Granulin Knock Out Zebrafish Lack Frontotemporal Lobar Degeneration and Neuronal Ceroid Lipofuscinosis Pathology

Loss of function mutations in granulin (GRN) are linked to two distinct neurological disorders, frontotemporal lobar degeneration (FTLD) and neuronal ceroid lipofuscinosis (NCL). It is so far unknown how a complete loss of GRN in NCL and partial loss of GRN in FTLD can result in such distinct diseases. In zebrafish, there are two GRN homologues, Granulin A (Grna) and Granulin B (Grnb). We have generated stable Grna and Grnb loss of function zebrafish mutants by zinc finger nuclease mediated genome editing. Surprisingly, the grna and grnb single and double mutants display neither spinal motor neuron axonopathies nor a reduced number of myogenic progenitor cells as previously reported for Grna and Grnb knock down embryos. Additionally, grna-/-;grnb-/- double mutants have no obvious FTLD- and NCL-related biochemical and neuropathological phenotypes. Taken together, the Grna and Grnb single and double knock out zebrafish lack any obvious morphological, pathological and biochemical phenotypes. Loss of zebrafish Grna and Grnb might therefore either be fully compensated or only become symptomatic upon additional challenge

Rat Monoclonal Antibodies Specific for LST1 Proteins

The LST1 gene is located in the human MHC class III region and encodes transmembrane and soluble isoforms that have been suggested to play a role in the regulation of the immune response and are associated with inflammatory diseases such as rheumatoid arthritis. Here we describe the generation and characterization of the first monoclonal antibodies against LST1. Two hybridoma lines secreting monoclonal antibodies designated 7E2 and 8D12 were established. The 7E2 antibody detects recombinant and endogenous LST1 by Western blot analysis while 8D12 reacts with recombinant and endogenous LST1 in immunoprecipitation and flow cytometry procedures. The newly established antibodies were used to survey LST1 protein expression in human cell lines, which was found to be tightly regulated, allowing the expression of transmembrane isoforms but suppressing soluble isoforms

100 Years LANZ Bulldog

Im Jahr 2021 ist es 100 Jahre her, dass der legendäre LANZ Bulldog in seiner Urform der Öffentlichkeit vorgestellt wurde. Das war 1921 auf der DLG-Ausstellung in Leipzig, einem Vorläufer der heutigen AGRITECHNICA. Die erste Ausführung war ein LANZ HL mit 12 PS, der die Mechanisierung der Landwirtschaft signifikant und nachhaltig geprägt hat. Anlässlich seines hundertjährigen Jubiläums will dieser Beitrag die Entwicklung, einige technische Besonderheiten und Anekdoten rund um den LANZ Bulldog herausarbeiten und vorstellen.In 2021 it will be 100 years since the legendary LANZ Bulldog in its initial form was presented to the general public. That was in 1921 at the DLG exhibition in Leipzig, a predecessor of today's AGRITECHNICA fair. The first model was a LANZ HL with 12 hp, which significantly and sustainably influenced the mechanization of agriculture. On the occasion of its centenary, this article aims to highlight and present the development, some technical features and anecdotes related to the LANZ Bulldog

The Protein Kinase KIS Impacts Gene Expression during Development and Fear Conditioning in Adult Mice

The brain-enriched protein kinase KIS (product of the gene UHMK1) has been shown to phosphorylate the human splicing factor SF1 in vitro. This phosphorylation in turn favors the formation of a $U2AF^{65}$-SF1-RNA complex which occurs at the 3′ end of introns at an early stage of spliceosome assembly. Here, we analyzed the effects of KIS knockout on mouse SF1 phosphorylation, physiology, adult behavior, and gene expression in the neonate brain. We found SF1 isoforms are differently expressed in KIS-ko mouse brains and fibroblasts. Re-expression of KIS in fibroblasts restores a wild type distribution of SF1 isoforms, confirming the link between KIS and SF1. Microarray analysis of transcripts in the neonate brain revealed a subtle down-regulation of brain specific genes including cys-loop ligand-gated ion channels and metabolic enzymes. Q-PCR analyses confirmed these defects and point to an increase of pre-mRNA over mRNA ratios, likely due to changes in splicing efficiency. While performing similarly in prepulse inhibition and most other behavioral tests, KIS-ko mice differ in spontaneous activity and contextual fear conditioning. This difference suggests that disregulation of gene expression due to KIS inactivation affects specific brain functions

Portugal, Política Externa e o Luxemburgo: Estudo das Relações Consulares e da Comunidade Portuguesa

