Synthetic mRNA Encoding VEGF-A in Patients Undergoing Coronary Artery Bypass Grafting: Design of a Phase 2a Clinical Trial

Therapeutic angiogenesis may improve outcomes in patients with coronary artery disease undergoing surgical revascularization. Angiogenic factors may promote blood vessel growth and regenerate regions of ischemic but viable myocardium. Previous clinical trials of vascular endothelial growth factor A (VEGF-A) gene therapy with DNA or viral vectors demonstrated safety but not efficacy. AZD8601 is VEGF-A165 mRNA formulated in biocompatible citrate-buffered saline and optimized for high-efficiency VEGF-A expression with minimal innate immune response. EPICCURE is an ongoing randomized, double-blind, placebo-controlled study of the safety of AZD8601 in patients with moderately decreased left ventricular function (ejection fraction 30%–50%) undergoing elective coronary artery bypass surgery. AZD8601 3 mg, 30 mg, or placebo is administered as 30 epicardial injections in a 10-min extension of cardioplegia. Injections are targeted to ischemic but viable myocardial regions in each patient using quantitative 15O-water positron emission tomography (PET) imaging (stress myocardial blood flow 0.6 mL/g/min). Improvement in regional and global myocardial blood flow quantified with 15O-water PET is an exploratory efficacy outcome, together with echocardiographic, clinical, functional, and biomarker measures. EPICCURE combines high-efficiency delivery with quantitative targeting and follow-up for robust assessment of the safety and exploratory efficacy of VEGF-A mRNA angiogenesis (ClinicalTrials.gov: NCT03370887).</p

Design and rationale of FLAVOUR: A phase IIa efficacy study of the 5-lipoxygenase activating protein antagonist AZD5718 in patients with recent myocardial infarction

levels is the primary efficacy outcome. FLAVOUR also aims to evaluate whether AZD5718 can improve coronary microvascular function, as measured by transthoracic colour Doppler-assisted coronary flow velocity reserve. Centrally pretrained study sonographers use standardized protocols and equipment. Additional outcomes include assessment of comprehensive echocardiographic parameters (including coronary flow, global strain, early diastolic strain rate and left ventricular ejection fraction), arterial stiffness, biomarkers, health-related quality of life, and safety. Specific anti-inflammatory therapies may represent novel promising treatments to reduce residual risk in patients with coronary artery disease. By combining primary pharmacodynamic and secondary cardiovascular surrogate efficacy outcomes, FLAVOUR aims to investigate the mechanistic basis and potential benefits of AZD5718 treatment in patients with coronary artery disease

DNA Display Selection of Peptide Ligands for a Full-Length Human G Protein-Coupled Receptor on CHO-K1 Cells

The G protein-coupled receptors (GPCRs), which form the largest group of transmembrane proteins involved in signal transduction, are major targets of currently available drugs. Thus, the search for cognate and surrogate peptide ligands for GPCRs is of both basic and therapeutic interest. Here we describe the application of an in vitro DNA display technology to screening libraries of peptide ligands for full-length GPCRs expressed on whole cells. We used human angiotensin II (Ang II) type-1 receptor (hAT1R) as a model GPCR. Under improved selection conditions using hAT1R-expressing Chinese hamster ovary (CHO)-K1 cells as bait, we confirmed that Ang II gene could be enriched more than 10,000-fold after four rounds of selection. Further, we successfully selected diverse Ang II-like peptides from randomized peptide libraries. The results provide more precise information on the sequence-function relationships of hAT1R ligands than can be obtained by conventional alanine-scanning mutagenesis. Completely in vitro DNA display can overcome the limitations of current display technologies and is expected to prove widely useful for screening diverse libraries of mutant peptide and protein ligands for receptors that can be expressed functionally on the surface of CHO-K1 cells

The Adhesion GPCR GPR125 is specifically expressed in the choroid plexus and is upregulated following brain injury

