Analysis of oxidative stress-induced protein carbonylation using fluorescent hydrazides

Protein carbonyl detection has been commonly used to analyze the degree of damage to proteins under oxidative stress conditions. Most laboratories rely on derivatization of carbonyl groups with dinitrophenylhydrazine followed by Western blot analysis using antibodies against the dinitrophenyl moiety. This paper describes a protein carbonyl detection method based on fluorescent Bodipy, Cy3 and Cy5 hydrazides. Using this approach, Western blot and immunodetection are no longer needed, shortening the procedure and increasing accuracy. Combination of Cy3 and Cy5 hydrazides allows multiplexing analyses in a single two-dimensional gel. Derivatization with Bodipy hydrazide allows easy matching of the spots of interest and those obtained by general fluorescent protein staining methods, which facilitates excising target proteins from the gels and identifying them. This method is effective for detecting protein carbonylation in samples of proteins submitted to metal-catalyzed oxidation "in vitro" and assessing the effect of hydrogen peroxide and chronological aging on protein oxidative damage in yeast cells.     

Detection of quantitative trait loci for resistance to gastrointestinal nematode infections in Creole goats

This study aimed to identify regions of the genome affecting resistance to gastrointestinal nematodes in a Creole goat population naturally exposed to a mixed nematode infection (Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum columbianum) by grazing on irrigated pasture. A genome‐wide quantitative trait loci (QTL) scan was performed on 383 offspring from 12 half‐sib families. A total of 101 microsatellite markers were genotyped. Traits analysed were faecal egg count (FEC), packed cell volume (PCV), eosinophil count and bodyweight (BW) at 7 and 11 months of age. Levels of activity of immunoglobulin A (IgA) and activity of immunoglobulin E (IgE) anti‐Haemonchus contortus L3 crude extracts and adult excretion/secretion products (ESPs) were also analysed. Using interval mapping, this study identified 13 QTL for parasite resistance. Two QTL linked with FEC were found on chromosomes 22 and 26. Three QTL were detected on chromosomes 7, 8 and 14 for eosinophil counts. Three QTL linked with PCV were identified on chromosomes 5, 9 and 21. A QTL for BW at 7 months of age was found on chromosome 6. Lastly, two QTL detected on chromosomes 3 and 10 were associated with IgE anti‐L3, and IgE anti‐ESP was linked with two QTL on chromosomes 1 and 26. This study is the first to have identified regions of the genome linked with nematode resistance in a goat population using a genome scan. These results provide useful tools for the understanding of parasite resistance in small ruminants.     

How does hydroxyl introduction influence the double helical structure: the stabilization of an altritol nucleic acid:ribonucleic acid duplex

Altritol nucleic acids (ANAs) are a promising new tool in the development of artificial small interfering ribonucleic acids (siRNAs) for therapeutical applications. To mimic the siRNA:messenger RNA (mRNA) interactions, the crystal structure of the ANA:RNA construct a(CCGUAAUGCC-P):r(GGCAUUACGG) was determined to 1.96?? resolution which revealed the hybrid to form an A-type helix. As this A-form is a major requirement in the RNAi process, this crystal structure confirms the potential of altritol-modified siRNAs. Moreover, in the ANA strands, a new type of intrastrand interactions was found between the O2' hydroxyl group of one residue and the sugar ring O4' atom of the next residue. These interactions were further investigated by quantum chemical methods. Besides hydration effects, these intrastrand hydrogen bonds may also contribute to the stability of ANA:RNA duplexes.     

Tassements vertébraux chez le petit enfant : présentation d’un cas et discussion étiologique

Vertebral compression fractures among children are less common and described than among adults. We report a case of a 3-year-old boy presenting with low back pain. The bone scan revealed multiple vertebral compression fractures. This case leads us to discuss the main etiologies of vertebral compression fractures in pediatric population: traumatic causes, secondary osteoporosis and its many underlying etiologies, and primary osteoporosis (mainly osteogenesis imperfecta and idiopathic juvenile osteoporosis).     

Isolation and cDNA cloning of an Em-like protein from mung bean (Vigna radiata) axes

Until now no 'early-methionine-labelled' (Em) proteins have been reported in the Fabaceae. To check whether a previously isolated low-molecular mass albumin from dry mung bean embryonic axes possibly corresponded to an Em-like protein, the protein was purified, sequenced and its cDNA clone isolated and characterized. N-terminal sequencing of cyanogen bromide cleavage products of the protein revealed homology with previously described Em-like proteins from other species. Analysis of cDNA clones encoding the mung bean Em protein revealed the presence of two classes of Em proteins and confirmed their homology to the previously characterized Em-like proteins. In vivo labelling and northern blot analysis further demonstrated that the mung bean protein is synthesized during early germination of the axes and that abscisic acid (ABA) extends its synthesis. It appears, therefore, that legumes also contain maturation-specific, ABA-responsive Em-like proteins.     

Abundance and Size of Gulf Shrimp in Louisiana's Coastal Estuaries following the Deepwater Horizon Oil Spill

The Deepwater Horizon oil spill impacted Louisiana''s coastal estuaries physically, chemically, and biologically. To better understand the ecological consequences of this oil spill on Louisiana estuaries, we compared the abundance and size of two Gulf shrimp species (Farfantepeneus aztecus and Litopeneus setiferus) in heavily affected and relatively unaffected estuaries, before and after the oil spill. Two datasets were used to conduct this study: data on shrimp abundance and size before the spill were available from Louisiana Department of Wildlife and Fisheries (LDWF). Data on shrimp abundance and size from after the spill were independently collected by the authors and by LDWF. Using a Before-After-Control-Impact with Paired sampling (BACIP) design with monthly samples of two selected basins, we found brown shrimp to become more abundant and the mean size of white shrimp to become smaller. Using a BACIP with data on successive shrimp year-classes of multiple basins, we found both species to become more abundant in basins that were affected by the spill, while mean shrimp size either not change after the spill, or increased in both affected and unaffected basins. We conclude that following the oil spill abundances of both species increased within affected estuaries, whereas mean size may have been unaffected. We propose two factors that may have caused these results: 1) exposure to polycyclic aromatic hydrocarbons (PAHs) may have reduced the growth rate of shrimp, resulting in a delayed movement of shrimp to offshore habitats, and an increase of within-estuary shrimp abundance, and 2) fishing closures established immediately after the spill, may have resulted in decreased fishing effort and an increase in shrimp abundance. This study accentuates the complexities in determining ecological effects of oil spills, and the need of studies on the organismal level to reveal cause-and-effect relationships of such events.