Identification and characterisation of mutations associated with von Willebrand disease in a Turkish patient cohort

Several cohort studies have investigated the molecular basis of von Willebrand disease (VWD); however, these have mostly focused on European and North American populations. This study aimed to investigate mutation spectrum in 26 index cases (IC) from Turkey diagnosed with all three VWD types, the majority (73%) with parents who were knowingly related. IC were screened for mutations using multiplex ligation-dependent probe amplification and analysis of all von Willebrand factor gene (VWF) exons and exon/intron boundaries. Selected missense mutations were expressed in vitro. Candidate VWF mutations were identified in 25 of 26 IC and included propeptide missense mutations in four IC (two resulting in type 1 and two in recessive 2A), all influencing VWF expression in vitro. Four missense mutations, a nonsense mutation and a small in-frame insertion resulting in type 2A were also identified. Of 15 type 3 VWD IC, 13 were homozygous and two compound heterozygous for 14 candidate mutations predicted to result in lack of expression and two propeptide missense changes. Identification of intronic breakpoints of an exon 17–18 deletion suggested that the mutation resulted from non-homologous end joining. This study provides further insight into the pathogenesis of VWD in a population with a high degree of consanguineous partnerships

The histone methyltransferase Setd8 acts in concert with c-Myc and is required to maintain skin

Keratinocyte-specific ablation of the histone H4K20 methyltransferase Setd8 reveals its essential role in embryonic and postnatal skin homeostasis. Molecularly, the c-myc target gene Setd8 regulates proliferation/differentiation by controlling p63 function

Effects of antidepressants on K_2P-channels in neuronal and cardiac cells

Die vorliegende Arbeit beschäftigte sich mit der Wirkung von Antidepressiva auf K2P-Kanäle. Sie stellen wie spannungsabhängige Ca2+, Na+ und K+-Kanäle als neuronale Ionenkanäle aufgrund ihrer Expressionsmuster und physiologischen Eigenschaften potentielle Zielproteine für Antidepressiva dar. Darum werden K2P-Kanäle in heterologen Expressionssystemen von klinisch verabreichten Antidepressiva inhibiert. Die K2P-Kanäle TREK-1, TASK-1 und THIK-1 zeigten sich in dieser Arbeit alle sensitiv auf das Antidepressivum Fluoxetin, welches die Kaliumströme der Kanäle unterschiedlich stark inhibierte. Hierbei lieferten die vorliegenden Untersuchungen den Nachweis, dass TREK-1 auf Fluoxetin am meisten, THIK-1 am wenigsten sensitiv reagiert. Der humane TREK-1 wird durch Fluoxetin in den Expressionssystemen Oozyten und HEK-Zellen zu fast 80% inhibiert, wobei bei der humanen Zelllinie nur ein Zehntel der vorher eingesetzten Antidepressivakonzentration für die gleiche Inhibition des Auswärtsstroms notwendig war. Die vorliegende Arbeit weist Inhibitionen des Kanals bei einer Fluoxetinkonzentration von 1 µM nach, was der Serumkonzentration von depressiven Patienten entspricht. Zudem wird TREK-1 durch die Antidepressiva Maprotilin, Mirtazapin, Citalopram, Doxepin und Venlafaxin inhibiert, wobei letzteres kaum eine Wirkung zeigt. Alle verwendeten Antidepressiva nutzen die gleichen Angriffspunkte am Kanalprotein, da es bei einer Koapplikation mit einem weiteren Antidepressivum oder Benzodiazepin zu keiner Inhibitionsverstärkung kommt. Die Interaktion zwischen Antidepressivum und Kanalprotein verläuft mit großer Wahrscheinlichkeit direkt und ohne „second-messenger-Wege“. Hierbei konnten die porenformende Region und der C-Terminus des Kanals als Interaktionspartner ausgeschlossen werden. Der Mechanismus der alternativen Translations-Initiaton generiert zwei unterschiedliche Proteinprodukte aus einem TREK-1 Transkript, eine lange Version des Proteins mit 426 Aminosäuren und zusätzlich eine kurze Version mit 374 Aminosäuren, welcher die ersten 52 N-terminalen Aminosäuren fehlen. Die Fluoxetin-Sensitivität von TREK-1 [N52] verringert sich um 70%. Dies verdeutlicht, dass die ersten 52 Aminosäuren essentiell zur TREK-1 Interaktion mit Antidepressiva beitragen.The study at hand is about the effect of antidepressants on K2P-channels. As neuronal ion-channels like voltage-gated Ca2+, Na+ and K+-channels, the K2P-channels constitute a potential target for antidepressants because of their tissue expression and physiological characteristics. Clinically prescribed antidepressants inhibit the K2P-channels in heterologous expression systems for that reason. In our experiments the K2P-channels TREK-1, TASK-1 and THIK-1 were sensitive to the antidepressant Fluoxetine, which inhibited the potassium current in different ways. The study provides evidence that TREK-1 reacts to Fluoxetine most sensitively whereas THIK-1 reacts least. The humane TREK-1 is inhibited up to 80% by Fluoxetine in expression systems oocytes and HEK-cells, in which only a tenth of the antidepressant concentration induced the same current inhibition. Our experiments showed already a channel block already at 1 µM Fluoxetine concentration, which is conform to the antidepressant serum concentration of depressive patients. Furthermore TREK-1 is inhibited by the antidepressants Maprotiline, Mirtazapine, Citalopram, Doxepin and Venlafaxine, whereas the last one showed least effects. The used antidepressants occupy the same targets at the channel protein, because a coapplication with a further antidepressant or benzodiazepine didn´t increase the maximum channel block. The interaction between antidepressant and channel protein is working directly without second messenger pathway. The pore forming region and the C-terminus of the channels could be excluded as interaction partner. Alternative translation initiation (ATI) generates two different protein products from a single transcript of TREK-1, a long version of the protein with 426 amino acids and in addition a short version with 374 amino acids, lacking the first 52 amino acids at the N-terminus. The sensitivity of TREK-1[N52] to fluoxetine declined by 70% indicating that the first 52 amino acids essentially contribute to the interaction of TREK-1 with the antidepressant

