Exercise training attenuated the PKB and GSK-3 dephosphorylation in the myocardium of ZDF rats.

Cardiac dysfunction is a severe secondary effect of Type 2 diabetes. Recruitment of the protein kinase B/glycogen synthase kinase-3 pathway represents an integral event in glucose homeostasis, albeit its regulation in the diabetic heart remains undefined. Thus the following study tested the hypothesis that the regulation of protein kinase B/glycogen synthase kinase-3 was altered in the myocardium of the Zucker diabetic fatty rat. Second, exercise has been shown to improve glucose homeostasis, and, in this regard, the effect of swimming training on the regulation of protein kinase B/glycogen synthase kinase-3 in the diabetic rat heart was examined. In the sedentary Zucker diabetic fatty rats, glucose levels were elevated, and cardiac glycogen content increased, compared with wild type. A 13-wk swimming regimen significantly reduced plasma glucose levels and cardiac glycogen content and partially normalized protein kinase B-serine473, protein kinase B-threonine308, and glycogen synthase kinase-3alpha phosphorylation in Zucker diabetic fatty rats. In conclusion, hyperglycemia and increased cardiac glycogen content in the Zucker diabetic fatty rats were associated with dysregulation of protein kinase B/glycogen synthase kinase-3 phosphorylation. These anomalies in the Zucker diabetic fatty rat were partially normalized with swimming. These data support the premise that exercise training may protect the heart against the deleterious consequences of diabetes.     

HIERARCHICAL SPATIAL STRUCTURE AND DISCRIMINANT ANALYSIS OF GENETIC DIVERSITY IN THE RED ALGA MAZZAELLA LAMINARIOIDES (GIGARTINALES, RHODOPHYTA)

Our study of the genetic structure of Mazzaella laminarioides (Bory) Fredericq (Gigartinales) in the central Chilean region documented a high level of genetic diversity based on random amplified polymorphic DNA (RAPD) markers and indicated the occurrence of significant genetic structure at different spatial scales. A total of 288 haploid gametophytes was analyzed with 17 polymorphic RAPD bands, which produced 202 distinct multilocus genotypes. Within stands, mean gene diversity ranged from 0.210 to 0.249 and no significant linkage disequilibrium could be detected among pairs of alleles, revealing that recombination (sexual reproduction) regularly shuffles the genes at that scale. Analysis of molecular variance within stands (less than 30 m) showed that the structure was very low, only marginally significant, and did not increase with increasing hierarchical levels at this lowest spatial scale. In contrast, at a larger spatial scale (among stands, from 5 to 60 km), increasing geographical distance seemed to account for increasing isolation between populations even if natural barriers, such as sandy beaches or river estuaries, may play a role in such isolation. Moreover, the strong genetic differentiation occurring between locations separated by 60 km allowed the assignment of individuals to their original population through a canonical discriminant analysis. This approach further allowed the identification of potential recent migrants from one population to the other. Thus, in species like M. laminarioides for which the dominance of RAPD markers can be avoided by selecting haploid individuals, RAPD analysis appeared to be specially appropriate for the study of genetic differentiation.     

Apoptosis-resistant NS/0 E1B-19K myelomas exhibit increased viability and chimeric antibody productivity under cell cycle modulating conditions

Lymphoid cells expressing sufficient levels of Bcl-2 or E1B-19K are known to resist to induction of apoptosis in glutamine-free or nutrient-limited batch cultures. However, despite the increased viability and prolonged stationary phase achieved in batch culture, product yields are not necessarily improved. Here we have found that expression of E1B-19K in NS/0 myeloma cells cultivated in the presence of certain cell cycle modulators could result in a significant increase in MAb productivity as compared to untransfected control cells. The use of E1B-19K significantly enhanced cell survival in the presence of osmolytes (sorbitol, NaCl), DNA synthesis inhibitors (hydroxyurea, excess thymidine), and the cell culture additive OptiMAb™. E1B-19K myelomas cultivated in the presence of NaCl or OptiMAb™ accumulated in the G1 phase, while those arrested with excess thymidine were blocked in all phases. Interestingly, control NS/0 cells treated with these agents were found to die in a cell-cycle specific manner. Thus, while all G1 and most S phase cells quickly underwent apoptosis, G2/M cells remained alive and maintained MAb secretion for more than 10 days if supplied with adequate nutrients. For both control and E1B-19K cells, incubation with sorbitol or hydroxyurea was detrimental for MAb secretion, while addition of NaCl, excess thymidine and OptiMAb™ resulted in an increased specific MAb productivity as compared to the batch culture. However, this increase resulted in an improvement of final MAb yields only in the case of OptiMAb™. The extension of viability conferred by E1B-19K allowed to further improve the final MAb yield obtained using OptiMAb™ with a 3.3-fold increase for E1B-19K cells as compared to 1.8-fold for control NS/0 cells. This revised version was published online in August 2006 with corrections to the Cover Date.     

