Investigating the Role of cAMP-Signaling in the Regulation of Vascular Endothelial Cell Adaptive Responses to Fluid Shear Stress

The circulating blood exerts a force on the vascular endothelium, termed fluid shear stress (FSS), which directly impacts numerous vascular endothelial cell (VEC) functions. For example, high rates of linear and undisturbed (i.e. laminar) blood flow maintains a protective and quiescent VEC phenotype. Meanwhile, deviations in blood flow, which can occur at vascular branchpoints and large curvatures, create areas of low, and/or oscillatory FSS, and promote a pro-inflammatory, pro-thrombotic and hyperpermeable phenotype. Indeed, it is known that these areas are prone to the development of atherosclerotic lesions. Herein, we show that cyclic nucleotide phosphodiesterase (PDE) 4D (PDE4D) activity is increased by FSS in human arterial endothelial cells (HAECs) and that this activation regulates the activity of cAMP-effector protein, Exchange Protein-activated by cAMP-1 (EPAC1), in these cells. Importantly, we also show that these events directly and critically impact HAEC responses to FSS, especially when FSS levels are low. Both morphological events induced by FSS, as measured by changes in cell alignment and elongation in the direction of FSS, and the expression of critical FSS-regulated genes, including Krüppel-like factor 2 (KLF2), endothelial nitric oxide synthase (eNOS) and thrombomodlin (TM), are mediated by EPAC1/PDE4D signaling. At a mechanistic level, we show that EPAC1/PDE4D acts through the vascular endothelial-cadherin (VECAD)/ platelet-cell adhesion molecule-1 (PECAM1)/vascular endothelial growth factor receptor 2 (VEGFR2) mechanosensor to activate downstream signaling though Akt. Given the critical role of PDE4D in mediating these effects, we also investigated the impact of various patterns of FSS on the expression of individual PDE genes in HAECs. Notably, PDE2A was significantly upregulated in response to high, laminar FSS, while PDE3A was upregulated under low, oscillatory FSS conditions only. These data may provide novel therapeutic targets to limit FSS-dependent endothelial cell dysfunction (ECD) and atherosclerotic development.

Evidence for a Myotomal Hox/Myf Cascade Governing Nonautonomous Control of Rib Specification within Global Vertebral Domains

Hox genes are essential for the patterning of the axial skeleton. Hox group 10 has been shown to specify the lumbar domain by setting a rib-inhibiting program in the presomitic mesoderm (PSM). We have now produced mice with ribs in every vertebra by ectopically expressing Hox group 6 in the PSM, indicating that Hox genes are also able to specify the thoracic domain. We show that the information provided by Hox genes to specify rib-containing and rib-less areas is first interpreted in the myotome through the regional-specific control of Myf5 and Myf6 expression. This information is then transmitted to the sclerotome by a system that includes FGF and PDGF signaling to produce vertebrae with or without ribs at different axial levels. Our findings offer a new perspective of how Hox genes produce global patterns in the axial skeleton and support a redundant nonmyogenic role of Myf5 and Myf6 in rib formation.

Stable isotope record of foraminifera from South Atlantic sediments with reconstruction of paleotemperatures and paleosalinities

Changes in surface water hydrography in the Southern Ocean (eastern Atlantic sector) could be reconstructed on the basis of isotope-geochemical and micropaleontological studies. A total of 75 high quality multicorer sediment surface samples from the southern South Atlantic Ocean and three Quaternary sediment cores, taken on a meridional transect across the Antarctic Circumpolar Current, have been investigated. The results of examining stable oxygen isotope compositions of 24 foraminiferal species and morphotypes were compared to the near-surface hydrography. The different foraminifera have been divided into four groups living at different depths in the upper water column. The 8180 differences between shallow-living (e.g. G. bulloides, N. pachyderma) and deeper-dwelling (e. g. G. inflata) species reflect the measured temperature gradient of the upper 250 m in the water column. Thus, the 6180 difference between shallow-living and deeper-living foraminifera can be used as an indicator for the vertical temperature gradient in the surface water of the Antarctic Circumpolar Current, which is independent of ice volume. All planktonic foraminifera in the surface sediment samples have been counted. 27 species and morphotypes have been selected, to form a reference data Set for statistical purposes. By using R- and Q-mode principal component analysis these planktonic foraminifera have been divided into four and five assemblages, respectively. The geographic distribution of these assemblages is mainly linked to the temperature of sea-surface waters. The five assemblages (factors) of the Q-mode principal component analysis account for 97.l % of the variance of original data. Following the transferfunction- technique a multiple regression between the Q-mode factors and the actual mean sea-surface environmental parameters resulted in a set of equations. The new transfer function can be used to estimate past sea-surface seasonal temperatures for paleoassemblages of planktonic foraminifera with a precision of approximately ±1.2°C. This transfer function F75-27-5 encompasses in particular the environmental conditions in the Atlantic sector of the Antarctic Circumpolar Current. During the last 140,000 years reconstructed sea-surface temperatures fluctuated in the present northern Subantarctic Zone (PS2076-1/3) at an amplitude of up to 7.5°C in summer and of up to 8.5°C in winter. In the present Polarfrontal Zone (PS1754-1) these fluctuations between glacials and interglacials show lower temperatures from 2.5 to 8.5°C in summer and from 1.0 to 5.0°C in winter, respectively. Compared to today, calculated oxygen isotope temperature gradients in the present Subantarctic Zone were lower during the last 140,000 years. This is an indicator for a good mixing of the upper water column. In the Polarfrontal Zone also lower oxygen isotope temperature gradients were found for the glacials 6, 4 and 2. But almost similar temperature gradients as today were found during the interglacial stages 5, 3 and the Holocene, which implicates a mixing of the upper water column compared to present. Paleosalinities were reconstructed by combining d18O-data and the evaluated transfer function paleotemperatures. Especially in the present Polarfrontal Zone (PS1754-1) and in the Antarctic Zone (PS1768-8), a short-term reduction of salinity up to 4 %o, could be detected. This significant reduction in sea-surface water salinity indicates the increased influx of melt-water at the beginning of deglaciation in the southern hemisphere at the end of the last glacial, approximately 16,500-13,000 years ago. The reconstruction of environmental Parameters indicates only small changes in the position of the frontal Systems in the eastern sector of the Antarctic Circumpolar Current during the last 140,000 years. The average position of the Subtropical Front and Subantarctic Front shifted approximately three latitudes between interglacials and glacials. The Antarctic Polar Front shifted approximately four latitudes. But substantial modifications of this scenario have been interpreted for the reconstruction of cold sea-surface temperatures at 41Â S during the oxygen isotope stages 16 and 14 to 12. During these times the Subtropical Front was probably shified up to seven latitudes northwards.

