The Cohesin loading factor NIPBL recruits histone deacetylases to mediate local chromatin modifications

Cornelia de Lange Syndrome (CdLS) is a rare congenital malformation disorder. About half of the patients with CdLS carry mutations in the NIPBL gene encoding the NIPBL protein, a subunit of the Cohesin loading complex. Recent studies show association of Cohesin with chromatin-remodeling complexes, either by establishing cohesion or by recruiting Cohesin to specific chromosome locations. In yeast two-hybrid assays, we identified an interaction of NIPBL with the histone deacetylases -1 and -3. These interactions were confirmed in mammalian cells by coimmunoprecipitation and a critical region for interaction was defined to a stretch of 163 amino acids of a highly conserved region of NIPBL, which is mutated in patients with CdLS. Utilizing reporter gene assays, we could show that NIPBL fused to the GAL4-DNA-binding domain (GAL4-DBD) represses promoter activity via the recruitment of histone deacetylases. Interestingly, this effect is dramatically reduced by both NIPBL missense mutations identified in CdLS and by chemical inhibition of the histone deacetylases. Our data are the first to indicate a molecular and functional connection of NIPBL with chromatin-remodeling processes via the direct interaction with histone deacetylases

A short-term in vivo model for giant cell tumor of bone

<p>Abstract</p> <p>Background</p> <p>Because of the lack of suitable <it>in vivo </it>models of giant cell tumor of bone (GCT), little is known about its underlying fundamental pro-tumoral events, such as tumor growth, invasion, angiogenesis and metastasis. There is no existing cell line that contains all the cell and tissue tumor components of GCT and thus <it>in vitro </it>testing of anti-tumor agents on GCT is not possible. In this study we have characterized a new method of growing a GCT tumor on a chick chorio-allantoic membrane (CAM) for this purpose.</p> <p>Methods</p> <p>Fresh tumor tissue was obtained from 10 patients and homogenized. The suspension was grafted onto the CAM at day 10 of development. The growth process was monitored by daily observation and photo documentation using <it>in vivo </it>biomicroscopy. After 6 days, samples were fixed and further analyzed using standard histology (hematoxylin and eosin stains), Ki67 staining and fluorescence <it>in situ </it>hybridization (FISH).</p> <p>Results</p> <p>The suspension of all 10 patients formed solid tumors when grafted on the CAM. <it>In vivo </it>microscopy and standard histology revealed a rich vascularization of the tumors. The tumors were composed of the typical components of GCT, including (CD51+/CD68+) multinucleated giant cells whichwere generally less numerous and contained fewer nuclei than in the original tumors. Ki67 staining revealed a very low proliferation rate. The FISH demonstrated that the tumors were composed of human cells interspersed with chick-derived capillaries.</p> <p>Conclusions</p> <p>A reliable protocol for grafting of human GCT onto the chick chorio-allantoic membrane is established. This is the first <it>in vivo </it>model for giant cell tumors of bone which opens new perspectives to study this disease and to test new therapeutical agents.</p

Archeologisch bureauonderzoek en inventariserend veldonderzoek, verkennende fase. Doorn - Jan Steenlaan 15. Gemeente Utrechtse Heuvelrug (UT).

In februari 2016 is een archeologisch bureauonderzoek uitgevoerd voor het perceel aan de Jan Steenlaan 15 in Doorn (gemeente Utrechtse Heuvelrug). De aanleiding voor het onderzoek is het voornemen om de huidige opstallen te slopen en een nieuwe woning te bouwen. Hiervoor moet een omgevingsprocedure voor worden doorlopen. In dit kader is vanuit het bestaande bestemmingsplan archeologisch onderzoek noodzakelijk. Onderhavig onderzoek geeft een eerste aanzet tot een invulling van deze verplichting. Uit het bureau-onderzoek en het verkennend booronderzoek blijkt dat het plangebied in een dekzandgebied ligt, waar sinds de middeleeuwen op- en overstuiving heeft plaatsgevonden. Uit het veldonderzoek i gebleken dat het oorspronkelijke, in het dekzand aanwezige bodemprofiel geheel is verdwenen, enerzijds door overstuiving en anderzijds door bosbouw of egalisatie. Doordat het oorspronkelijke bodemprofiel is afgetopt, is de archeologische verwachting voor het plangebied laag

Archeologisch bureauonderzoek en inventariserend veldonderzoek, verkennende fase. Doorn - Jan Steenlaan 15. Gemeente Utrechtse Heuvelrug (UT).

