Small alteration - big impacts: effects of small-scale riparian forest management on host-parasite dynamics in streams.

Environmental changes and ecological disturbances can have large and unpredictable effects on parasite dynamics. Increasing human impacts on freshwater ecosystems through land use may thus modify the distribution and abundance of parasites and have cascading effects on host populations. Here we tested the effects of small-scale riparian forest management on the nematode Cystidicoloides ephemeridarum and its insect intermediate host Ephemera danica in forested streams. We assessed the impacts of harvesting riparian trees on parasite prevalence and abundance concomitantly with host densities. We also looked at upstream and downstream reaches to document potential cascading effects on unaltered stream sections mediated by aerial dispersal of adult mayfly or downstream drift of E. danica larvae. We show that host densities and parasite levels (prevalence and abundance) increased significantly following riparian tree removal. Overall, parasite densities showed a 6- to 66-fold increase in harvested reaches compared to upstream, pristine reaches. Similar effects were also clear downstream of the disturbance. Thus, despite the small extent of riparian forest alteration along the study streams, both parasite and intermediate host were strongly affected. Small-scale riparian forest management may thus have large, unforeseen impacts on some aspects of freshwater ecosystem structure and functioning that are often ignored. Generally, understanding how human perturbations influence parasites is vital when trying to predict overall impacts on ecosystem structure and functioning, and how changes in infection dynamics may further affect host species.

Oncogenic FLT3-ITD supports autophagy via ATF4 in acute myeloid leukemia.

In acute myeloid leukemia (AML), internal tandem duplication mutations in the FLT3 tyrosine kinase receptor (FLT3-ITD) account for up to 25% of cases and are associated with a poor outcome. In order to better target this AML subtype, a comprehensive view of how FLT3-ITD impacts AML cell biology is required. Here, we found that FLT3-ITD expression increased basal autophagy in AML cells, and that both pharmacological and genetic inhibition of the receptor reduced autophagy in primary AML samples and cell lines. Conditional expression of shRNAs against key autophagy proteins demonstrated that autophagy is required for AML cell proliferation in vitro and for leukemic cells survival in a mouse model of xenograft. Importantly, autophagy inhibition also overcame FLT3 inhibitor resistance both in vitro and in vivo. The transcription factor ATF4 was identified as an essential actor of FLT3-ITD-induced autophagy. Cellular levels of ATF4 were highly dependent on FLT3-ITD activity, and downregulation of ATF4 inhibited autophagy-dependent AML cell proliferation and improved overall mouse survival similarly to autophagy inhibition. These results suggest that targeting autophagy or ATF4 in patients expressing FLT3 mutations may represent a novel promising and innovative therapeutic strategy for AML.

CyclinD-CDK4/6 complexes phosphorylate CDC25A and regulate its stability.

The phosphatase CDC25A is a key regulator of cell cycle progression by dephosphorylating and activating cyclin-CDK complexes. CDC25A is an unstable protein expressed from G1 until mitosis. CDC25A overexpression, which can be caused by stabilization of the protein, accelerates the G1/S and G2/M transitions, leading to genomic instability and promoting tumorigenesis. Thus, controlling CDC25A protein levels by regulating its stability is a critical mechanism for timing cell cycle progression and to maintain genomic integrity. Herein, we show that CDC25A is phosphorylated on Ser40 throughout the cell cycle and that this phosphorylation is established during the progression from G1 to S phase. We demonstrate that CyclinD-CDK4/CDK6 complexes mediate the phosphorylation of CDC25A on Ser40 during G1 and that these complexes directly phosphorylate this residue in vitro. Importantly, we also find that CyclinD1-CDK4 decreases CDC25A stability in a ßTrCP-dependent manner and that Ser40 and Ser88 phosphorylations contribute to this regulation. Thus our results identify cyclinD-CDK4/6 complexes as novel regulators of CDC25A stability during G1 phase, generating a negative feedback loop allowing control of the G1/S transition.

p27Kip1 represses the Pitx2-mediated expression of p21Cip1 and regulates DNA replication during cell cycle progression.

The tumor suppressor p21 regulates cell cycle progression and peaks at mid/late G1. However, the mechanisms regulating its expression during cell cycle are poorly understood. We found that embryonic fibroblasts from p27 null mice at early passages progress slowly through the cell cycle. These cells present an elevated basal expression of p21 suggesting that p27 participates to its repression. Mechanistically, we found that p27 represses the expression of Pitx2 (an activator of p21 expression) by associating with the ASE-regulatory region of this gene together with an E2F4 repressive complex. Furthermore, we found that Pitx2 binds to the p21 promoter and induces its transcription. Finally, silencing Pitx2 or p21 in proliferating cells accelerates DNA replication and cell cycle progression. Collectively, these results demonstrate an unprecedented connection between p27, Pitx2 and p21 relevant for the regulation of cell cycle progression and cancer and for understanding human pathologies associated with p27 germline mutations.

