Effects of genotype and environment on free amino acid levels in wheat grain: implications for acrylamide formation during processing

Acrylamide forms from free asparagine and reducing sugars during cooking, with asparagine concentration being the key parameter determining the formation in foods produced from wheat flour. In this study free amino acid concentrations were measured in the grain of varieties Spark and Rialto and four doubled haploid lines from a Spark x Rialto mapping population. The parental and doubled haploid lines had differing levels of total free amino acids and free asparagine in the grain, with one line consistently being lower than either parent for both of these factors. Sulfur deprivation led to huge increases in the concentrations of free asparagine and glutamine, and canonical variate analysis showed clear separation of the grain samples as a result of treatment (environment, E) and genotype (G) and provided evidence of G x E interactions. Low grain sulfur and high free asparagine concentration were closely associated with increased risk of acrylamide formation. G, E, and G x E effects were also evident in grain from six varieties of wheat grown at field locations around the United Kingdom in 2006 and 2007. The data indicate that progress in reducing the risk of acrylamide formation in processed wheat products could be made immediately through the selection and cultivation of low grain asparagme varieties and that further genetically driven improvements should be achievable. However, genotypes that are selected should also be tested under a range of environmental conditions.

Foraminifera and microfossil occurence in Late Jurassic sediments of the North and South Atlantic

Biostratigraphical, taxonomical, and palaeocological results were obtained from Oxfordian to Tithonian foraminifers of the Northern and Southern Atlantic Ocean boreholes of the DSDP Legs 1, 11, 36, 41, 44, 50, and 79. An oversight on the cored Jurassic sections of the DSDP Legs 79 and the corresponding foraminiferal descriptions are given. The reddish brown, clayey and carbonaceous Cat Gap Formation (Oxfordian to Tithonian) of the Northern Atlantic Ocean, rich in radiolarians, yields less or more uniform, in most cases allochthonous foraminiferal faunas of Central European shelf character. No Callovian and Upper Tithonian foraminiferaI zones can be established. The zone of Pseudomarssonella durnortieri covers the Oxfordian/Kimmeridgian, the zone of Neobulimina atlantica the Kimmeridgian/Lower Tithonian interval. Characteristic foraminiferal faunas are missing since the Upper Tithonian to Valanginian for reason of a widely distributed regression which caused hiatuses observed all over the Northern Atlantic Ocean and in parts of Europe. The Upper Jurassic cannot be subdivided into single stages by foraminiferal biostratigraphy alone. The fovaminiferal zones established by Moullad (1984) covering a Callovian-Tithonian interval may be of some local importance in the Tethyan realm: It has too long-ranging foraminiferal species to be used as index marker in the word-wide DSDP boreholes. Some taxonomical confusion is caused because in former publications some foraminiferal species have got different names both in the Jurassic and Cretaceous. The foraminiferal biostratigraphy of drilled sections from DSDP boreholes is restricted by the drilling technique and for palaeo-oceanographical, biological, and geological reasons. Foraminiferal faunas from the DSDP originally described as ,,bathyal, or ,,abyssal,, have to be derived from shallower water. This contrasts the palaeo-water depths of 3000-4000 m which result from sedimentological and palaeo-geographical investigations.