Crystal structure of Trypanosoma cruzi glyceraldehyde-3-phosphate dehydrogenase complexed with an analogue of 1,3-bisphospho-d-glyceric acid.

We report here the first crystal structure of a stable isosteric analogue of 1,3-bisphospho-d-glyceric acid (1,3-BPGA) bound to the catalytic domain of Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) in which the two phosphoryl moieties interact with Arg249. This complex possibly illustrates a step of the catalytic process by which Arg249 may induce compression of the product formed, allowing its expulsion from the active site. Structural modifications were introduced into this isosteric analogue and the respective inhibitory effects of the resulting diphosphorylated compounds on T. cruzi and Trypanosoma brucei gGAPDHs were investigated by enzymatic inhibition studies, fluorescence spectroscopy, site-directed mutagenesis, and molecular modelling. Despite the high homology between the two trypanomastid gGAPDHs (> 95%), we have identified specific interactions that could be used to design selective irreversible inhibitors against T. cruzi gGAPDH.     

Isolation-by-distance and male-biased dispersal at a fine spatial scale: a study of the common European adder (Vipera berus) in a rural landscape

Conservation Genetics - Human activities have a major impact on habitat connectivity and landscape structure. In this context, it is critical to better understand animal movements and gene flow to...     

Assessing community functional attributes during substrate colonization: a field experiment using stream insects

Initial colonists of empty habitats tend to differ from those arriving later in terms of species identity and traits. We evaluated the dynamics of the functional attributes in aquatic insect communities during a colonization experiment under natural conditions. We tested whether the late stages of colonization show higher functional richness, diversity and specialization than early successional stages. We used 60 artificial slate samplers that were removed after 1, 3, 5, 10, 15 and 30 days of colonization. We considered five traits (with a total of 17 trait categories): feeding habits, dispersal medium, body size, body shape and locomotion. With these traits, we computed a global specialization index at the community level. Large shredders with a cylindrical body shape and fly dispersal while flying were prominent in late colonization. In contrast, early colonists tended to have flattened body and to disperse through water. Functional diversity and functional richness significantly increased in late colonization, resulting in a more specialized community. Our results suggest that any interference during the processes involved in stream insect colonization can be reflected in the community through the decrease or even lack of functional attributes.

     

Streamlining CRISPR spacer-based bacterial host predictions to decipher the viral dark matter

Thousands of new phages have recently been discovered thanks to viral metagenomics. These phages are extremely diverse and their genome sequences often do not resemble any known phages. To appreciate their ecological impact, it is important to determine their bacterial hosts. CRISPR spacers can be used to predict hosts of unknown phages, as spacers represent biological records of past phage–bacteria interactions. However, no guidelines have been established to standardize host prediction based on CRISPR spacers. Additionally, there are no tools that use spacers to perform host predictions on large viral datasets. Here, we developed a set of tools that includes all the necessary steps for predicting the hosts of uncharacterized phages. We created a database of >11 million spacers and a program to execute host predictions on large viral datasets. Our host prediction approach uses biological criteria inspired by how CRISPR–Cas naturally work as adaptive immune systems, which make the results easy to interpret. We evaluated the performance using 9484 phages with known hosts and obtained a recall of 49% and a precision of 69%. We also found that this host prediction method yielded higher performance for phages that infect gut-associated bacteria, suggesting it is well suited for gut-virome characterization.