Refining the phenotypical and mutational spectrum of Taybi-Linder syndrome.

Taybi-Linder syndrome (TALS, OMIM 210710) is a rare autosomal recessive disorder belonging to the group of microcephalic osteodysplastic primordial dwarfisms (MOPD). This syndrome is characterized by short stature, skeletal anomalies, severe microcephaly with brain malformations and facial dysmorphism, and is caused by mutations in RNU4ATAC. RNU4ATAC is transcribed into a non-coding small nuclear RNA which is a critical component of the minor spliceosome. We report here four foetuses and four unrelated patients with RNU4ATAC mutations. We provide antenatal descriptions of this rare syndrome including unusual features found in two twin foetuses with compound heterozygosity for two rare mutations who presented with mild intrauterine growth retardation and atypical dysmorphic facial features. We also carried out a literature review of the patients described up to now with RNU4ATAC mutations, affected either with TALS or Roifman syndrome, a recently described allelic disorder.

p27Kip1 represses transcription by direct interaction with p130/E2F4 at the promoters of target genes.

The cyclin-cdk (cyclin-dependent kinase) inhibitor p27Kip1 (p27) has a crucial negative role on cell cycle progression. In addition to its classical role as a cyclin-cdk inhibitor, it also performs cyclin-cdk-independent functions as the regulation of cytoskeleton rearrangements and cell motility. p27 deficiency has been associated with tumor aggressiveness and poor clinical outcome, although the mechanisms underlying this participation still remain elusive. We report here a new cellular function of p27 as a transcriptional regulator in association with p130/E2F4 complexes that could be relevant for tumorigenesis. We observed that p27 associates with specific promoters of genes involved in important cellular functions as processing and splicing of RNA, mitochondrial organization and respiration, translation and cell cycle. On these promoters p27 co-localizes with p130, E2F4 and co-repressors as histone deacetylases (HDACs) and mSIN3A. p27 co-immunoprecipitates with these proteins and by affinity chromatography, we demonstrated a direct interaction of p27 with p130 and E2F4 through its carboxyl-half. We have also shown that p130 recruits p27 on the promoters, and there p27 is needed for the subsequent recruitment of HDACs and mSIN3A. Expression microarrays and luciferase assays revealed that p27 behaves as transcriptional repressor of these p27-target genes (p27-TGs). Finally, in human tumors, we established a correlation with overexpression of p27-TGs and poor survival. Thus, this new function of p27 as a transcriptional repressor could have a role in the major aggressiveness of tumors with low levels of p27.

Cytoplasmic p27 is oncogenic and cooperates with Ras both in vivo and in vitro.

p27(Kip1) (p27) can have opposing roles during malignant transformation depending on cellular context: on one hand it functions as a tumor suppressor by inhibiting cyclin-cyclin-dependent kinase (CDK) activity in the nucleus and on the other it may adopt an oncogenic role that is less well understood. To gain further insight into the roles played by p27 during tumorigenesis, we compared the susceptibility with urethane-induced tumorigenesis of two p27 mouse models, p27(-/-) and p27(CK-) knockin, in which p27 cannot bind or inhibit cyclin-CDKs. In this K-Ras-driven tumorigenesis model, p27(CK-) mice had an increase in both tumor number and aggressiveness compared with p27(-/-), indicating a cooperation between p27(CK-) and activated Ras. In the lung, increased tumorigenesis was associated with cytoplasmic localization of p27(CK-) and bronchiolaveolar stem cell amplification. The ability of p27(CK-) to cooperate with other oncogenes was not universal. When c-Myc was used as a transforming agent, p27 status became irrelevant and c-Myc was equally potent in transforming p27(+/+), p27(-/-) and p27(CK-) cells. In fact, c-Myc induced the degradation of wild-type p27 via the Skp-Cullin-F-box (SCF)-Skp2 pathway. In contrast, p27(CK-) levels were not affected by c-Myc expression, as p27(CK-) is insensitive to Skp2-mediated degradation because of its inability to bind cyclin E/CDK2. However, in presence of c-Myc, p27(CK-) remained mostly nuclear, providing an explanation for its inability to cooperate with Myc during transformation. Thus, we propose that the p27(CK-) protein needs to be localized in the cytoplasm in order to function as an oncogene, otherwise it just behaves similar to a null allele.

Surface recombination velocity measurements of efficiently passivated gold-catalyzed silicon nanowires by a new optical method.

