73 research outputs found
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The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis
The pathogenesis of primary sclerosing cholangitis (PSC) and the mechanistic link to inflammatory bowel disease remain illâdefined. Ectonucleoside triphosphate diphosphohydrolaseâ1 (ENTPD1)/clusters of differentiation (CD) 39, the dominant purinergic ectoâenzyme, modulates intestinal inflammation. Here, we have explored the role of CD39 in biliary injury and fibrosis. The impact of CD39 deletion on disease severity was studied in multidrug resistance protein 2 (Mdr2)â/â and 3,5âdiethoxycarbonylâ1,4âdihydrocollidine mouse models of sclerosing cholangitis and biliary fibrosis. Antibodyâmediated CD8+ Tâcell depletion, selective gut decontamination, experimental colitis, and administration of stable adenosine triphosphate (ATP) agonist were performed. Retinoic acidâinduced gut imprinting on T cells was studied in vitro. Over half of Mdr2â/â;CD39â/â double mutants, expected by Mendelian genetics, died in utero. Compared to Mdr2â/â;CD39+/+, surviving Mdr2â/â;CD39â/â mice demonstrated exacerbated liver injury, fibrosis, and ductular reaction. CD39 deficiency led to a selective increase in hepatic CD8+ T cells and integrin α4ÎČ7, a Tâcell gutâtropism receptor. CD8+ cell depletion in Mdr2â/â;CD39â/â mice diminished hepatobiliary injury and fibrosis. Treatment with antibiotics attenuated, whereas dextran sulfate sodiumâinduced colitis exacerbated, liver fibrosis in Mdr2â/â mice. Colonic administration of αÎČâATP into CD39âsufficient Mdr2â/â mice triggered hepatic CD8+ cell influx and recapitulated the severe phenotype observed in Mdr2â/â;CD39â/â mice. In vitro, addition of ATP promoted the retinoic acidâinduced imprinting of gutâhoming integrin α4ÎČ7 on naive CD8+ cells. CD39 expression was relatively low in human normal or PSC livers but abundantly present on immune cells of the colon and further upâregulated in samples of patients with inflammatory bowel disease. Conclusion:: CD39 deletion promotes biliary injury and fibrosis through gutâimprinted CD8+ T cells. Pharmacological modulation of purinergic signaling may represent a promising approach for the treatment of PSC. (Hepatology Communications 2017;1:957â972
Enabling large-scale design, synthesis and validation of small molecule protein-protein antagonists
Although there is no shortage of potential drug targets, there are only a handful known low-molecular-weight inhibitors of protein-protein interactions (PPIs). One problem is that current efforts are dominated by low-yield high-throughput screening, whose rigid framework is not suitable for the diverse chemotypes present in PPIs. Here, we developed a novel pharmacophore-based interactive screening technology that builds on the role anchor residues, or deeply buried hot spots, have in PPIs, and redesigns these entry points with anchor-biased virtual multicomponent reactions, delivering tens of millions of readily synthesizable novel compounds. Application of this approach to the MDM2/p53 cancer target led to high hit rates, resulting in a large and diverse set of confirmed inhibitors, and co-crystal structures validate the designed compounds. Our unique open-access technology promises to expand chemical space and the exploration of the human interactome by leveraging in-house small-scale assays and user-friendly chemistry to rationally design ligands for PPIs with known structure. © 2012 Koes et al
Acquired resistance to oxaliplatin is not directly associated with increased resistance to DNA damage in SK-N-ASrOXALI4000, a newly established oxaliplatin-resistant sub-line of the neuroblastoma cell line SK-N-AS
The formation of acquired drug resistance is a major reason for the failure of anti-cancer therapies after initial response. Here, we introduce a novel model of acquired oxaliplatin resistance, a sub-line of the non-MYCN-amplified neuroblastoma cell line SK-N-AS that was adapted to growth in the presence of 4000 ng/mL oxaliplatin (SK-N-ASrOXALI4000). SK-N-ASrOXALI4000 cells displayed enhanced chromosomal aberrations compared to SK-N-AS, as indicated by 24-chromosome fluorescence in situ hybridisation. Moreover, SK-N-ASrOXALI4000 cells were resistant not only to oxaliplatin but also to the two other commonly used anti-cancer platinum agents cisplatin and carboplatin. SK-N-ASrOXALI4000 cells exhibited a stable resistance phenotype that was not affected by culturing the cells for 10 weeks in the absence of oxaliplatin. Interestingly, SK-N-ASrOXALI4000 cells showed no cross resistance to gemcitabine and increased sensitivity to doxorubicin and UVC radiation, alternative treatments that like platinum drugs target DNA integrity. Notably, UVC-induced DNA damage is thought to be predominantly repaired by nucleotide excision repair and nucleotide excision repair has been described as the main oxaliplatin-induced DNA damage repair system. SK-N-ASrOXALI4000 cells were also more sensitive to lysis by influenza A virus, a candidate for oncolytic therapy, than SK-N-AS cells. In conclusion, we introduce a novel oxaliplatin resistance model. The oxaliplatin resistance mechanisms in SK-N-ASrOXALI4000 cells appear to be complex and not to directly depend on enhanced DNA repair capacity. Models of oxaliplatin resistance are of particular relevance since research on platinum drugs has so far predominantly focused on cisplatin and carboplatin
Reproduktion fundamentaler menschlicher Miïżœbildungen am Hïżœhnchenkeim durch Sauerstoffmangel
Identification of organic arsenic, tin, antimony and tellurium compounds in environmental samples by GC-MS
In this study, the potential of GC-MS for structure analysis of organometal(loid) species of arsenic, tin, antimony and tellurium in environmental samples was investigated. Analytes were ionic organometal(loid) species from compost samples, which were derivatised with NaBH4 before introduction into the GC/MS via a purge and trap system, as well as volatile species from hydrothermal gases sampled on SPME-fibres. Several organic species of arsenic, tin, antimony and tellurium were identified. By scanning the chromatogram for typical fragments of organoarsenic compounds seven arsenic species were identified. To our knowledge three of them (diarsine, monomethyl diarsine and dimethylarsenomercaptane) have not been reported in environmental samples so far. To further exclude artefacts being misinterpreted as volatile organometal(loid)s, measurements of volatile arsenic species naturally occurring in hydrothermal samples with SPME-GC-MS were performed. The determination of dimethylarsenomercaptane in these samples confirms that this compound which was also found after derivatisation of ionic arsenic species is not an artefact. A strategy for the identification of unknown species whose structure cannot be elucidated by library comparison or with the help of isotopic patterns is presented. The necessity of positive identification of unknown species and confirmation of known and assumed compounds for speciation analysis is highlighted and the applicability of quadrupol GC/MS for this purpose is stated
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