Genetic Variability and Linkage Disequilibrium Patterns in the Bovine <Emphasis Type="Italic">DNAJA1</Emphasis> Gene

Correlation between expression level of the bovine DNAJA1 gene and meat tenderness was recently found in Charolais longissimus thoracis muscle samples, suggesting that this gene could play an important role in meat tenderness. Here, we report the validation of polymorphisms within the bovine DNAJA1 gene, and the haplotype variability and extent of linkage disequilibrium in the three main French beef breeds (Blonde d’Aquitaine, Charolais, Limousin). Genotyping 18 putative SNPs revealed that 16 SNPs were polymorphic within the breeds tested. Two SNPs were removed from further analyses as one SNP had a low genotyping call rate, while the other SNP was not in Hardy–Weinberg equilibrium. The degree of heterozygosity observed for the remaining 14 SNPs varied between breeds, with Charolais being the breed with the highest genetic variation and Blonde d’Aquitaine the lowest. Linkage disequilibrium and haplotype structure of DNAJA1 were different between breeds. Eighteen different haplotypes, including three shared by all breeds, were discovered, and two to three tag SNPs (depending on the breed) are sufficient to capture all the genetic variability seen in these haplotypes. The results of this study will facilitate the design of optimal future association studies evaluating the role of the DNAJA1 gene in meat tenderness.     

High-level swimmers' kinetic efficiency during the underwater phase of a grab start

The purpose of the present work was to study swimmers' efficiency during the underwater phase of the grab start. Eight high-level swimmers participated in this study. They performed two types of start: a regular grab start (with underwater leg propulsion after the glide) and a grab start with no underwater movement (swimmers had to remain in a streamlined position). Four cameras filmed the entire underwater phase of all starts. Nine anatomic landmarks were identified on the swimmers' bodies and their positions were calculated using a modified double plan DLT technique. From these positions and Dempster's anthropometric data, the center of mass position and velocity were also determined. Kinetic energies were also calculated. This velocity and kinetic energies for the two types of start were compared. Swimmers began underwater leg propulsion 1.69 m too soon. The global and internal energies were significantly higher for the start with underwater leg propulsion. Nevertheless, swimmers' velocities were equivalent for both starts. These results suggest that the swimmers did not use the underwater phase of the start efficiently: By kicking too soon, they did not succeed in producing higher velocities and thus wasted energy.     

Phytostabilisation of copper-contaminated soil in Katanga: an experiment with three native grasses and two amendments

This study evaluates the feasibility of using the grass species Rendlia altera, Monocymbium ceresiiforme, Cynodon dactylon, and amendments (compost and lime) for the phytostabilisation of soils contaminated by Cu in the province of Katanga (Democratic Republic of Congo). Species were grown on control and Cu-contaminated plots (artificially contaminated with 2,500 mg kg(-1) Cu) unamended (NA), amended with 4.5 kg compost m(-2) or 0.2 kg lime m(-2). R. altera was also grown on contaminated plots amended with 22.5 kg compost m(-2) or 1 kg lime m(-2). Plant survival, growth, and reproduction were monitored for two years. Cu-concentration in leaves of R. altera and M. ceresiiforme were analysed. pH and extractable Cu (0.01 M CaCl2) in soil were analysed in April 2007 and 2008. Results showed that R. altera seems to be the best candidate because of its highest survival on NA, followed by M. ceresiiforme, while liming was necessary to ensure survival of C. dactylon. Lime increased plant reproduction and reduced Cu accumulation in leaves compared to compost. However, higher survival and number of spikes of R. altera obtained in experiment 2 with 22.5 kg compost m(-2) suggest that lime x compost interactions should be investigated in further studies.     

Assessment of a Syndromic Surveillance System Based on Morbidity Data: Results from the Oscour? Network during a Heat Wave

Background

Syndromic surveillance systems have been developed in recent years and are now increasingly used by stakeholders to quickly answer questions and make important decisions. It is therefore essential to evaluate the quality and utility of such systems. This study was designed to assess a syndromic surveillance system based on emergency departments'' (ED) morbidity rates related to the health effects of heat waves. This study uses data collected during the 2006 heat wave in France.

Methods

Data recorded from 15 EDs in the Ile-de-France (Paris and surrounding area) from June to August, 2006, were transmitted daily via the Internet to the French Institute for Public Health Surveillance. Items collected included diagnosis (ICD10), outcome, and age. Several aspects of the system have been evaluated (data quality, cost, flexibility, stability, and performance). Periods of heat wave are considered the most suitable time to evaluate the system.