The past decade has seen the explosion of experimental results on nanowires grown by catalyzed mechanisms. However, few are known on their electronic properties especially the influence of surfaces and catalysts. We demonstrate by an optical method how a curious electron-hole thermodynamic phase can help to characterize volume and surface recombination rates of silicon nanowires (SiNWs). By studying the electron-hole liquid dynamics as a function of the spatial confinement, we directly measured these two key parameters. We measured a surface recombination velocity of passivated SiNWs of 20 cm s(-1), 100 times lower than previous values reported. Furthermore, the volume recombination rate of gold-catalyzed SiNWs is found to be similar to that of a high-quality three-dimensional silicon crystal; the influence of the catalyst is negligible. These results advance the knowledge of SiNW surface passivation and provide essential guidance to the development of efficient nanowire-based devices.

Recombination dynamics of spatially confined electron-hole system in luminescent gold catalyzed silicon nanowires.

We study by time-resolved low temperature photoluminescence (PL) experiments of the electronic states of silicon nanowires (SiNWs) grown by gold catalyzed chemical vapor deposition and passivated by thermal SiO(2). The typical recombination line of free carriers in gold-catalyzed SiNWs (Au-SiNWs) is identified and studied by time-resolved experiments. We demonstrate that intrinsic Auger recombination governs the recombination dynamic of the dense e-h plasma generated inside the NW. In a few tens of nanoseconds after the pulsed excitation, the density of the initial electronic system rapidly decreases down to reach that of a stable electron-hole liquid phase. The comparison of the PL intensity decay time of Au-SiNWs with high crystalline quality and purity silicon layer allows us to conclude that the Au-SiNW electronic properties are highly comparable to those of bulk silicon crystal.

Direct electron imaging in electron microscopy with monolithic active pixel sensors.

A new imaging device for dynamic electron microscopy is in great demand. The detector should provide the experimenter with images having sufficient spatial resolution at high speed. Immunity to radiation damage, accumulated during exposures, is critical. Photographic film, a traditional medium, is not adequate for studies that require large volumes of data or rapid recording and charge coupled device (CCD) cameras have limited resolution, due to phosphor screen coupling. CCD chips are not suitable for direct recording due to their extreme sensitivity to radiation damage. This paper discusses characterization of monolithic active pixel sensors (MAPS) in a scanning electron microscope (SEM) as well as in a transmission electron microscope (TEM). The tested devices were two versions of the MIMOSA V (MV) chip. This 1M pixel device features pixel size of 17 x 17 microm(2) and was designed in a 0.6 microm CMOS process. The active layer for detection is a thin (less than 20 microm) epitaxial layer, limiting the broadening of the electron beam. The first version of the detector was a standard imager with electronics, passivation and interconnection layers on top of the active region; the second one was bottom-thinned, reaching the epitaxial layer from the bottom. The electron energies used range from a few keV to 30 keV for SEM and from 40 to 400 keV for TEM. Deterioration of the image resolution due to backscattering was quantified for different energies and both detector versions.

[p27Kip1 independently promotes neuronal differentiation and migration in the cerebral cortex]. / p27Kip1 independently promotes neuronal differentiation and migration in the cerebral cortex.

The generation of glutamatergic neurons by stem and progenitor cells is a complex process involving the tight coordination of multiple cellular activities, including cell cycle exit, initiation of neuronal differentiation and cell migration. The mechanisms that integrate these different events into a coherent program are not well understood. Here we show that the cyclin-dependent kinase inhibitor p27Kip1 plays an important role in neurogenesis in the mouse cerebral cortex, by promoting the differentiation and radial migration of cortical projection neurons. Importantly, p27Kip1 promotes neuronal differentiation and neuronal migration via two distinct mechanisms, which are themselves independent of the cell cycle regulatory function of p27Kip1. p27Kip1 inactivation by gene targeting or RNA interference results in neuronal differentiation and radial migration defects, demonstrating that p27Kip1 regulates cell migration in vivo. The differentiation defect, but not the migration defect, is rescued by overexpression of the proneural gene Neurogenin 2. p27Kip1 acts by stabilizing Neurogenin 2 protein, an activity carried by the N-terminal half of the protein. The migration defect resulting from p27Kp1 inactivation is rescued by blocking RhoA signalling, an activity that resides in the c-terminal half of p27Kip1. Thus, p27Kip1 plays a key role in cortical development, acting as a modular protein that independently regulates and couples multiple cellular pathways contributing to neurogenesis.

Search for supersymmetry with gauge-mediated breaking in diphoton events at D0.

We report the results of a search for supersymmetry (SUSY) with gauge-mediated breaking in the missing transverse energy distribution of inclusive diphoton events using 263 pb(-1) of data collected by the D0 experiment at the Fermilab Tevatron Collider in 2002-2004. No excess is observed above the background expected from standard model processes, and lower limits on the masses of the lightest neutralino and chargino of about 108 and 195 GeV, respectively, are set at the 95% confidence level. These are the most stringent limits to date for models with gauge-mediated SUSY breaking with a short-lived neutralino as the next-to-lightest SUSY particle.