No contexto da recente reformulação da política externa portuguesa com a adição de novos vetores estratégicos, dos quais as comunidades portuguesas no estrangeiro, a atividade diplomática adquiriu uma importância redobrada, a nível dos serviços consulares. Nesta investigação, pretende-se analisar o caso da comunidade portuguesa no Grão-Ducado do Luxemburgo. Tendo por base uma contextualização histórica da evolução da política externa portuguesa e da emigração portuguesa, tanto num plano geral como especificamente o caso do Grão-Ducado do Luxemburgo pretende-se analisar a comunidade portuguesa que se estabeleceu neste país. Mais ainda, e compreender de que forma é que a política externa portuguesa, através da diplomacia e, mais especificamente, o seu serviço consular correspondente, o Consulado-Geral de Portugal no Luxemburgo, atua para garantir os direitos e satisfazer as necessidades da comunidade portuguesa no país. Esta análise fundamentalmente teórica é complementada com um relatório de estágio, que decorreu entre 4 de novembro de 2019 e 4 de julho de 2020 no Consulado-Geral de Portugal no Luxemburgo.In the context of the recent reformulation of Portuguese foreign policy with the addition of new strategic vectors, one of them being the Portuguese communities abroad, diplomatic activity has acquired an increased importance in terms of consular services. In this investigation, the case of the Portuguese community in the Grand Duchy of Luxembourg will be the subject of analysis. Based on a historical contextualization of the evolution of Portuguese foreign policy and Portuguese emigration, both generally and specifically in the case of the Grand Duchy of Luxembourg, it is intended to study the Portuguese community that has established itself in this country. Moreover, understanding how Portuguese foreign policy, through diplomacy and, specifically its corresponding consular service, the Consulate General of Portugal in Luxembourg, operates to guarantee the rights and satisfy the needs of the Portuguese community in this country. This fundamentally theoretical analysis will be complemented with an internship report, which took place between November 4th, 2019 and July 4th, 2020, at the Consulate General of Portugal in Luxembourg

Structural instabilities during cyclic loading of ultrafine-grained copper studied with micro bending experiments

The cyclic mechanical properties and microstructural stability of severe plastically deformed copper were investigated by means of micro bending experiments. The ultrafine-grained structure of OFHC copper was synthesized utilizing the high pressure torsion (HPT) technique. Micron sized cantilevers were focused-ion-beam milled and subsequently tested within a scanning electron microscope in the low cycle fatigue regime at strain amplitudes in the range of 1.1 − 3.2 ∗ 10−3. It was found that HPT processed ultra-fine grained copper is prone to cyclic softening, which is a consequence of grain coarsening in the absence of shear banding in the micro samples. Novel insights into the grain coarsening mechanism were revealed by quasi in-situ EBSD scans, showing i) continuous migration of high angle grain boundaries, ii) preferential growth of larger grains at the expense of adjacent smaller ones, iii) a reduction of misorientation gradients within larger grains if the grain structure in the neighborhood is altered and iv) no evidence that a favorable crystallographic orientation drives grain growth during homogeneous coarsening at moderate accumulated strains, tested here

Drosophila HP1c Is Regulated by an Auto-Regulatory Feedback Loop through Its Binding Partner Woc

HP1 is a major component of chromatin and regulates gene expression through its binding to methylated histone H3. Most eukaryotes express at least three isoforms of HP1 with similar domain architecture. However, despite the common specificity for methylated histone H3, the three HP1 isoforms bind to different regions of the genome. Most of the studies so far focused on the HP1a isoform and its role in transcriptional regulation. As HP1a requires additional factors to bind methylated chromatin in vitro, we wondered whether another isoform might also require additional targeting factors. Indeed, we found that HP1c interacts with the DNA binding factors Woc and Row and requires Woc to become targeted to chromatin in vivo. Moreover, we show that the interaction between HP1c and Woc constitutes a transcriptional feedback loop that operates to balance the concentration of HP1c within the cell. This regulation may prevent HP1c from binding to methylated heterochromatin

The putative Notch ligand HyJagged is a transmembrane protein present in all cell types of adult Hydra and upregulated at the boundary between bud and parent

<p>Abstract</p> <p>Background</p> <p>The Notch signalling pathway is conserved in pre-bilaterian animals. In the Cnidarian <it>Hydra </it>it is involved in interstitial stem cell differentiation and in boundary formation during budding. Experimental evidence suggests that in <it>Hydra </it>Notch is activated by presenilin through proteolytic cleavage at the S3 site as in all animals. However, the endogenous ligand for HvNotch has not been described yet.</p> <p>Results</p> <p>We have cloned a cDNA from <it>Hydra</it>, which encodes a bona-fide Notch ligand with a conserved domain structure similar to that of Jagged-like Notch ligands from other animals. <it>Hyjagged </it>mRNA is undetectable in adult <it>Hydra </it>by <it>in situ </it>hybridisation but is strongly upregulated and easily visible at the border between bud and parent shortly before bud detachment. In contrast, HyJagged protein is found in all cell types of an adult hydra, where it localises to membranes and endosomes. Co-localisation experiments showed that it is present in the same cells as HvNotch, however not always in the same membrane structures.</p> <p>Conclusions</p> <p>The putative Notch ligand HyJagged is conserved in Cnidarians. Together with HvNotch it may be involved in the formation of the parent-bud boundary in <it>Hydra</it>. Moreover, protein distribution of both, HvNotch receptor and HyJagged indicate a more widespread function for these two transmembrane proteins in the adult hydra, which may be regulated by additional factors, possibly involving endocytic pathways.</p