<p>Abstract</p> <p>Background</p> <p>GPR125 belongs to the family of <it>Adhesion </it>G protein-coupled receptors (GPCRs). A single copy of GPR125 was found in many vertebrate genomes. We also identified a <it>Drosophila </it>sequence, DmCG15744, which shares a common ancestor with the entire Group III of <it>Adhesio</it>n GPCRs, and also contains Ig, LRR and HBD domains which were observed in mammalian GPR125.</p> <p>Results</p> <p>We found specific expression of GPR125 in cells of the choroid plexus using <it>in situ </it>hybridization and protein-specific antibodies and combined <it>in situ</it>/immunohistochemistry co-localization using cytokeratin, a marker specific for epithelial cells. Induction of inflammation by LPS did not change GPR125 expression. However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.</p> <p>Conclusion</p> <p>These findings suggest that GPR125 plays a functional role in choroidal and hippocampal response to injury.</p

4-aminobutyrate aminotrasferase (ABAT): genetic and pharmacological evidence for an involvement in gastro esophageal reflux disease

Extent: 9p.Gastro-esophageal reflux disease (GERD) is partly caused by genetic factors. The underlying susceptibility genes are currently unknown, with the exception of COL3A1. We used three independent GERD patient cohorts to identify GERD susceptibility genes. Thirty-six families, demonstrating dominant transmission of GERD were subjected to whole genome microsatellite genotyping and linkage analysis. Five linked regions were identified. Two families shared a linked region (LOD 3.9 and 2.0) on chromosome 16. We used two additional independent GERD patient cohorts, one consisting of 219 trios (affected child with parents) and the other an adult GERD case control cohort consisting of 256 cases and 485 controls, to validate individual genes in the linked region through association analysis. Sixty six single nucleotide polymorphism (SNP) markers distributed over the nine genes present in the linked region were genotyped in the independent GERD trio cohort. Transmission disequilibrium test analysis followed by multiple testing adjustments revealed a significant genetic association for one SNP located in an intron of the gene 4-aminobutyrate aminotransferase (ABAT) (Padj = 0.027). This association did not replicate in the adult case-control cohort, possibly due to the differences in ethnicity between the cohorts. Finally, using the selective ABAT inhibitor vigabatrin (c-vinyl GABA) in a dog study, we were able to show a reduction of transient lower esophageal sphincter relaxations (TLESRs) by 57.3611.4 % (p = 0.007) and the reflux events from 3.160.4 to 0.860.4 (p = 0.007). Our results demonstrate the direct involvement of ABAT in pathways affecting lower esophageal sphincter (LES) control and identifies ABAT as a genetic risk factor for GERD.Johan Jirholt, Bengt Åsling, Paul Hammond, Geoffrey Davidson, Mikael Knutsson, Anna Walentinsson, Jörgen M. Jensen, Anders Lehmann, Lars Agreus and Maria Lagerström-Ferme

Occurrence and environmental risk assessment of antifouling paint biocides from leisure boats

The use of biocidal antifouling (AF) paints is the most common way to prevent fouling on leisure boat hulls. The main aim of this thesis was to investigate the pathways through which AF biocides, past and present, may reach the environment through their use on leisure boats and to improve the risk assessment of biocidal AF paints intended for amateur use. The work presented focuses mainly on the Baltic Sea, with emphasis on regulation and risk assessment procedures in Sweden. A new method was developed for the quantification of nowadays banned organotin compounds (OTCs) such as tributyltin (TBT) in paint flakes (paper I). OTCs were detected in hull paint scrapings from three countries around the Baltic Sea. Thus, historic layers of organotin paint on leisure boats may constitute as sources of TBT to the marine environment. Total tin was identified as an indicator for the presence of OTCs on boat hulls, allowing for quicker identification of vessels in need of remediation. Nowadays, most AF paints tend to contain high amounts of copper (Cu) and zinc (Zn). The use of AF paints was shown to cause exceedance of guideline values for these two metals in soil, sediment and water in various investigated marinas (papers II and IV). The pollution of boatyard soil was linked to hull maintenance activities carried out over unprotected ground (paper II). AF paints were also found to impact both the concentration and speciation of dissolved Cu and Zn in two Baltic Sea marinas, with increased concentrations as well as an increased proportion of bioavailable species as a function of an increased number of moored boats (paper IV). A new method utilizing X-Ray Fluorescence (XRF) was used to derive the release rates of Cu and Zn in the field for five commercially available AF paints for amateur use (paper III). Salinity and paint properties were found to be important parameters affecting the release. The in situ release rates were also found to exceed those derived with current standardized release rate methods. Given the high release rates, none of the studied paints should have been approved for the Swedish market. This finding likely explains the exceedance of guideline values for dissolved Cu and Zn in investigated Baltic Sea marinas (paper IV). In conclusion, there is a need for caution when authorizing new biocides as the phasing out of banned substances can be a lengthy process due to their continued presence in historic paint layers. Additionally, paint-specific release rates determined under conditions reflecting the intended use of the product should be used for a more realistic environmental risk assessment of AF paints.At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 4: Manuscript.</p