Results

Data quality did not vary significantly during the period. Age, gender and outcome were completed in a comprehensive manner. Diagnoses were missing or uninformative for 37.5% of patients. Stability was recorded as being 99.49% for the period overall. The average cost per day over the study period was estimated to be €287. Diagnoses of hyperthermia, malaise, dehydration, hyponatremia were correlated with increased temperatures. Malaise was most sensitive in younger and elderly adults but also the less specific. However, overall syndrome groups were more sensitive with comparable specificity than individual diagnoses.

Conclusion

This system satisfactorily detected the health impact of hot days (observed values were higher than expected on more than 90% of days on which a heat alert was issued). Our findings should reassure stakeholders about the reliability of health impact assessments during or following such an event. These evaluations are essential to establish the validity of the results of syndromic surveillance systems.     

Chloroplast DNA Polymorphism and Phylogeography of a Central African Tree Species Widespread in Mature Rainforests: Greenwayodendron suaveolens (Annonaceae)

Geographic patterns of genetic variation at chloroplast markers have been successfully used to address the phylogeography and the demographic history of many plant species. Very few studies have however been conducted in important tropical centers of plant biodiversity like the African rainforests. The phylogeography of a tree species widespread in Central African mature forests, Greenwayodendron suaveolens subsp. suaveolens (Annonaceae), was investigated in the Lower Guinea phytogeographic domain (essentially Gabon and Cameroon) by sequencing an intergenic spacer of the chloroplast genome (trnC-petN1R). A total of 11 polymorphic sites, including nine single nucleotide polymorphisms (SNPs), two insertions/deletions and two inversions, defined 12 haplotypes. The taxon is represented by two sympatric varieties (var. suaveolens and var. gabonica) that carried distinct and relatively divergent haplotypes. These varieties, also well distinguishable morphologically, might therefore represent true biological species. The variety suaveolens, more common and more widespread than the variety gabonica, was represented by ten haplotypes. This taxon showed a weak but statistically significant phylogeographic structure, indicating that two sets of related haplotypes essentially occurred respectively in the northern and the southern hemispheres. These results suggest that the distribution of Greenwayodendron suaveolens subsp. suaveolens, which is currently continuous in the Lower Guinea domain, might have been more fragmented in the past, possibly in relation with Pleistocene forest fragmentation.     

Characterisation of the expression of NMDA receptors in human astrocytes

Astrocytes have long been perceived only as structural and supporting cells within the central nervous system (CNS). However, the discovery that these glial cells may potentially express receptors capable of responding to endogenous neurotransmitters has resulted in the need to reassess astrocytic physiology. The aim of the current study was to characterise the expression of NMDA receptors (NMDARs) in primary human astrocytes, and investigate their response to physiological and excitotoxic concentrations of the known endogenous NMDAR agonists, glutamate and quinolinic acid (QUIN). Primary cultures of human astrocytes were used to examine expression of these receptors at the mRNA level using RT-PCR and qPCR, and at the protein level using immunocytochemistry. The functionality role of the receptors was assessed using intracellular calcium influx experiments and measuring extracellular lactate dehydrogenase (LDH) activity in primary cultures of human astrocytes treated with glutamate and QUIN. We found that all seven currently known NMDAR subunits (NR1, NR2A, NR2B, NR2C, NR2D, NR3A and NR3B) are expressed in astrocytes, but at different levels. Calcium influx studies revealed that both glutamate and QUIN could activate astrocytic NMDARs, which stimulates Ca2+ influx into the cell and can result in dysfunction and death of astrocytes. Our data also show that the NMDAR ion channel blockers, MK801, and memantine can attenuate glutamate and QUIN mediated cell excitotoxicity. This suggests that the mechanism of glutamate and QUIN gliotoxicity is at least partially mediated by excessive stimulation of NMDARs. The present study is the first to provide definitive evidence for the existence of functional NMDAR expression in human primary astrocytes. This discovery has significant implications for redefining the cellular interaction between glia and neurons in both physiological processes and pathological conditions.     

