The binding of copper in human ceruloplasmin

Zusammenfassung Mit Photooxydation und Diazotierung wird gezeigt, dass Histidinreste an der Bindung von Kupfer an Caeruloplasmin beteiligt sind.     

Improving national-scale invasion maps: tamarisk in the western United States

New invasions, better field data, and novel spatial-modeling techniques often drive the need to revisit previous maps and models of invasive species. Such is the case with the at least 10 species of Tamarix, which are invading riparian systems in the western United States and expanding their range throughout North America. In 2006, we developed a National Tamarisk Map by using a compilation of presence and absence locations with remotely sensed data and statistical modeling techniques. Since the publication of that work, our database of Tamarix distributions has grown significantly. Using the updated database of species occurrence, new predictor variables, and the maximum entropy (Maxent) model, we have revised our potential Tamarix distribution map for the western United States. Distance-to-water was the strongest predictor in the model (58.1%), while mean temperature of the warmest quarter was the second best predictor (18.4%). Model validation, averaged from 25 model iterations, indicated that our analysis had strong predictive performance (AUC = 0.93) and that the extent of Tamarix distributions is much greater than previously thought. The southwestern United States had the greatest suitable habitat, and this result differed from the 2006 model. Our work highlights the utility of iterative modeling for invasive species habitat modeling as new information becomes available. A menudo las nuevas invasiones, mejores datos de campo y técnicas novedosas de modelado espacial impulsan la actualización de los mapas y de los modelos existentes de especies invasoras. Este es el caso de al menos 10 especies de Tamarix, las cuales están invadiendo los sistemas ribereños en el oeste de los EE.UU. y extendiendo su distribución por toda Norteamérica. En 2006, desarrollamos un mapa nacional del tamarisco (National Tamarisk Map) utilizando una compilación de sitios de presencia y ausencia con datos de sensores remotos y técnicas de modelación estadística. Desde la publicación de este trabajo, nuestra base de datos sobre la distribución de Tamarix ha crecido considerablemente. Utilizando la base de datos actualizada de presencia de especies, nuevas variables predictoras y el modelo de máxima entropía (Maxent), hemos modificado nuestro mapa de la distribución potencial de Tamarix para el oeste de los EE.UU. El predictor más fuerte en el modelo fue la distancia al agua (58.1%), y la temperatura promedio del trimestre más cálido fue el segundo (18.4%). La validación de modelo, calculada como el promedio de 25 iteraciones del modelo, indicó que nuestro análisis tuvo una alta capacidad predictiva (ABC = 0.93), y que la distribución de Tamarix es mucho más extensa de lo que se pensaba. El suroeste de los EE.UU. tuvo la mayor cantidad de hábitat adecuado para la especie, y este resultado difirió del modelo de 2006. Nuestro trabajo enfatiza la utilidad del modelado iterativo para modelar el hábitat de las especies invasoras a medida que se disponga de nueva información.     

HepaticΔ 4-steroid hydrogenase in the pregnant rat

Résumé Chez la rate gravide, la réduction du corticosterone, exprimé par gramme de foie fut augmentée les douzièmes et quatorzième jours de gestation. L'activité du foie entier augmenta à partir du douzième jour.     

IgH class switching and translocations use a robust non-classical end-joining pathway

Immunoglobulin variable region exons are assembled in developing B cells by V(D)J recombination. Once mature, these cells undergo class-switch recombination (CSR) when activated by antigen. CSR changes the heavy chain constant region exons (Ch) expressed with a given variable region exon from Cmu to a downstream Ch (for example, Cgamma, Cepsilon or Calpha), thereby switching expression from IgM to IgG, IgE or IgA. Both V(D)J recombination and CSR involve the introduction of DNA double-strand breaks and their repair by means of end joining. For CSR, double-strand breaks are introduced into switch regions that flank Cmu and a downstream Ch, followed by fusion of the broken switch regions. In mammalian cells, the 'classical' non-homologous end joining (C-NHEJ) pathway repairs both general DNA double-strand breaks and programmed double-strand breaks generated by V(D)J recombination. C-NHEJ, as observed during V(D)J recombination, joins ends that lack homology to form 'direct' joins, and also joins ends with several base-pair homologies to form microhomology joins. CSR joins also display direct and microhomology joins, and CSR has been suggested to use C-NHEJ. Xrcc4 and DNA ligase IV (Lig4), which cooperatively catalyse the ligation step of C-NHEJ, are the most specific C-NHEJ factors; they are absolutely required for V(D)J recombination and have no known functions other than C-NHEJ. Here we assess whether C-NHEJ is also critical for CSR by assaying CSR in Xrcc4- or Lig4-deficient mouse B cells. C-NHEJ indeed catalyses CSR joins, because C-NHEJ-deficient B cells had decreased CSR and substantial levels of IgH locus (immunoglobulin heavy chain, encoded by Igh) chromosomal breaks. However, an alternative end-joining pathway, which is markedly biased towards microhomology joins, supports CSR at unexpectedly robust levels in C-NHEJ-deficient B cells. In the absence of C-NHEJ, this alternative end-joining pathway also frequently joins Igh locus breaks to other chromosomes to generate translocations.     