Benthic foraminifera in the Arctic Ocean

We examine the quantitative composition of benthic foraminiferal assemblages of Rose Bengal-stained surface samples from 37 stations in the Laptev Sea, and combine this data set with an existing data set along a transect from Spitsbergen to the central Arctic Ocean. Foraminiferal test accumulation rates, diversity, faunal composition and statistically defined foraminiferal associations are analysed for living (Rose Bengal-stained) and dead foraminifers. We compare the results of several benthic foraminiferal diversity indices and statistically defined foraminiferal associations, including Fisher's alpha and Shannon-Wiener diversity indices, Q-mode principal component analysis and correspondence analysis. Diversity and faunal density (standing stock) of living benthic foraminifers are positively correlated to trophic resources. In contrast, the accumulation rate of dead foraminifers (BFAR) shows fluctuating values depending on test disintegration processes. Foraminiferal associations defined by Q-mode principal component analysis and correspondence analysis are comparable. The factor values of the correspondence analysis allow a quantitative correlation between the foraminiferal fauna and the local carbon flux, which may be used as a tool to estimate changes in primary productivity.

Systematic characterization of the zinc-finger-containing proteins in the mouse transcriptome

Zinc-finger-containing proteins can be classified into evolutionary and functionally divergent protein families that share one or more domains in which a zinc ion is tetrahedrally coordinated by cysteines and histidines. The zinc finger domain defines one of the largest protein superfamilies in mammalian genomes; 46 different conserved zinc finger domains are listed in InterPro (http://www.ebi.ac.uk/InterPro). Zinc finger proteins can bind to DNA, RNA, other proteins, or lipids as a modular domain in combination with other conserved structures. Owing to this combinatorial diversity, different members of zinc finger superfamilies contribute to many distinct cellular processes, including transcriptional regulation, mRNA stability and processing, and protein turnover. Accordingly, mutations of zinc finger genes lead to aberrations in a broad spectrum of biological processes such as development, differentiation, apoptosis, and immunological responses. This study provides the first comprehensive classification of zinc finger proteins in a mammalian transcriptome. Specific detailed analysis of the SP/Kruppel-like factors and the E3 ubiquitin-ligase RING-H2 families illustrates the importance of such an analysis for a more comprehensive functional classification of large protein families. We describe the characterization of a new family of C2H2 zinc-finger-containing proteins and a new conserved domain characteristic of this family, the identification and characterization of Sp8, a new member of the Sp family of transcriptional regulators, and the identification of five new RING-H2 proteins.

Efficacy of sertraline in a 12-week trial for generalized anxiety disorder

Objective: Sertraline's efficacy and tolerability in treating generalized anxiety disorder were evaluated. Method: Adult outpatients with DSM-IV generalized anxiety disorder and a total score of 18 or higher on the Hamilton Anxiety Rating Scale were eligible. After a 1-week single-blind placebo lead-in, patients were randomly assigned to 12 weeks of double-blind treatment with placebo (N=188, mean baseline anxiety score=25) or flexible doses (50-150 mg/day) of sertraline (N=182, mean anxiety score=25). The primary outcome measure was baseline-to-endpoint change in the Hamilton anxiety scale total score. A secondary efficacy measure was the Clinical Global Impression (CGI) improvement score; response was defined as a score of 2 or less. Results: Sertraline patients had significantly greater improvement than placebo patients on all efficacy measures at week 4. Analysis of covariance of the intent-to-treat group at endpoint (with the last observation carried forward) showed a significant difference in the decrease from baseline of the least-square mean total score on the Hamilton anxiety scale between sertraline (mean=11.7) and placebo (mean=8.0). Significantly greater endpoint improvement with sertraline than placebo was obtained for mean scores on the Hamilton anxiety scale psychic factor (6.7 versus 4.1) and somatic factor (5.0 versus 3.9). The rate of responders, based on CGI improvement and last observation carried forward, was significantly higher for sertraline (63%) than placebo (37%). Sertraline was well tolerated; 8% of patients versus 10% for placebo dropped out because of adverse events. Conclusions: Sertraline appears to be efficacious and well tolerated in the treatment of generalized anxiety disorder.

Caractérisation d'un membre de la famille XKLF dans le développement cardiaque

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

Les ligands du récepteur au CNTF dans le système immunitaire

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Caractérisation d'un membre de la famille XKLF dans le développement cardiaque

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

Les ligands du récepteur au CNTF dans le système immunitaire

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.