Soybean ferritin: isolation, characterization, and free radical generation

The main aim of this work was to assess the multi-task role of ferritin(Ft)in the oxidative metabolism of soybean(Glycine max).Soybean seeds incubated for 24 h yielded 41 ± 5 μg Ft/g fresh weight.The rate of in vitro incorporation of iron(Fe)into Ft was tested by supplementing the reaction medium with physiological Fe chelators.The control rate,observed in the presence of 100 μM Fe,was not significantly different from the values observed in the presence of 100 μM Fe-his.However,it was significantly higher in the presence of 100 μM Fe-citrate(approximately 4.5-fold)or of 100 μM Fe-ATP(approximately 14-fold).Moreover,a substantial decrease in the Trp-dependent fluorescence of the Ft protein was determined during Fe uptake from Fe-citrate,as compared with the control.On the other hand,Ft addition to homogenates from soybean embryonic axes reduced endogenously generated ascorbyl radical,according to its capacity for Fe uptake.The data presented here suggest that Ft could be involved in the generation of free radicals,such as hydroxyl radical,by Fe-catalyzed reactions.Moreover,the scavenging of these radicals by Ft itself could then lead to protein damage.However,Ft could also prevent cellular damage by the uptake of catalytically active Fe.     

Population structure of North Atlantic and North Pacific sei whales (<Emphasis Type="Italic">Balaenoptera borealis</Emphasis>) inferred from mitochondrial control region DNA sequences and microsatellite genotypes

Currently, three stocks of sei whales (Balaenoptera borealis) are defined in the North Atlantic; the Nova Scotian, Iceland-Denmark Strait and Eastern North Atlantic stocks, which are mainly based upon historical catch and sighting data. We analyzed mitochondrial control region DNA (mtDNA) sequences and genotypes from 7 to 11 microsatellite loci in 87 samples from three sites in the North Atlantic; Iceland, the Gulf of Maine and the Azores, and compared against the North Pacific using 489 previously published samples. No statistically significant deviations from homogeneity were detected among the North Atlantic samples at mtDNA or microsatellite loci. The genealogy estimated from the mtDNA sequences revealed a clear division of the haplotypes into a North Atlantic and a North Pacific clade, with the exception of one haplotype detected in a single sample from the Azores, which was included in the North Pacific clade. Significant genetic divergence between the North Atlantic and North Pacific Oceans was detected (mtDNA Φ_ST?=?0.72, microsatellite Weir and Cockerham’s ? = 0.20; p?<?0.001). The coalescent-based estimate of the population divergence time between the North Atlantic and North Pacific populations from the sequence variation among the mtDNA sequences was at 163,000 years ago. However, the inference was limited by an absence of samples from the Southern Hemisphere and uncertainty regarding mutation rates and generation times. The estimates of inter-oceanic migration rates were low (Nm at 0.007 into the North Pacific and at 0.248 in the opposite direction). Although estimates of genetic divergence among the current North Atlantic stocks were low and consistent with the extensive range of movement observed in satellite tagged sei whales, the high uncertainty of the genetic divergence estimates precludes rejection of multiple stocks in the North Atlantic.     

Effect of High-Fidelity Simulation on Medical Students’ Knowledge about Advanced Life Support: A Randomized Study

High-fidelity simulation (HFS) is a learning method which has proven effective in medical education for technical and non-technical skills. However, its effectiveness for knowledge acquisition is less validated. We performed a randomized study with the primary aim of investigating whether HFS, in association with frontal lessons, would improve knowledge about advanced life support (ALS), in comparison to frontal lessons only among medical students. The secondary aims were to evaluate the effect of HFS on knowledge acquisition of different sections of ALS and personal knowledge perception. Participants answered a pre-test questionnaire consisting of a subjective (evaluating personal perception of knowledge) and an objective section (measuring level of knowledge) containing 100 questions about algorithms, technical skills, team working/early warning scores/communication strategies according to ALS guidelines. All students participated in 3 frontal lessons before being randomized in group S, undergoing a HFS session, and group C, receiving no further interventions. After 10 days from the end of each intervention, both groups answered a questionnaire (post-test) with the same subjective section but a different objective one. The overall number of correct answers of the post-test was significantly higher in group S (mean 74.1, SD 11.2) than in group C (mean 65.5, SD 14.3), p = 0.0017, 95% C.I. 3.34 – 13.9. A significantly higher number of correct answers was reported in group S than in group C for questions investigating knowledge of algorithms (p = 0.0001; 95% C.I 2.22–5.99) and team working/early warning scores/communication strategies (p = 0.0060; 95% C.I 1.13–6.53). Students in group S showed a significantly higher score in the post-test subjective section (p = 0.0074). A lower proportion of students in group S confirmed their perception of knowledge compared to group C (p = 0.0079). HFS showed a beneficial effect on knowledge of ALS among medical students, especially for notions of algorithms and team working/early warning scores/communication.     

Structural role of PufX in the dimerization of the photosynthetic core complex of Rhodobacter sphaeroides

Monomeric and dimeric PufX-containing core complexes have been purified from membranes of wild-type Rhodobacter sphaeroides. Reconstitution of both samples by detergent removal in the presence of lipids leads to the formation of two-dimensional crystals constituted of dimeric core complexes. Two-dimensional crystals were further analyzed by cryoelectron microscopy and atomic force microscopy. A projection map at 26-A resolution reveals that core complexes assemble in an "S"-shaped dimeric complex. Each core complex is composed of one reaction center, 12 light-harvesting 1 alpha/beta-heterodimers, and one PufX protein. The light-harvesting 1 assemblies are open with a gap of density of approximately 30-A width and surround oriented reaction centers. A maximum density is found at the dimer junction. Based on the projection map, a model is proposed, in which the two PufX proteins are located at the dimer junction, consistent with the finding of dimerization of monomeric core complexes upon reconstitution. This localization of PufX in the core complex implies that PufX is the structural key for the dimer complex formation rather than a channel-forming protein for the exchange of ubiquinone/ubiquinol between the reaction center and the cytochrome bc1 complex.     

Development of an assay to screen for inhibitors of tau phosphorylation by cdk5

A high-throughput assay for tau phosphorylation by cdk5/p25 is described. Full-length recombinant tau was used as a substrate in the presence of saturating adenosine triphosphate (ATP). Using PHF-1, an antibody directed specifically against 2 tau phosphorylation epitopes (serine 396 and serine 404), an enzyme-linked immunosorbent assay (ELISA)-based colorimetric assay was formatted in 384-well plates. The assay was validated by measuring kinetic parameters for cdk5/p25 catalysis and known inhibitors. Rate constants for the site-specific phosphorylations at the PHF-1 epitopes were determined and suggested preferential phosphorylation at these sites. The performance of this assay in a high-throughput format was demonstrated and used to identify inhibitors of tau phosphorylation at specific epitopes phosphorylated by cdk5/p25.