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.

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.

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.

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.

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.

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.

Involvement of p21(Waf1/Cip1) in protein kinase C alpha-induced cell cycle progression.

Protein kinase C (PKC) plays an important role in the regulation of glioma growth; however, the identity of the specific isoform and mechanism by which PKC fulfills this function remain unknown. In this study, we demonstrate that PKC activation in glioma cells increased their progression through the cell cycle. Of the six PKC isoforms that were present in glioma cells, PKC alpha was both necessary and sufficient to promote cell cycle progression when stimulated with phorbol 12-myristate 13-acetate. Also, decreased PKC alpha expression resulted in a marked decrease in cell proliferation. The only cell cycle-regulatory molecule whose expression was rapidly altered and increased by PKC alpha activity was the cyclin-cyclin-dependent kinase (CDK) inhibitor p21(Waf1/Cip1). Coimmunoprecipitation studies revealed that p21(Waf1/Cip1) upregulation was accompanied by an incorporation of p21(Waf1/Cip1) into various cyclin-CDK complexes and that the kinase activity of these complexes was increased, thus resulting in cell cycle progression. Furthermore, depletion of p21(Waf1/Cip1) by antisense strategy attenuated the PKC-induced cell cycle progression. These results suggest that PKC alpha activity controls glioma cell cycle progression through the upregulation of p21(Waf1/Cip1), which facilitates active cyclin-CDK complex formation.

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.

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.

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.

Differential activation of ERKs to focal adhesions by PKC epsilon is required for PMA-induced adhesion and migration of human glioma cells.

Protein kinase C (PKC) is a family of serine/threonine kinases involved in the transduction of a variety of signals. There is increasing evidence to indicate that specific PKC isoforms are involved in the regulation of distinct cellular processes. In glioma cells, PKC alpha was found to be a critical regulator of proliferation and cell cycle progression, while PKC epsilon was found to regulate adhesion and migration. Herein, we report that specific PKC isoforms are able to differentially activate extracellular-signal regulated kinase (ERK) in distinct cellular locations: while PKC alpha induces the activation of nuclear ERK, PKC epsilon induces the activation of ERK at focal adhesions. Inhibition of the ERK pathway completely abolished the PKC-induced integrin-mediated adhesion and migration. Thus, we present the first evidence that PKC epsilon is able to activate ERK at focal adhesions to mediate glioma cell adhesion and motility, providing a molecular mechanism to explain the different biological functions of PKC alpha and epsilon in glioma cells.

Effects of morphine, naloxone and their interaction in the learned-helplessness paradigm in rats.

The aim of this study was to investigate the possible involvement of the mu-opioid system on the learned-helplessness paradigm, an experimental model of depression, in rats. In this test, rats were first exposed to inescapable foot-shocks (IS); 48 h later, they were submitted to a daily shuttle-box session (30 trials) for 3 consecutive days. Avoidance responses, escape failures and animal activity during each intertrial interval were recorded. Twice daily injections of morphine (0.25-8 mg/kg per day, SC), a mu-opioid agonist, reduced the increased escape failures induced by IS, as did tricyclic antidepressants. Significantly higher intertrial activity was observed in rats treated with morphine (2-8 mg/kg per day) compared with their associated control groups. Naloxone (1 and 2 mg/kg, IP), a mu-opioid antagonist, injected 10 min before each shuttle-box session impaired escape behavior in non-stressed rats and worsened the escape deficit induced by IS. Morphine-induced improvement of escape behavior and increase in intertrial activity were clearly reversed by a low inactive dose of naloxone (0.5 mg/kg). These results suggest that mu-opioid receptor mediation is involved in the deleterious effects of uncontrollable stress.

Effects of the co-administration of mirtazapine and paroxetine on serotonergic neurotransmission in the rat brain.