Anhörigas upplevelse av att vara närvarande vid pågående hjärt- och lungräddning

Abstrakt Bakgrund: I Sverige inträffar ca 10 000 hjärtstopp utanför sjukhus varje år. Ett hjärtstopp är ofta oväntat och många gånger finns det anhöriga i närheten. För att kunna rädda liv krävs en snabb insats med hjärt-och lungräddning. Sjukvårdspersonal arbetar under stress med stort fokus på uppgiften och ser därför inte att det finns en anhörig i närheten. Syftet var att belysa hur anhöriga upplevde att vara närvarande vid pågående hjärt-och lungräddning av nära anhörig inom akutsjukvården. Metod: Studien genomfördes som en systematisk litteraturstudie på 11 vetenskapliga artiklar framsökta i databaser och referenslistor. Resultatet grundades på tre kategorier, delaktighet-att få vara närvarande, information och tidigare kunskap samt psykisk påverkan. Resultatet visade att anhöriga som varit med vid hjärt- och lungräddning klarade av att hantera sin sorg bättre och hade mindre psykosocialpåverkan jämfört med de som inte varit närvarande. De anhöriga önskade information och kände trygghet i att ha en stödperson vid sin sida. Slutsats: Sjukvårdpersonalen måste få ökad trygghet i att våga se och bjuda in de anhöriga vid en akut händelse. Sjukvården kan med relativt enkla medel göra en vinst i att anhöriga känner sig bekräftade och välinformerade och klarar av att hantera sin sorg

Utveckling av analysverktyg för att bedöma påverkan från tributyltenn (TBT) i svenska vattenförekomster

Den tennorganiska föreningen tributyltenn (TBT) tillhör ett av de giftigaste ämnena som människan någonsin släppt ut till vattenmiljön och förbjöds för användning i fritidsbåtsfärger 1989 och sedan 2008 råder ett globalt förbud för fartyg enligt AFS-konventionen. Trots detta fortsätter TBT att spridas till vattenmiljön där hantering av båtskrov, förorenade sediment och fartyg till sjöss bedöms utgöra källor för kontinuerlig tillförsel av TBT till miljön. Syftet med projektet var att försöka ta fram en modell för att prediktera halt av TBT i sediment med hjälp av olika förklaringsparametrar. Det slutgiltiga målet var att med hjälp av modellen därefter kunna identifiera geografiska områden där TBT-halter riskerar att överskrida miljökvalitetsnormen för sediment. Analysdata insamlades från Sveriges Länsstyrelser, SGU, Västerås stad, Stockholm stad samt Nacka kommun. Jämförelse av TBT-halterna i ytsediment (0-2 cm) med gränsvärdet på 1,6 μg/kg TS visar 83 % av alla provpunkter överskrider gränsvärdet. För de marina punkterna skulle eventuellt en fritidsbåtsaktivitet ≥ 45000 båtar kunna användas för att identifiera de vattenförekomster med störst risk för överskridning. 48% av vattenförkomster längs kusten visade sig ha denna egenskap. Analysen har dock stora osäkerheter och en lägre fritidsbåtsaktivitet utesluter inte att miljökvalitetsnormen för TBT överskrids. För att validera hur väl parametern "fritidsbåtsaktivitet &gt; 45000 båtar" predikterar om TBT-halter överskrider miljökvalitetsnormen bör framtida miljöövervakning även inkludera vattenförekomster med en fritidsbåtsaktivitet under 45000 båtar

Creating and sharing knowledge within a transnational team--the development of a global business system

By mean of transnational teams, local knowledge of business units can be leveraged to a global level for the development of common global business solutions in the multinational corporation. We investigate this increasingly valuable means of supporting the creation and sharing of knowledge. Empirical evidence from a case study of a transnational team identifies socialization of team members as perhaps primary for efficient creation and sharing of knowledge, and information technology as secondary.