Proteoglycan Mimic of the Glycocalyx to Treat Endothelial Dysfunction

Patients with kidney failure usually undergo hemodialysis, a process by which toxins produced by the body are filtered from the blood, in order to survive. The preferred form for vascular access is called an arteriovenousfistula (AVF), a surgically created connection between an artery and vein that is utilized to undergo dialysis. However, AVFs have a failure rate of 50-60%. One of the contributions to AVF failure is endothelial cell dysfunction and loss of glycocalyx, which allows neutrophils and other native cells into the media of the vessel, which causes an inflammatory response. Our lab addresses endothelial dysfunction by mimicking the function of the glycocalyx to prevent transmigration of inflammatory cells and ultimately create a healthier vessel for hemodialysis. We have synthesized several glycocalyx mimics consisting of a dermatan sulfate backbone with multiple selectin and ICAM-binding peptides attached. Initial testing involved determining the ability of the variants to bind to inflamed endothelial cells. We also cultured human promyelocytic leukemia cells (HL60) and used retinoic acid to differentiate them into neutrophils. These cells would then test the glycocalyx mimics ability to prevent migration of neutrophils. Thus far, we have seen that the glycocalyx mimics binding to endothelial cells and that this binding is dependent upon the type of selectin and/or ICAM-binding peptides as well as how many peptides are present per dermatan sulfate backbone. We have also shown that proliferation occurs 10 days after seeding, and that rentinoic acid (RA) differentiates HL60 cells into neutrophils. We have developed a protocol for differentiation of HL60 cells to neutrophils, a promising set of glycocalyx mimics, and culturing method for HL60 cells

Overview of bedrock mapping in the northern and western parts of the Tehery Lake-Wager Bay area, western Hudson Bay, Nunavut

Bedrock-geology mapping was conducted in the summer of 2016 in the Tehery Lake–Wager Bay area on the northwestern coast of Hudson Bay, Nunavut, as part of a multiyear, multidisciplinary mapping campaign led by the Geological Survey of Canada, through Phase 2 of the Geo-mapping for Energy a nd Minerals program (GEM-2), and the Canada-Nunavut Geoscience Office. Fieldwork resulted in the identification and spatial constraint of rock units in the northern and western parts of the study area, which were sampled for geochemical, g eochronological and petrographic analysis, as well as to as- sess their economic potential. Mapping has revealed the presence of a large granulite-facies metamorphic domain in the southern part of the study area; the possibility of two different supracrustal rock sequences; the western continuations of the Chesterfield fault zone and Wager shear zone; and generally high, but locally variable, peak metamorphic conditions across the study area. Further analytical work is required to fully characterize rock units, compare and correlate them with other well-studied units, and determine the geological history and economic potential of the Tehery Lake–Wager Bay area

Toward Automation of the Supine Pressor Test for Preeclampsia

Preeclampsia leads to increased risk of morbidity and mortality for both mother and fetus. Most previous studies have largely neglected mechanical compression of the left renal vein by the gravid uterus as a potential mechanism. In this study, we first used a murine model to investigate the pathophysiology of left renal vein constriction. The results indicate that prolonged renal vein stenosis after 14 days can cause renal necrosis and an increase in blood pressure (BP) of roughly 30 mmHg. The second part of this study aimed to automate a diagnostic tool, known as the supine pressor test (SPT), to enable pregnant women to assess their preeclampsia development risk. A positive SPT has been previously defined as an increase of at least 20 mmHg in diastolic BP when switching between left lateral recumbent and supine positions. The results from this study established a baseline BP increase between the two body positions in nonpregnant women and demonstrated the feasibility of an autonomous SPT in pregnant women. Our results demonstrate that there is a baseline increase in BP of roughly 10-14 mmHg and that pregnant women can autonomously perform the SPT. Overall, this work in both rodents and humans suggests that (1) stenosis of the left renal vein in mice leads to elevation in BP and acute renal failure, (2) nonpregnant women experience a baseline increase in BP when they shift from left lateral recumbent to supine position, and (3) the SPT can be automated and used autonomously

Lithotectonic Framework of the Core Zone, Southeastern Churchill Province, Canada