The alpha(2)-adrenoreceptor antagonist mirtazapine, which is also a 5-HT(2), 5-HT(3) and H(1) receptors antagonist and the selective serotonin (5-HT) reuptake inhibitor paroxetine are effective antidepressant drugs which enhance 5-HT neurotransmission via different mechanisms. The present studies were undertaken to determine whether the mirtazapine-paroxetine combination could induce an earlier and/or a greater effect on the 5-HT system than either drug alone. Using in vivo electrophysiological paradigms, the firing activity of dorsal raphe 5-HT neurons was decreased by 70% in rats treated with paroxetine (10 mg/kg/day, s.c.) for 2 days and was back to normal after 21 days. In contrast, a 2-day treatment with mirtazapine (5 mg/kg/day, s.c.) did not alter the firing of 5-HT neurons whereas it was increased by 60% after 21 days of treatment. A low dose of mirtazapine (5 mg/kg/day, s.c.x2 days) failed to offset the decremental effect of paroxetine on the 5-HT neuron firing activity, but a higher dose (10 mg/kg/day, s.c.x2 days) did attenuate the decremental effect of paroxetine. In the dorsal hippocampus, neither mirtazapine (5 mg/kg/day, s.c.) nor a paroxetine (10 mg/kg/day, s.c.) treatment altered the responsiveness of 5-HT(1A) receptors to microiontophoretically-applied 5-HT. Both in controls and in rats treated for 2 days with paroxetine alone, the administration of the 5-HT(1A) antagonist WAY 100635 (25-100 microg/kg, i.v.) did not change the firing activity of dorsal hippocampus CA(3) pyramidal neurons. However, WAY 100635 increased significantly the firing activity of these neurons in rats treated with mirtazapine alone but to a greater extent with both mirtazapine and paroxetine for 2 days. After 21 days of treatment, WAY 100635 increased to a greater degree the firing rate of CA(3) pyramidal neurons in rats which received the combination over rats given either drug alone. It is concluded that the mirtazapine-paroxetine combination shortened the delay in enhancing the tonic activation of postsynaptic 5-HT(1A) receptors and produced a greater activation of the postsynaptic 5-HT(1A) receptors than either drug given alone. The present results suggested that mirtazapine may have a faster onset of action than a SSRI, and that the co-administration of mirtazapine and paroxetine may accelerate the antidepressant response and as well as being more effective than either drug alone.

The Roux-Y loop in modern digestive tract surgery.

The technique of Y loop was utilized by César Roux beginning in January 1892 for antral or pyloric obstruction. He used the proximal jejunum to perform a Y gastroenterostomy. However, he abandoned the procedure in 1911, having noticed the frequency of late peptic ulcerations in the loop. Nevertheless, the Y-branching idea was later reconsidered and applications in hepatic, biliary, and pancreatic surgery were developed, as well as applications in gastrointestinal surgery. In this latter area, Roux-Y anastomoses are now performed primarily or secondarily. Primary uses include restoring the continuity of the alimentary tract after total gastrectomy (with or without a pouch), reconstruction after Whipple's procedure, repair of duodenal trauma, and intentional digestive bypass for morbid obesity. Secondary uses include remedial procedures after simple esophagojejunostomy, gastric fundus resection, and a Billroth II procedure to obviate postoperative complications due to bile reflux.

Reversal of learned helplessness by morphine in rats: involvement of a dopamine mediation.

The aim of this study was to examine the role of dopamine neurotransmission in the effects of morphine in the learned helplessness paradigm in rats, a generally recognized model of depression. In this model, rats first exposed to inescapable shocks (stressed rats) exhibited an escape deficit in a subsequent shuttle-box test performed 48 h later for 3 consecutive days. The numbers of escape failures and intertrial crossings (motor activity during each intertrial interval) were recorded. Morphine was injected twice daily for 5 days (6 mg/kg/day, s.c.), and haloperidol, a preferential D2-dopamine receptor antagonist, was injected i.p. 15 min before each shuttle-box session. At the highest dose tested (150 microg/kg) haloperidol mimicked the behavioral deficit produced by inescapable shocks. A 37.5 microg/kg dose of haloperidol, which was ineffective by itself, reversed the morphine-induced improvement of escape behavior in previously stressed rats and the morphine-induced increase in intertrial activity in both stressed and nonstressed animals. These results support roles (a) for a dysregulation of dopaminergic neuronal activity in the expression of escape deficit subsequent to an inescapable aversive situation, and (b) for a dopaminergic mediation in the effects of morphine in the learned helplessness paradigm.

Dopaminergic and opioidergic mediations of tricyclic antidepressants in the learned helplessness paradigm.

The roles of dopaminergic and opioid neurotransmissions in the activity of three tricyclic antidepressants endowed with different monoamine-reuptake properties [desipramine (DESI), imipramine (IMI), amineptine (AMN)] were examined using a behavioral model of depression in rats; the learned helplessness paradigm. In this model, exposure of rats to inescapable shocks (day 1) produced a subsequent escape deficit in a shuttle box test (days 3, 4, and 5). The escape deficit was reversed by AMN, DESI, and IMI administered twice daily for 5 days (16 and 32 mg/kg/day, p < 0.05, days 3, 4, and 5). In addition, AMN tended to enhance the motor activity of rats during the intertrial intervals, but on the first shuttle-box test only (day 3: p < 0.05, control vs AMN). Haloperidol, a preferential D2 dopamine receptor antagonist, acutely injected IP (37.5 microg/kg), suppressed the behavioral activity of DESI and IMI but not that of AMN. Naloxone, a preferential mu-opioid receptor antagonist, acutely injected IP (0.5 mg/kg), suppressed the behavioral activity of IMI but not that of DESI and AMN. It is concluded that an increased dopaminergic activity is a neurochemical effect common to the different tricyclic antidepressants (via a presynaptic mechanism for AMN and a postsynaptic mechanism for DESI and IMI), whereas an increased mu-opioid neurotransmission does not appear to be essential.