Search for doubly charged higgs boson pair production in the decay to mu(+)mu(+)mu(-)mu(-) in pp collisions at sqrt[s]=1.96 TeV.

A search for pair production of doubly charged Higgs bosons in the process pp -->H(++)H(--) -->mu(+)mu(+)mu(-)mu(-) is performed with the D0 run II detector at the Fermilab Tevatron. The analysis is based on a sample of inclusive dimuon data collected at an energy of sqrt[s]=1.96 TeV, corresponding to an integrated luminosity of 113 pb(-1). In the absence of a signal, 95% confidence level mass limits of M(H(+/-+/-)(L))>118.4 GeV/c(2) and M(H(+/-+/-)(R))>98.2 GeV/c(2) are set for left-handed and right-handed doubly charged Higgs bosons, respectively, assuming 100% branching into muon pairs.

Observation and properties of the X(3872) decaying to J/psipi(+)pi(-) in pp collisions at sqrt[s]=1.96 TeV.

We report the observation of the X(3872) in the J/psipi(+)pi(-) channel, with J/psi decaying to mu(+)mu(-), in pp collisions at sqrt[s]=1.96 TeV. Using approximately 230 pb(-1) of data collected with the Run II D0 detector, we observe 522+/-100 X(3872) candidates. The mass difference between the X(3872) state and the J/psi is measured to be 774.9+/-3.1(stat)+/-3.0(syst) MeV/c(2). We have investigated the production and decay characteristics of the X(3872) and find them to be similar to those of the psi(2S) state.

Search for narrow tt resonances in pp collisions at square root of (s)=1.8 TeV.

A search for narrow resonances that decay into tt pairs has been performed using 130 pb(-1) of data in the lepton + jets channel collected by the Dphi detector in pp collisions at square root of (s)=1.8 TeV. There is no significant deviation observed from the standard-model predictions at a top-quark mass of 175 GeV/c2. We therefore present upper limits at the 95% confidence level on the product of the production cross section and branching fraction to tt for narrow resonances as a function of the resonance mass MX. These limits are used to exclude the existence of a leptophobic top-color particle with mass MX<560 GeV/c2, using a theoretical cross section for a width GammaX=0.012MX.

A precision measurement of the mass of the top quark.

The standard model of particle physics contains parameters--such as particle masses--whose origins are still unknown and which cannot be predicted, but whose values are constrained through their interactions. In particular, the masses of the top quark (M(t)) and W boson (M(W)) constrain the mass of the long-hypothesized, but thus far not observed, Higgs boson. A precise measurement of M(t) can therefore indicate where to look for the Higgs, and indeed whether the hypothesis of a standard model Higgs is consistent with experimental data. As top quarks are produced in pairs and decay in only about 10(-24) s into various final states, reconstructing their masses from their decay products is very challenging. Here we report a technique that extracts more information from each top-quark event and yields a greatly improved precision (of +/- 5.3 GeV/c2) when compared to previous measurements. When our new result is combined with our published measurement in a complementary decay mode and with the only other measurements available, the new world average for M(t) becomes 178.0 +/- 4.3 GeV/c2. As a result, the most likely Higgs mass increases from the experimentally excluded value of 96 to 117 GeV/c2, which is beyond current experimental sensitivity. The upper limit on the Higgs mass at the 95% confidence level is raised from 219 to 251 GeV/c2.

Search for large extra dimensions in the monojet+E(T) channel with the DØ detector.

We present a search for large extra dimensions (ED) in pp collisions at a center-of-mass energy of 1.8 TeV using data collected by the DØ detector at the Fermilab Tevatron in 1994-1996. Data corresponding to 78.8+/-3.9 pb(-1) are examined for events with large missing transverse energy, one high-p(T) jet, and no isolated muons. There is no excess observed beyond expectation from the standard model, and we place lower limits on the fundamental Planck scale of 1.0 and 0.6 TeV for 2 and 7 ED, respectively.

Search for the production of single sleptons through R-parity violation in pp; collisions at square root (s) =1.8 TeV.

We report the first search for supersymmetric particles via s-channel production and decay of smuons or muon sneutrinos at hadronic colliders. The data for the two-muon and two-jets final states were collected by the D0 experiment and correspond to an integrated luminosity of 94+/-5 pb(-1). Assuming that R parity is violated via the single coupling lambda'211, the number of candidate events is in agreement with expectation from the standard model. Exclusion contours are given in the (m(0),m(1/2)) and (m(x),m(v)) planes for lambda(')(211)=0.09, 0.08, and 0.07.