Marine diatoms in deep sea sediments

Long-term evolution is thought to take opportunities that arise as a consequence of mass extinction (as argued, for example, by Gould, 2002) and the following biotic recovery, but there is absolutely no evidence for this being the case. However, our study shows that eutrophication by oceanic mixing also played a part in the enhancement of several evolutionary events amongst marine organisms, and these results could indicate that the rates of oceanic biodiversification may be slowed if upwelling becomes weakened by future global warming. This paper defines three distinct evolutionary events of resting spores of the marine diatom genus Chaetoceros, to reconstruct past upwelling through the analysis of several DSDP, ODP and land-based successions from the North, South and equatorial Pacific as well as the Atlantic Ocean during the past 40 million years. The Atlantic Chaetoceros Explosion (ACE) event occurred across the E/O boundary in the North Atlantic, and is characterized by resting spore diversification that occurred as a consequence of the onset of upwelling following changes in thermohaline circulation through global cooling in the early Oligocene. Pacific Chaetoceros Explosion events-1 and -2 (PACE-1 and PACE-2) are characterized by relatively higher occurrences of iron input following the Himalayan uplift and aridification at 8.5 Ma and ca. 2.5 Ma in the North Pacific region. These events not only enhanced the diversification and increased abundance of primary producers, including that of Chaetoceros, other diatoms and seaweeds, but also stimulated the evolution of zooplankton and larger predators, such as copepods and marine mammals, which ate these phytoplankton and plants. Current thinking suggests new evolutionary niches open up after a mass extinction, but our study finds that eutrophication can also stimulate evolutionary diversification. Moreover, in the opposite fashion, our results show that as thermohaline circulation abates, global warming progresses and the ocean surface becomes warmer, many marine organisms will be affected by the environmental degradation.

G-quadruplex Formation Enhances Splicing Efficiency Of Pax9 Intron 1.

G-quadruplexes are secondary structures present in DNA and RNA molecules, which are formed by stacking of G-quartets (i.e., interaction of four guanines (G-tracts) bounded by Hoogsteen hydrogen bonding). Human PAX9 intron 1 has a putative G-quadruplex-forming region located near exon 1, which is present in all known sequenced placental mammals. Using circular dichroism (CD) analysis and CD melting, we showed that these sequences are able to form highly stable quadruplex structures. Due to the proximity of the quadruplex structure to exon-intron boundary, we used a validated double-reporter splicing assay and qPCR to analyze its role on splicing efficiency. The human quadruplex was shown to have a key role on splicing efficiency of PAX9 intron 1, as a mutation that abolished quadruplex formation decreased dramatically the splicing efficiency of human PAX9 intron 1. The less stable, rat quadruplex had a less efficient splicing when compared to human sequences. Additionally, the treatment with 360A, a strong ligand that stabilizes quadruplex structures, further increased splicing efficiency of human PAX9 intron 1. Altogether, these results provide evidences that G-quadruplex structures are involved in splicing efficiency of PAX9 intron 1.

Two new rodent genera from the Oligocene White River formation (family Heteromyidae) / William G. Reeder --

v.10:no.35(1960)

Étude moléculaire de la formation de complexes protéiques impliqués dans la signalisation des récepteurs couplés aux protéines G