[Critical analysis of different methods used for toxicology screening in emergency laboratory]. / Analyse critique des différentes méthodes utilisées pour le dépistage toxicologique dans un laboratoire d'urgence.

Emergency analysis in toxicology is a difficult exercise. It involves in diagnosis, prognosis and the treatment of intoxication. Several methods exist in emergency screening. We have distinguished three large groups. Based on specificity: screening methods of medicament family (chemical methods and immunoassays) with benzodiazepines, tricyclic antidepressants, barbiturates and phenothiazines; complementary screening methods (thin layer chromatography and high pressure liquid chromatography) for a wider screening and finally quantitative methods (enzymatic, immunoassay, spectrometry and chromatography) specific to a molecule. The first group allows a rapid qualitative research according to medicament class but lacks specificity. The second group represented by the Remedi system, offers a larger screening of molecules but is more expensive and cannot detect classic molecules. The third group allows a precise dosage but is restricted to one molecule. We need one or the other of methods following clinical context and the type of molecule. In our laboratory, we have eliminated barbiturates and benzodiazepines research. We search only tricyclic antidepressants, salicylates and paracetamol. The Remedi system acts as a complement. It is essential to have a good knowledge of the limits and specificity of each method in order to allow the clinician to see the interpretation of the given result. The execution period and the quality of analytical result depend on dialogue between analyst and clinician before and after analysis.

[Temporary antibiotic-loaded cemented prosthesis for two-stage septic hip arthroplasty]. / Prothèse temporaire en ciment imprégné d'antibiotiques au cours des arthroplasties en deux temps sur hanche infectée.

PURPOSE OF THE STUDY: During the excision period of a two-stage revision arthroplasty, the hip has a low function and an unacceptable leg length discrepancy. The goal of this study was to expose technical details in order to perform a simple articulated cement spacer which could be implanted during this period to improve hip function, to authorize partial weight bearing and to avoid leg length discrepancy. MATERIAL: This method was applied in three two-stage procedures justified because of particular immunodeficiency conditions: a 43 years old man who had bone marrow allograft and immunosuppressive therapy because of leukemia suffering of subacute septic hip arthritis; a 58 years old man suffering of diabetes and active C-hepatitis who had a septic loosening of a total hip arthroplasty (THA); a 76 years old woman suffering of diabetes who had a third septic loosening of THA. METHOD: The prosthesis was made of antibiotic-impregnated cement according to organisms antibiotic resistance. The prosthetic junction between head and diaphysis was reinforced with a tibial plate. Prosthetic shape was identical to the one of femoral broaches inserted in the femur after prosthetic and cement removal. The broach size was chosen when mechanical stability in the femur was obtained, and avoided leg length discrepancy after trials with cups. The tibial plate was bent in order to reinforce the junction with regard to the shape of the determined broach. Two doses of antibiotic-impregnated cement were mixed and molded with hands, then the plate was incorporated at the appropriate location, finally the broach was applied on this composite and cement in excess was removed before polymerisation. For prosthetic head, two options were available: to mold the cephalic zone of the cement at the patient acetabulum diameter with a soft aluminium cup previously molded in the acetabulum; to mold the cement cephalic zone with a trial cup in order to obtain a 22 or 28 ball. For this last option, a third dose of antibiotic-impregnated cement was prepared and placed in the acetabulum, a trial femoral head was applied in it to mold the location for the 22 or 28 prosthetic head. Before insertion, a collar was applied on the stem to prevent migration. Active mobilization was encouraged, and partial weight-bearing authorized. RESULTS: The mean range of hip flexion during period was 60 degrees. The patients were discharged approximatively 12 days after the first stage. Two patients had effective painless partial weight-bearing. The second stage was performed six weeks later on the average. The second procedure was easier than the second stage of a conventional two-stage procedure because of: easy and low hemorrhagic dissection authorized by the prosthesis; low difficulties with soft tissue tension as the prosthesis prevents leg length discrepancy; preservation of the articular space which prevents soft tissue sacrifice during the second stage. CONCLUSION: This simple technique is effective to prevent complications related to the excision period of a two-stage hip revision arthroplasty. Likewise, the economical aspect (short delay of hospitalisation, quick functional recovery) should be considered when compared with the excision period of a conventional two-stage procedure.