The Core Zone, a broad region located between the Superior and North Atlantic cratons and predominantly underlain by Archean gneiss and granitoid rocks, remained until recently one of the less well known parts of the Canadian Shield. Previously thought to form part of the Archean Rae Craton, and later referred to as the Southeastern Churchill Province, it has been regarded as an ancient continental block trapped between the Paleoproterozoic Torngat and New Quebec orogens, with its relationships to the adjacent Superior and North Atlantic cratons remaining unresolved. The geochronological data presented herein suggest that the Archean evolution of the Core Zone was distinct from that in both the Superior and North Atlantic (Nain) cratons. Moreover, the Core Zone itself consists of at least three distinct lithotectonic entities with different evolutions, referred to herein as the George River, Mistinibi-Raude and Falcoz River blocks, that are separated by steeply-dipping, crustal-scale shear zones interpreted as paleosutures. Specifically, the George River Block consists of ca. 2.70 Ga supracrustal rocks and associated ca. 2.70–2.57 Ga intrusions. The Mistinibi-Raude Block consists of remnants of a ca. 2.37 Ga volcanic arc intruded by a ca. 2.32 Ga arc plutonic suite (Pallatin) and penecontemporaneous alkali plutons (Pelland and Nekuashu suites). It also hosts a coarse clastic cover sequence (the Hutte Sauvage Group) which contains detrital zircons provided from locally-derived, ca. 2.57–2.50 Ga, 2.37–2.32 Ga, and 2.10–2.08 Ga sources, with the youngest concordant grain dated at 1987 ± 7 Ma. The Falcoz River Block consists of ca. 2.89–2.80 Ga orthogneiss intruded by ca. 2.74–2.70 granite, tonalite, and granodiorite. At the western margin of the Core Zone, the George River Block and Kuujjuaq Domain may have been proximal by ca. 1.84 Ga as both appear to have been sutured by the 1.84–1.82 Ga De Pas Batholith, whereas at its eastern margin, the determination of metamorphic ages of ca. 1.85 to 1.80 Ga in the Falcoz River Block suggests protracted interaction with the adjacent Lac Lomier Complex during their amalgamation and suturing, but with a younger, ‘New Quebec’ overprint as well. The three crustal blocks forming the Core Zone add to a growing list of ‘exotic’ Archean to earliest Paleoproterozoic microcontinents and crustal slices that extend around the Superior Craton from the Grenville Front through Hudson Strait, across Hudson Bay and into Manitoba and Saskatchewan, in what was the Manikewan Ocean realm, which closed between ca. 1.83–1.80 Ga during the formation of supercontinent Nuna.RÉSUMÉLa Zone noyau, une vaste région située entre les cratons du Supérieur et de l’Atlantique Nord et reposant principalement sur des gneiss archéens et des roches granitiques, est demeurée jusqu’à récemment l’une des parties les moins bien connues du Bouclier canadien. Considérée auparavant comme faisant partie du craton archéen de Rae, puis comme la portion sud-est de la Province de Churchill, on l’a perçue comme un ancien bloc continental piégé entre les orogènes paléoprotérozoïques des Torngat et du Nouveau-Québec, ses relations avec les cratons supérieurs adjacents et de l’Atlantique Nord demeurant nébuleuses. Les données géochronologiques présentées ici permettent de penser que l’évolution archéenne de la Zone noyau a été différente de celle des cratons du Supérieur et de l’Atlantique Nord (Nain). De plus, la Zone noyau elle-même se compose d’au moins trois entités lithotectoniques distinctes avec des évolutions différentes, appelées ici les blocs de la rivière George, de Mistinibi-Raude et de la rivière Falcoz, lesquels sont séparées par des zones de cisaillement crustales à forte inclinaison, conçues comme des paléosutures. Plus précisément, le bloc de la rivière George est constitué de roches supracrustales d'env. 2,70 Ga, et d’intrusions connexes d'env. 2,70–2,57 Ga. Le bloc Mistinibi-Raude est constitué de vestiges d’un arc volcanique d'env. 2,37 Ga, recoupé par une suite plutonique d’arc d'env. 2,32 Ga (Pallatin) et de plutons alcalins péné-contemporains (suites Pelland et Nekuashu). Il contient également une séquence de couverture clastique grossière (le groupe Hutte Sauvage) renfermant des zircons détritiques de sources locales, âgés d'env. 2,57–2,50 Ga, 2,37–2,32 Ga et 2,10–2,08 Ga, le grain concordant le plus jeune étant âgé de 1987 ± 7 Ma. Le bloc de la rivière Falcoz est formé d’un orthogneiss âgé d'env. 2,89–2,80 Ga, recoupé par des intrusions de granite, tonalite et granodiorite âgées d'env. 2,74–2,70 Ga. À la marge ouest de la Zone noyau, le bloc de la rivière George et du domaine de Kuujjuaq peuvent avoir été proximaux il y a 1,84 Ga env., car les deux semblent avoir été suturés par le batholithe De Pas il y a environ 1,84–1,82 Ga, alors qu’à sa marge est, la détermination des datations métamorphiques de 1,85 à 1,80 Ga dans le bloc de la rivière Falcoz suggère une interaction prolongée avec le complexe adjacent du lac Lomier durant leur amalgamation et leur suture, mais affecté aussi d’une surimpression « Nouveau Québec » plus jeune. Les trois blocs crustaux formant la Zone noyau s’ajoutent à une liste croissante de micro-continents et d’écailles crustales « exotiques » archéennes à paléoprotérozoïques très précoces qui s’étalent autour du craton Supérieur depuis le front de Grenville jusqu’au Manitoba, à travers le détroit d’Hudson, la baie d’Hudson jusque dans le Manitoba et la Saskatchewan, là où s’étendait l’océan Manikewan, lequel s’est refermé il y a environ 1,83–1,80 Ga, pendant la formation du supercontinent Nuna