La communication cellulaire est un phénomène important pour le maintien de l’homéostasie des cellules. Au court des dernières années, cette sphère de recherche sur la signalisation cellulaire a connue des avancées importantes au niveau de l’identification des acteurs principaux impliqués dans la reconnaissance extracellulaire des signaux, ainsi que la compréhension des voies de signalisation engagées par les cellules pour répondre aux facteurs extracellulaires. Malgré ces nouvelles informations, les diverses interrelations moléculaires entre les acteurs ainsi que les voies de signalisation cellulaire, demeurent mal comprises. Le transfert d’énergie de résonance de bioluminescence (BRET) permet la mesure d’interactions protéiques et peut être utilisé dans deux configurations, le BRET480-YFP (connu aussi comme le BRET1) et le BRET400-GFP (connu aussi en tant que BRET2). Suite à l’oxydation de son substrat, la luciférase de renilla peut transférer son énergie à une protéine fluorescente, uniquement si elles sont à proximité l’une de l’autre (≤100Å). La combinaison dans un seul essai des BRET480-YFP et BRET400-GFP, a permis de suivre trois paires d’interactions, sur une même population cellulaire. Par contre, l’utilisation de deux substrats pour la réaction de bioluminescence rend impossible la mesure simultanée des différents signaux de BRET, pour ce trois nouvelles configurations de BRET ont été mises au point en utilisant des nouvelles protéines fluorescentes. Ainsi deux des nouvelles couleurs de BRET ayant des émissions résolues, le BRET400-BFP et le BRET400mAmetrine ont pu être combinées pour mesurer l’engagement par un RCPG d’une protéine G, ainsi que l’accumulation du second messager. La combinaison de ces BRET a également permis de révéler la formation d’un complexe entre le récepteur α2A adrénergique (α2AAR), Gαi1, le dimère Gβγ ainsi que la kinase des récepteurs couplés aux protéines G (GRK2), suite à l’activation du récepteur. De plus, seule l’entrée de GRK2 semble être en mesure de causer la désensibilisation du α2AAR, en s’intercalant entre Gαi1 et Gβγ. Par contre, la stabilisation de l’interaction entre α2AAR et la β-arrestine2 semble nécessiter l’activité kinase de GRK2. Une autre étude a révélé l’importance de différentes Gα pour la mobilisation du calcium, suite à l’activation du récepteur aux opioïdes de type delta (DOR). Suite à la surexpression de Gα de la famille Gαq, il a été possible de mesurer une influence de ces Gα sur la mobilisation du calcium. Toutefois, cette réponse calcique mesurée en présence des Gαq demeure sensible aux prétraitements à la toxine de Bordetella pertussis, qui inhibe sélectivement l’activité des Gαi. De plus, la co-expression de Gαi et Gαq permet de potentialiser la mobilisation de calcium, démontrant une interrelation entre ces deux familles de protéine Gα, pour la signalisation du DOR. Afin de démontrer l’interrelation directe, des expériences de BRET ont été réalisées entre différentes Gα. En plus de montrer la formation de complexes sélectifs entre les Gα, les expériences de BRET réalisées en parallèle d’analyses de séquences de Gα, ont également mis à jour un site de sélectivité d’interaction entre les Gα, l’hélice α4. Suite à la transposition de cette hélice α4 de Gα12 sur Gαi1, qui normalement n’interagissent pas, il a été possible de forcer l’interaction entre Gα12 et Gαi1, confirmant ainsi que cette hélice α contient l’information permettant une sélectivité d’interaction. Au cours de cette thèse, il a été possible de générer de nouvelles méthodes de mesure d’interactions protéiques qui permettent de multiplexer différents signaux, ce qui a permis de mettre à jour de nouvelles interactions entre divers effecteurs de la signalisation de RCGP

Neocartilage formation in 1 g, simulated, and microgravity environments: implications for tissue engineering

The aim of this study was to analyze and compare the deposition of cartilage-specific extracellular matrix components and cellular organization in scaffold-free neocartilage produced in microgravity and simulated microgravity.

Detection of competing DNA structures by thermal gradient gel electrophoresis: from self-association to triple helix formation by (G,A)-containing oligonucleotides

Sequence-specific recognition of DNA can be achieved by triple helix-forming oligonucleotides that bind to the major groove of double-helical DNA. These oligonucleotides have been used as sequence-specific DNA ligands for various purposes, including sequence-specific gene regulation in the so-called ‘antigene strategy’. In particular, (G,A)-containing oligonucleotides can form stable triple helices under physiological conditions. However, triplex formation may be in competition with self-association of these oligonucleotides. For biological applications it would be interesting to identify the conditions under which one structure is favoured as compared to the other(s). Here we have directly studied competition between formation of a parallel (G,A) homoduplex and that of a triple helix by a 13 nt (G,A)-containing oligonucleotide. Temperature gradient gel electrophoresis allows simultaneous detection of competition between the two structures, because of their different temperature dependencies and gel electrophoretic mobilities, and characterisation of this competition.

Étude de la régulation de la protéine G monomérique Rac1 par le facteur de l'ADP-ribosylation 6, lors de la formation d'ondulations de membrane et l'induction de la migration cellulaire

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.