49 research outputs found

    Evaluation of the pocH-100iV DIFF hematology analyzer for use in horses and cattle

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    The results of the analysis of equine and bovine blood samples with the automated pocH-100iV DIFF hematology analyzer were compared with the results obtained with reference methods or other analyzers (Vet ABC, Coulter Counter ZF, Coulter LH 750 analyzer). For equine blood and most parameters in bovine blood good to excellent correlations between methods and analyzers were obtained. For bovine blood good to poor correlations and significant differences were obtained between the pocH-100iV DIFF and other methods or analyzers mainly for hematocrit and hemoglobin determinations and platelet counts. Overall the pocH-100iV DIFF seems to be a reliable and user-friendly analyzer

    Efficient and Specific Internal Cleavage of a Retroviral Palindromic DNA Sequence by Tetrameric HIV-1 Integrase

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    BACKGROUND: HIV-1 integrase (IN) catalyses the retroviral integration process, removing two nucleotides from each long terminal repeat and inserting the processed viral DNA into the target DNA. It is widely assumed that the strand transfer step has no sequence specificity. However, recently, it has been reported by several groups that integration sites display a preference for palindromic sequences, suggesting that a symmetry in the target DNA may stabilise the tetrameric organisation of IN in the synaptic complex. METHODOLOGY/PRINCIPAL FINDINGS: We assessed the ability of several palindrome-containing sequences to organise tetrameric IN and investigated the ability of IN to catalyse DNA cleavage at internal positions. Only one palindromic sequence was successfully cleaved by IN. Interestingly, this symmetrical sequence corresponded to the 2-LTR junction of retroviral DNA circles-a palindrome similar but not identical to the consensus sequence found at integration sites. This reaction depended strictly on the cognate retroviral sequence of IN and required a full-length wild-type IN. Furthermore, the oligomeric state of IN responsible for this cleavage differed from that involved in the 3'-processing reaction. Palindromic cleavage strictly required the tetrameric form, whereas 3'-processing was efficiently catalysed by a dimer. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that the restriction-like cleavage of palindromic sequences may be a general physiological activity of retroviral INs and that IN tetramerisation is strongly favoured by DNA symmetry, either at the target site for the concerted integration or when the DNA contains the 2-LTR junction in the case of the palindromic internal cleavage

    Genetic Burden of TNNI3K in Diagnostic Testing of Patients With Dilated Cardiomyopathy and Supraventricular Arrhythmias

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    BACKGROUND: Genetic variants in TNNI3K (troponin-I interacting kinase) have previously been associated with dilated cardiomyopathy (DCM), cardiac conduction disease, and supraventricular tachycardias. However, the link between TNNI3K variants and these cardiac phenotypes shows a lack of consensus concerning phenotype and protein function. METHODS: We describe a systematic retrospective study of a cohort of patients undergoing genetic testing for cardiac arrhythmias and cardiomyopathy including TNNI3K. We further performed burden testing of TNNI3K in the UK Biobank. For 2 novel TNNI3K variants, we tested cosegregation. TNNI3K kinase function was estimated by TNNI3K autophosphorylation assays.RESULTS: We demonstrate enrichment of rare coding TNNI3K variants in DCM patients in the Amsterdam cohort. In the UK Biobank, we observed an association between TNNI3K missense (but not loss-of-function) variants and DCM and atrial fibrillation. Furthermore, we demonstrate genetic segregation for 2 rare variants, TNNI3K-p.Ile512Thr and TNNI3K-p.His592Tyr, with phenotypes consisting of DCM, cardiac conduction disease, and supraventricular tachycardia, together with increased autophosphorylation. In contrast, TNNI3K-p.Arg556_Asn590del, a likely benign variant, demonstrated depleted autophosphorylation. CONCLUSIONS: Our findings demonstrate an increased burden of rare coding TNNI3K variants in cardiac patients with DCM. Furthermore, we present 2 novel likely pathogenic TNNI3K variants with increased autophosphorylation, suggesting that enhanced autophosphorylation is likely to drive pathogenicity.</p