Physico-chemical foundations underpinning microarray and next-generation sequencing experiments

Hybridization of nucleic acids on solid surfaces is a key process involved in high-throughput technologies such as microarrays and, in some cases, next-generation sequencing (NGS). A physical understanding of the hybridization process helps to determine the accuracy of these technologies. The goal of a widespread research program is to develop reliable transformations between the raw signals reported by the technologies and individual molecular concentrations from an ensemble of nucleic acids. This research has inputs from many areas, from bioinformatics and biostatistics, to theoretical and experimental biochemistry and biophysics, to computer simulations. A group of leading researchers met in Ploen Germany in 2011 to discuss present knowledge and limitations of our physico-chemical understanding of high-throughput nucleic acid technologies. This meeting inspired us to write this summary, which provides an overview of the state-of-the-art approaches based on physico-chemical foundation to modeling of the nucleic acids hybridization process on solid surfaces. In addition, practical application of current knowledge is emphasized

Whole genome analysis of a wine yeast strain

Saccharomyces cerevisiae strains frequently exhibit rather specific phenotypic features needed for adaptation to a special environment. Wine yeast strains are able to ferment musts, for example, while other industrial or laboratory strains fail to do so. The genetic differences that characterize wine yeast strains are poorly understood, however. As a first search of genetic differences between wine and laboratory strains, we performed DNA-array analyses on the typical wine yeast strain T73 and the standard laboratory background in S288c. Our analysis shows that even under normal conditions, logarithmic growth in YPD medium, the two strains have expression patterns that differ significantly in more than 40 genes. Subsequent studies indicated that these differences correlate with small changes in promoter regions or variations in gene copy number. Blotting copy numbers vs. transcript levels produced patterns, which were specific for the individual strains and could be used for a characterization of unknown samples. Copyright © 2001 John Wiley & Sons, Ltd

Vibration Response Imaging: evaluation of rater agreement in healthy subjects and subjects with pneumonia