Agrobacterium tumefaciens type II NADH dehydrogenase. Characterization and interactions with bacterial and thylakoid membranes

Type II NADH dehydrogenases (NDH-2) are monomeric enzymes that catalyse quinone reduction and allow electrons to enter the respiratory chain in different organisms including higher plant mitochondria, bacteria and yeasts. In this study, an Agrobacterium tumefaciens gene encoding a putative alternative NADH dehydrogenase (AtuNDH-2) was isolated and expressed in Escherichia coli as a (His)6-tagged protein. The purified 46 kDa protein contains FAD as a prosthetic group and oxidizes both NADH and NADPH with similar Vmax values, but with a much higher affinity for NADH than for NADPH. AtuNDH-2 complements the growth (on a minimal medium) of an E. coli mutant strain deficient in both NDH-1 and NDH-2, and is shown to supply electrons to the respiratory chain when incubated with bacterial membranes prepared from this mutant. By measuring photosystem II chlorophyll fluorescence on thylakoid membranes prepared from the green alga Chlamydomonas reinhardtii, we show that AtuNDH-2 is able to stimulate NADH-dependent reduction of the plastoquinone pool. We discuss the possibility of using heterologous expression of NDH-2 enzymes to improve nonphotochemical reduction of plastoquinones and H2 production in C. reinhardtii.     

Highly variable rates of genome rearrangements between hemiascomycetous yeast lineages

Hemiascomycete yeasts cover an evolutionary span comparable to that of the entire phylum of chordates. Since this group currently contains the largest number of complete genome sequences it presents unique opportunities to understand the evolution of genome organization in eukaryotes. We inferred rates of genome instability on all branches of a phylogenetic tree for 11 species and calculated species-specific rates of genome rearrangements. We characterized all inversion events that occurred within synteny blocks between six representatives of the different lineages. We show that the rates of macro- and microrearrangements of gene order are correlated within individual lineages but are highly variable across different lineages. The most unstable genomes correspond to the pathogenic yeasts Candida albicans and Candida glabrata. Chromosomal maps have been intensively shuffled by numerous interchromosomal rearrangements, even between species that have retained a very high physical fraction of their genomes within small synteny blocks. Despite this intensive reshuffling of gene positions, essential genes, which cluster in low recombination regions in the genome of Saccharomyces cerevisiae, tend to remain syntenic during evolution. This work reveals that the high plasticity of eukaryotic genomes results from rearrangement rates that vary between lineages but also at different evolutionary times of a given lineage.     

An integrated metabonomic approach to describe temporal metabolic disregulation induced in the rat by the model hepatotoxin allyl formate

The time-related metabolic events in rat liver, plasma, and urine following hepatotoxic insult with allyl formate (75 mg/kg) were studied using a combination of high-resolution liquid state and magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopic methods together with pattern recognition analysis. The metabonomics results were compared with the results of conventional plasma chemistry and histopathological assessments of liver damage. Various degrees of liver damage were observed in different animals, and this variation was reflected in all of the analyses. Furthermore, each analysis revealed a high degree of functional and structural recovery by the end of the study. The allyl formate-induced changes included hepatocellular necrosis, hepatic lipidosis, decreased liver glycogen and glucose, decreased plasma lipids, increased plasma creatine and tyrosine, increased urinary taurine and creatine, and decreased urinary TCA cycle intermediates. The observed reductions in hepatic glycogen and glucose suggest increased glucose utilization and are consistent with the expected depletion of hepatic ATP following mitochondrial impairment, assuming that there is a consequent increase in energy production from glycolysis. The increase in plasma tyrosine is consistent with impaired protein synthesis, a known consequence of ATP depletion. Partial least squares-based cross-correlation of the variation in the liver and plasma NMR profiles indicated that the allyl formate-induced increase in liver lipids correlated with the decrease in plasma lipids. This suggests disruption in lipid transport from the liver to plasma, which could arise through impaired apolipoprotein synthesis, as with ethionine.     

A new two-color Fab labeling method for autoantigen protein microarrays

Antigen microarrays hold great promise for profiling the humoral immune response in the settings of autoimmunity, allergy and cancer. This approach involves immobilizing antigens on a slide surface and then exposing the array to biological fluids containing immunoglobulins. Although these arrays have proven extremely useful as research tools, they suffer from several sources of variability. To address these issues, we have developed a new two-color Fab labeling method that allows two samples to be applied simultaneously to the same array. This straightforward labeling approach improves reproducibility and reliably detects changes in autoantibody concentrations. Using this technique we profiled serum from a mouse model of systemic lupus erythematosus (SLE) and detected both expected and previously unrecognized reactivities. The improved labeling and detection method described here overcomes several problems that have hindered antigen microarrays and should facilitate translation to the clinical setting.