    Expression of SV2 isoforms during rodent brain development

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    BACKGROUND: SV2A, SV2B and SV2C are synaptic vesicle proteins that are structurally related to members of the major facilitator superfamily (MFS). The function and transported substrate of the SV2 proteins is not clearly defined although they are linked to neurotransmitters release in a presynaptic calcium concentration-dependent manner. SV2A and SV2B exhibit broad expression in the central nervous system while SV2C appears to be more restricted in defined areas such as striatum. SV2A knockout mice start to display generalized seizures at a late developmental stage, around post-natal day 7 (P7), and die around P15. More recently, SV2A was demonstrated to be the molecular target of levetiracetam, an approved anti-epileptic drug (AED). The purpose of this work was to precisely analyze and quantify the SV2A, SV2B and SV2C expression during brain development to understand the contribution of these proteins in brain development and their impact on epileptic seizures. RESULTS: First, we systematically analyzed by immunohistofluorescence, the SV2A, SV2B and SV2C expression during mouse brain development, from embryonic day 12 (E12) to P30. This semi-quantitative approach suggests a modulation of SV2A and SV2B expression in hippocampus around P7. This is the reason why we used various quantitative approaches (laser microdissection of whole hippocampus followed by qRT-PCR and western blot analysis) indicating that SV2A and SV2B expression increased between P5 and P7 and remained stable between P7 and P10. Moreover, the increase of SV2A expression in the hippocampus at P7 was mainly observed in the CA1 region while SV2B expression in this region remains stable. CONCLUSIONS: The observed alterations of SV2A expression in hippocampus are consistent with the appearance of seizures in SV2A−/− animals at early postnatal age and the hypothesis that SV2A absence favors epileptic seizures around P7

    Toolbox Chapelets de terrains

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    Avec d’autres villes europĂ©ennes, la RĂ©gion de Bruxelles-Capitale s’est engagĂ©e Ă  transformer durablement son tissu urbain et a pris diffĂ©rentes initiatives en ce sens, notamment la crĂ©ation ou l’adaptation de ses outils urbanistiques. Une Ă©tude cartographique menĂ©e par le facilitateur Quartiers Durables en collaboration avec l’IGEAT (ULB) a pu Ă©tablir que dans les quartiers existants, l’arsenal urbanistique rĂ©gional prĂȘte une attention particuliĂšre au caractĂšre participatif du dĂ©veloppement de l’espace public et que des outils spĂ©cifiques pour le dĂ©veloppement des grandes friches urbaines existent. Mais pour les chapelets de terrains, aucun outil spĂ©cifique n’avait Ă©tĂ© dĂ©veloppĂ© Ă  l’époque par la RĂ©gion.En 2011, le Service Facilitateur s’est concentrĂ© sur cette typologie des chapelets de terrain. Comme prĂ©cĂ©demment (Toolbox ensembles d’immeubles hauts) la mĂ©thode de travail a Ă©tĂ© la suivante: un premier workshop a Ă©tĂ© organisĂ© autour d’un cas d’étude reprĂ©sentatif de cette typologie: le quartier Moensberg Ă  Uccle. ParallĂšlement des recherches ont Ă©tĂ© pratiquĂ©es en vue de trouver des exemples similaires ou comparables ailleurs en Europe. L’ensemble de ce travail et de ces donnĂ©es a alimentĂ© la rĂ©flexion indispensable Ă  la rĂ©daction de la prĂ©sente publication.Cette recherche a Ă©tĂ© rĂ©alisĂ©e dans le cadre d'une convention Facilitateur Quartiers Durables avec Bruxelles Environnement (2011-2012). TĂ©lĂ©chargeable sur le site de Bruxelles Environnementinfo:eu-repo/semantics/publishe
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