentose phosphate pathway. In erythrocytes, the functionality of the pathway is crucial to protect these cells against oxidative damage. G6PD deficiency is the most frequent enzymopathy in humans with a global prevalence of 4.9 %. The clinical picture is characterized by chronic or acute hemolysis in response to oxidative stress, which is related to the low cellular activity of G6PD in red blood cells. The disease is heterogeneous at genetic level with around 160 mutations described, mostly point mutations causing single amino acid substitutions. The biochemical studies aimed to describe the detrimental effects of mutations on the functional and structural properties of human G6PD are indispensable to understand the molecular physiopathology of this disease. Therefore, reliable systems for efficient expression and purification of the protein are highly desirable. In this work, human G6PD was heterologously expressed in Escherichia coli and purified by immobilized metal affinity chromatography in a single chromatographic step. The structural and functional characterization indicates that His-tagged G6PD resembles previous preparations of recombinant G6PD. In contrast with previous protein yield systems, our method is based on commonly available resources and fully accessible laboratory equipment; therefore, it can be readily implemented.     

Gadolinium-containing carbon nanomaterials for magnetic resonance imaging: Trends and challenges

Gadolinium-containing carbon nanomaterials are a new class of contrast agent for magnetic resonance imaging. They are characterized by a superior proton relaxivity to any current commercial gadolinium contrast agent and offer the possibility to design multifunctional contrasts. Intense efforts have been made to develop these nanomaterials because of their potential for better results than the available gadolinium contrast agents. The aim of the present work is to provide a review of the advances in research on gadolinium-containing carbon nanomaterials and their advantages over conventional gadolinium contrast agents. Due to their enhanced proton relaxivity, they can provide a reliable imaging contrast for cells, tissues or organs with much smaller doses than currently used in clinical practice, thus leading to reduced toxicity (as shown by cytotoxicity and biodistribution studies). Their active targeting capability allows for improved MRI of molecular or cellular targets, overcoming the limited labelling capability of available contrast agents (restricted to physiological irregularities during pathological conditions). Their potential of multifunctionality encompasses multimodal imaging and the combination of imaging and therapy.     

Evolution of the claustrum in Cnidaria: comparative anatomy reveals that it is exclusive to some species of Staurozoa and absent in Cubozoa

The claustrum in Cnidaria is a tissue in the gastrovascular cavity delimited by a central layer of mesoglea surrounded by gastrodermis (i.e., gastrodermis-mesoglea-gastrodermis), without communication with epidermis. By dividing the gastrovascular cavity, the four claustra provide an additional level of complexity. The presence of claustra in Cubozoa and Staurozoa has been used as evidence supporting a close relationship between these two cnidarian classes. However, the detailed anatomy of the claustrum has never been comparatively analyzed, rendering the evolution of this character among Cnidaria and its homology in Staurozoa and Cubozoa uncertain. This study provides a comparative investigation of the internal anatomy of the claustrum in Staurozoa and Cubozoa, addressing its evolutionary history based on recent phylogenetic hypotheses for Cnidaria. We conclude that the claustrum is a character exclusive to some species of Staurozoa, with a homoplastic evolution in the class, and that the structure called the “claustrum” in Cubozoa corresponds to the valve of gastric ostium, a structure at the base of the manubrium, which is also present in Staurozoa with and without claustrum. Thus, the claustrum cannot be a synapomorphy of a hypothetical clade uniting Staurozoa and Cubozoa, nor can its hypothetical presence in enigmatic fossils be used to support cubozoan affinities.     

Fatty acid profiles of the topmouth culter (Culter alburnus Basilewsky, 1855) from five lakes located at different latitudes in China

The topmouth culter (Culter alburnus Basilewsky, 1855) is a freshwater carnivorous fish of high nutritional value and widely distributed in lakes and reservoirs of China. In order to evaluate the fatty acid profiles in relation to habitat characteristics and to elucidate the main composition predictors, C. alburnus were collected from five lakes (Xingkai‐ LXk, Hongze‐ LHz, Kuilei‐ LKl, Chidong‐ LCd and Dongting‐ LDt) in China, ranging in latitudes from 28°30′ to 44°44′N and altitudes from 4 to 64 m. Ten fish from each lake ranging in total lengths from 414.2 to 423.9 mm were sampled between May to June in 2013 and 2014. A total of 23 fatty acids were identified using gas chromatography. Fatty acids that predominated in muscle samples of the fish were 16:0, 18:0, 16:1n?7, 18:1n?9, 20:5n?3, 22:6n?3 and 20:4n?6. Important differences in the fatty acid profiles of fish from high and low latitude lakes were discernible in that the two high latitudinal lakes (LXk and LHz) had significantly higher (p < .05) ratios of n?3 to n?6 fatty acids and levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), than those fishes in the low latitudinal lakes (LKl, LCd and LDt); these observations were confirmed by a principal component analysis. Therefore, it appears that multivariate analysis enables to distinguish fish from different lakes based on their characteristic fatty acid signatures; this would make a contribution to fish product traceability. The percentages of total saturated fatty acids increased significantly with increasing ambient temperature, and lakes at lower latitudes had higher levels of 17:0, 18:0, 18:2n?6, 20:3n?6 and lower levels of 16:1n?7 and 18:1n?9. Theoretical information criteria showed that the percentage of total saturated fatty acids varied more with temperature than with latitude and that this temperature‐dependent variation is nonlinear. Variation partitioning shows that most of the geographical variations are not only due to the nonlinear effects of temperature but also to the independent effects of the lake features. By contrast, no significant variations in fish lengths were found.     