<p>Abstract</p> <p>Background</p> <p>We evaluated pulmonologists variability in the interpretation of Vibration response imaging (VRI) obtained from healthy subjects and patients hospitalized for community acquired pneumonia.</p> <p>Methods</p> <p>The present is a prospective study conducted in a tertiary university hospital. Twenty healthy subjects and twenty three pneumonia cases were included in this study. Six pulmonologists blindly analyzed images of normal subjects and pneumonia cases and evaluated different aspects of VRI images related to the quality of data aquisition, synchronization of the progression of breath sound distribution and agreement between the maximal energy frame (MEF) of VRI (which is the maximal geographical area of lung vibrations produced at maximal inspiration) and chest radiography. For qualitative assessment of VRI images, the raters' evaluations were analyzed by degree of consistency and agreement.</p> <p>Results</p> <p>The average value for overall identical evaluations of twelve features of the VRI image evaluation, ranged from 87% to 95% per rater (94% to 97% in control cases and from 79% to 93% per rater in pneumonia cases). Inter-rater median (IQR) agreement was 91% (82-96). The level of agreement according to VRI feature evaluated was in most cases over 80%; intra-class correlation (ICC) obtained by using a model of subject/rater for the averaged features was overall 0.86 (0.92 in normal and 0.73 in pneumonia cases).</p> <p>Conclusions</p> <p>Our findings suggest good agreement in the interpretation of VRI data between different raters. In this respect, VRI might be helpful as a radiation free diagnostic tool for the management of pneumonia.</p

Molecular and enological characterization of a natural Saccharomyces uvarum and Saccharomyces cerevisiae hybrid

Saccharomyces cerevisiae plays a main role in the winemaking process, although other species, like Saccharomyces uvarum or Saccharomyces paradoxus, have been associated with must fermentations. It has been reported in recent years, that yeast hybrids of different Saccharomyces species might be responsible for wine productions. Although S. cerevisiae × Saccharomyces kudriavzevii hybrids have been well studied, very little attention has been paid to S. cerevisiae × S. uvarum hybrids. In this work we characterized the genomic composition of S6U, a widely used commercial S. cerevisiae × S. uvarum yeast hybrid isolated in wine fermentations containing one copy of the genome of each parental species, which suggests a relatively recent hybridization event. We also studied its performance under diverse enological conditions. The results show enhanced performance under low temperature enological conditions, increased glycerol production, lower acetic acid production and increased production of interesting aroma compounds. We also examined the transcriptomic response of the S6U hybrid strain compared with the reference species under enological conditions. The results show that although the hybrid strain transcriptome is more similar to S. uvarum than to S. cerevisiae, it presents specifically regulated genes involved in stress response, lipids and amino acid metabolism. The enological performance and aroma profile of this S. cerevisiae × S. uvarum hybrid makes it a good candidate for participating in winemaking, especially at low temperatures

Exploiting Chemical Libraries, Structure, and Genomics in the Search for Kinase Inhibitors

Selective protein kinase inhibitors were developed on the basis of the unexpected binding mode of 2,6,9-trisubstituted purines to the adenosine triphosphate-binding site of the human cyclin-dependent kinase 2 (CDK2). By iterating chemical library synthesis and biological screening, potent inhibitors of the human CDK2-cyclin A kinase complex and of Saccharomyces cerevisiae Cdc28p were identified. The structural basis for the binding affinity and selectivity was determined by analysis of a three-dimensional crystal structure of a CDK2-inhibitor complex. The cellular effects of these compounds were characterized in mammalian cells and yeast. In the latter case the effects were characterized on a genome-wide scale by monitoring changes in messenger RNA levels in treated cells with high-density oligonucleotide probe arrays. Purine libraries could provide useful tools for analyzing a variety of signaling and regulatory pathways and may lead to the development of new therapeutics

VEGF165-induced vascular permeability requires NRP1 for ABL-mediated SRC family kinase activation.

The vascular endothelial growth factor (VEGF) isoform VEGF165 stimulates vascular growth and hyperpermeability. Whereas blood vessel growth is essential to sustain organ health, chronic hyperpermeability causes damaging tissue edema. By combining in vivo and tissue culture models, we show here that VEGF165-induced vascular leakage requires both VEGFR2 and NRP1, including the VEGF164-binding site of NRP1 and the NRP1 cytoplasmic domain (NCD), but not the known NCD interactor GIPC1. In the VEGF165-bound receptor complex, the NCD promotes ABL kinase activation, which in turn is required to activate VEGFR2-recruited SRC family kinases (SFKs). These results elucidate the receptor complex and signaling hierarchy of downstream kinases that transduce the permeability response to VEGF165. In a mouse model with choroidal neovascularisation akin to age-related macular degeneration, NCD loss attenuated vessel leakage without affecting neovascularisation. These findings raise the possibility that targeting NRP1 or its NCD interactors may be a useful therapeutic strategy in neovascular disease to reduce VEGF165-induced edema without compromising vessel growth