In februari 2016 is een archeologisch bureauonderzoek uitgevoerd voor het perceel aan de Jan Steenlaan 15 in Doorn (gemeente Utrechtse Heuvelrug). De aanleiding voor het onderzoek is het voornemen om de huidige opstallen te slopen en een nieuwe woning te bouwen. Hiervoor moet een omgevingsprocedure voor worden doorlopen. In dit kader is vanuit het bestaande bestemmingsplan archeologisch onderzoek noodzakelijk. Onderhavig onderzoek geeft een eerste aanzet tot een invulling van deze verplichting. Uit het bureau-onderzoek en het verkennend booronderzoek blijkt dat het plangebied in een dekzandgebied ligt, waar sinds de middeleeuwen op- en overstuiving heeft plaatsgevonden. Uit het veldonderzoek i gebleken dat het oorspronkelijke, in het dekzand aanwezige bodemprofiel geheel is verdwenen, enerzijds door overstuiving en anderzijds door bosbouw of egalisatie. Doordat het oorspronkelijke bodemprofiel is afgetopt, is de archeologische verwachting voor het plangebied laag

Survival protein anoctamin-6 controls multiple platelet responses including phospholipid scrambling and swelling

The Scott syndrome is characterized as a mild bleeding disorder associated with a low prothrombin consumption in blood. Platelets from Scott patients show a defect in Ca2+-induced phosphatidylserine (PS) exposure on the platelet surface and microparticle formation, but unchanged Ca2+ signaling and aggregation (1, 2). For long it has been recognized that the defective PS exposure in blood cells from Scott patients results from impaired phospholipid scrambling, a process that normally abolishes the asymmetric distribution of PS and phosphatidylethanolamine over the plasma membrane upon persistent elevation of cytosolic Ca2+. Consequence of the defective PS exposure is a markedly impaired procoagulant activity of platelets, which agrees with the bleeding phenotype (3). Recently, in two unrelated Scott syndrome patients, dysfunctional mutations have been identified in the ANO6 gene (alternatively named TMEM16F), which codes for the integral membrane protein anoctamin 6 (ANO6) (4, 5). The question can be raised if a gene defect in ANO6 alone is sufficient for the altered blood cell properties in the Scott syndrome. In the present article, we used several molecular and functional approaches to unravel the apparently multiple and non-redundant functions of ANO6 in platelets. We investigated the alterations in platelet properties of a Scott patient in comparison to platelets from healthy control subjects, and furthermore compared the platelet properties of several new strains of Ano6-deficient mice versus wild type mice

Survival protein anoctamin-6 controls multiple platelet responses including phospholipid scrambling and swelling

The Scott syndrome is characterized as a mild bleeding disorder associated with a low prothrombin consumption in blood. Platelets from Scott patients show a defect in Ca2+-induced phosphatidylserine (PS) exposure on the platelet surface and microparticle formation, but unchanged Ca2+ signaling and aggregation (1, 2). For long it has been recognized that the defective PS exposure in blood cells from Scott patients results from impaired phospholipid scrambling, a process that normally abolishes the asymmetric distribution of PS and phosphatidylethanolamine over the plasma membrane upon persistent elevation of cytosolic Ca2+. Consequence of the defective PS exposure is a markedly impaired procoagulant activity of platelets, which agrees with the bleeding phenotype (3). Recently, in two unrelated Scott syndrome patients, dysfunctional mutations have been identified in the ANO6 gene (alternatively named TMEM16F), which codes for the integral membrane protein anoctamin 6 (ANO6) (4, 5). The question can be raised if a gene defect in ANO6 alone is sufficient for the altered blood cell properties in the Scott syndrome. In the present article, we used several molecular and functional approaches to unravel the apparently multiple and non-redundant functions of ANO6 in platelets. We investigated the alterations in platelet properties of a Scott patient in comparison to platelets from healthy control subjects, and furthermore compared the platelet properties of several new strains of Ano6-deficient mice versus wild type mice