Thymosin beta4 induces adult epicardial progenitor mobilization and neovascularization

Cardiac failure has a principal underlying aetiology of ischaemic damage arising from vascular insufficiency. Molecules that regulate collateral growth in the ischaemic heart also regulate coronary vasculature formation during embryogenesis. Here we identify thymosin beta4 (Tbeta4) as essential for all aspects of coronary vessel development in mice, and demonstrate that Tbeta4 stimulates significant outgrowth from quiescent adult epicardial explants, restoring pluripotency and triggering differentiation of fibroblasts, smooth muscle cells and endothelial cells. Tbeta4 knockdown in the heart is accompanied by significant reduction in the pro-angiogenic cleavage product N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP). Although injection of AcSDKP was unable to rescue Tbeta4 mutant hearts, it significantly enhanced endothelial cell differentiation from adult epicardially derived precursor cells. This study identifies Tbeta4 and AcSDKP as potent stimulators of coronary vasculogenesis and angiogenesis, and reveals Tbeta4-induced adult epicardial cells as a viable source of vascular progenitors for continued renewal of regressed vessels at low basal level or sustained neovascularization following cardiac injury.     

Template switching during break-induced replication

DNA double-strand breaks (DSBs) are potentially lethal lesions that arise spontaneously during normal cellular metabolism, as a consequence of environmental genotoxins or radiation, or during programmed recombination processes. Repair of DSBs by homologous recombination generally occurs by gene conversion resulting from transfer of information from an intact donor duplex to both ends of the break site of the broken chromosome. In mitotic cells, gene conversion is rarely associated with reciprocal exchange and thus limits loss of heterozygosity for markers downstream of the site of repair and restricts potentially deleterious chromosome rearrangements. DSBs that arise by replication fork collapse or by erosion of uncapped telomeres have only one free end and are thought to repair by strand invasion into a homologous duplex DNA followed by replication to the chromosome end (break-induced replication, BIR). BIR from one of the two ends of a DSB would result in loss of heterozygosity, suggesting that BIR is suppressed when DSBs have two ends so that repair occurs by the more conservative gene conversion mechanism. Here we show that BIR can occur by several rounds of strand invasion, DNA synthesis and dissociation. We further show that chromosome rearrangements can occur during BIR if dissociation and reinvasion occur within dispersed repeated sequences. This dynamic process could function to promote gene conversion by capture of the displaced invading strand at two-ended DSBs to prevent BIR.     

A murine lung cancer co-clinical trial identifies genetic modifiers of therapeutic response

Targeted therapies have demonstrated efficacy against specific subsets of molecularly defined cancers. Although most patients with lung cancer are stratified according to a single oncogenic driver, cancers harbouring identical activating genetic mutations show large variations in their responses to the same targeted therapy. The biology underlying this heterogeneity is not well understood, and the impact of co-existing genetic mutations, especially the loss of tumour suppressors, has not been fully explored. Here we use genetically engineered mouse models to conduct a 'co-clinical' trial that mirrors an ongoing human clinical trial in patients with KRAS-mutant lung cancers. This trial aims to determine if the MEK inhibitor selumetinib (AZD6244) increases the efficacy of docetaxel, a standard of care chemotherapy. Our studies demonstrate that concomitant loss of either p53 (also known as Tp53) or Lkb1 (also known as Stk11), two clinically relevant tumour suppressors, markedly impaired the response of Kras-mutant cancers to docetaxel monotherapy. We observed that the addition of selumetinib provided substantial benefit for mice with lung cancer caused by Kras and Kras and p53 mutations, but mice with Kras and Lkb1 mutations had primary resistance to this combination therapy. Pharmacodynamic studies, including positron-emission tomography (PET) and computed tomography (CT), identified biological markers in mice and patients that provide a rationale for the differential efficacy of these therapies in the different genotypes. These co-clinical results identify predictive genetic biomarkers that should be validated by interrogating samples from patients enrolled on the concurrent clinical trial. These studies also highlight the rationale for synchronous co-clinical trials, not only to anticipate the results of ongoing human clinical trials, but also to generate clinically relevant hypotheses that can inform the analysis and design of human studies.