The Bohemo‐Iberian regional chronostratigraphical scale for the Ordovician System and palaeontological correlations within South Gondwana

The Bohemo‐Iberian regional scale for South Gondwana, involving the ‘Mediterranean Province’, comprises five regional stages (Arenigian, Oretanian, Dobrotivian, Berounian and Kralodvorian) plus the global Tremadocian and Hirnantian. The predominance of shallow‐water taxa in those high‐latitude faunas imposes serious difficulties for correlating the regional succession with the formal global chronostratigraphy because of the almost total absence of the key graptolites and conodonts defining the base of the standard series, stages and stage slices. Instead, the abundant benthic faunas (trilobites, brachiopods, molluscs, echinoderms) of South Gondwanan origin largely dominated in the area from the middle Darriwilian to the late Katian. The poleward faunal migration of originally Avalonian, Baltic, Laurentian and even Asiatic taxa during the Boda Event of global warming progressively ends with the endemicity in the region, where the ensuing benthic assemblages were severely affected by the Hirnantian glaciation. The regional scale significantly improves the precision of correlations between Ordovician strata from SW and central Europe, North Africa and a large part of the Middle East. An updated record of palaeontological data from areas where Mediterranean faunas remain practically unknown, or are still poorly investigated, is also included. Palaeobiogeographical relationships based on the distribution of faunas across South Gondwana are suggested as an improvement for positioning many territories in modern palaeogeographical reconstructions and offer a constructive approach to problems related to the pre‐Variscan and pre‐Alpine orogenic puzzles.     

Function predicts the allometry of contest-related traits,but not sexual or male dimorphism in the amazonian tusked harvestman

Evolutionary Ecology - Animal contests involve threatening displays and physical coercion, which are respectively performed by threat devices used in mutual evaluation of size or strength, and...     

Allelic Diversity and Its Implications for the Rate of Adaptation

Genetic variation is usually estimated empirically from statistics based on population gene frequencies, but alternative statistics based on allelic diversity (number of allelic types) can provide complementary information. There is a lack of knowledge, however, on the evolutionary implications attached to allelic-diversity measures, particularly in structured populations. In this article we simulated multiple scenarios of single and structured populations in which a quantitative trait subject to stabilizing selection is adapted to different fitness optima. By forcing a global change in the optima we evaluated which diversity variables are more strongly correlated with both short- and long-term adaptation to the new optima. We found that quantitative genetic variance components for the trait and gene-frequency-diversity measures are generally more strongly correlated with short-term response to selection, whereas allelic-diversity measures are more correlated with long-term and total response to selection. Thus, allelic-diversity variables are better predictors of long-term adaptation than gene-frequency variables. This observation is also extended to unlinked neutral markers as a result of the information they convey on the demographic population history. Diffusion approximations for the allelic-diversity measures in a finite island model under the infinite-allele neutral mutation model are also provided.     

Embryo and Endosperm Phytochemicals from Polyembryonic Maize Kernels and Their Relationship with Seed Germination

Because of the growing worldwide demand for maize grain, new alternatives have been sought for breeding of this cereal, e.g., development of polyembryonic varieties, which agronomic performance could positively impact the grain yield per unit area, and nutritional quality. The objectives of this study were to (1) determine the phytochemicals present in the embryo and endosperm of grain from maize families with high, low, and null polyembryony frequency, which were planted at different locations, and (2) state the relationship between these compounds and seed germination. The extracted phytochemicals from corn were identified by HPLC-MS. The results showed that the genotype with the highest presence of phytochemicals was the brachytic population with high polyembryony called “BAP”, which also required less water during the germination process. The number of phytochemicals in both embryo and endosperm tissues was not related to the sowing location where they came from or the type of polyembryony. The number of different phytochemicals depended on the grain tissue from where they were extracted. The chemical compounds found in the different maize tissues were related to the development of the plant, either in roots or nibs because these are mainly associated with the lignin synthesis.     

Gibberellins signal nuclear import of PHOR1, a photoperiod-responsive protein with homology to Drosophila armadillo

S. tuberosum ssp. andigena potato plants require short days (SD) for tuberization. We have isolated PHOR1 (photoperiod-responsive 1), which shows upregulated expression in induced leaves (SD). PHOR1 encodes an arm repeat protein with homology to the Drosophila segment polarity protein armadillo. Antisense inhibition of PHOR1 produces a semidwarf phenotype similar to that of GA-deficient plants, and the antisense lines show reduced GA responsiveness combined with a higher endogenous GA content than wild-type plants. Feedback regulation of GA biosynthetic genes is also altered in these lines. Conversely, transgenic lines overexpressing PHOR1 show an enhanced response to GA. GA application induces rapid migration of PHOR1-GFP protein to the nucleus. Thus, PHOR1 appears to be a general component of GA signaling pathways that relocalizes to the nucleus in the presence of GA.