Zeitgeschehen | [Buchbesprechungen Nr. 294–299]

Axel Schildt (Hg.): Von draußen. Ausländische intellektuelle Einflüsse in der Bundesrepublik bis 1990 (Eckhard Jesse) Melanie Amann: Angst für Deutschland. Die Wahrheit über die AfD. Wo sie herkommt, wer sie führt, wohin sie steuert (Eckhard Jesse) Hans-Joachim Lauth, Marianne Kneuer, Gert Pickel (Hg.): Handbuch Vergleichende Politikwissenschaft (Eckhard Jesse) Ulrich Brand, Markus Wissen: Imperiale Lebensweise. Zur Ausbeutung von Mensch und Natur im globalen Kapitalismus (Dieter Senghaas) Peter Böthig (Hg.): sprachzeiten: Der Literarische Salon von Ekke Maaß. Eine Dokumentation von 1978-2016 (Rainer Eckert) Eckhard Jesse (Hg.): Wie gefährlich ist Extremismus? Gefahren durch Extremismus, Gefahren in Umgang mit Extremismus (Michaela Heinze

Archaea and Bacteria in deep lake hypolimnion: In situ dark inorganic carbon uptake

The interest for microorganisms inhabiting the hypolimnion and for their role in biogeochemical cycles of lakes is considerable, but knowledge is far from complete. The presence of chemolithoautotrophic Bacteria and mesophilic Archaea (e.g., Thaumarchaeota) assimilating inorganic carbon in the deep hypolimnion of lakes has been ascertained. We measured, for the first time at 350 m in Lake Maggiore (Northern Italy), the prokaryotic in situ dark [14C]HCO3 incorporation with a new custom-made apparatus, which takes samples and adds tracers in situ. Thereby stress factors affecting prokaryotes during sample recovery from the depth were avoided. We tested the new instrument at different depths and conditions, performing parallel conventional on board incubations. We found that dark [14C]HCO3 incorporations had lower standard deviation in in situ incubations with respect to the on board ones, but their means were not statistically different. At 350 m we estimated an uptake of 187.7±15 μg C m–3 d–1, which is in line with the published uptake rates in aquatic systems. By inhibiting the bacterial metabolism, we found that Archaea were responsible for 28% of the total CO2 uptake. At the same depth, Thaumarchaeota, on average, constituted 11% of total DAPI counts. Dark [14C]HCO3 incorporation integrated along the aphotic water column was 65.8±5.2 mg C m–2 d–1 which corresponds to 87% of picophytoplanktonic autotrophic fixation in the euphotic layer. This study provides the first evidence of Bacteria and Archaea dark CO2 fixation in the deep hypolimnion of a subalpine lake and indicates a potentially significant prokaryotic CO2 sink