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International audience; Candida hispaniensis is an oleaginous yeast with a great potential for production ofsingle cell oil according to its naturally high lipid accumulation capacity. Its unusualsmall genome size trait is also attractive for fundamental research on genome evolu-tion. Our physiological study suggests a great potential for lipid production, reaching224 mg/g of cell dry weight in glucose minimum medium. C. hispaniensis is also ableto secrete up to 34.6 mg/L of riboflavin promising further riboflavin productionimprovements by cultivation optimization and genetic engineering. However, whileits genome sequence has been released very recently, no genetic tools have beendescribed up to now for this yeast limiting its use for fundamental research and forexploitation in an industrial biotechnology. We report here the first genetic modifica-tion of C. hispaniensis by introducing a heterologous invertase allowing the growth onsucrose using a biolistic transformation approach using a dedicated vector.The first genetic tool and transformation method developed here appear as a proofof concept, and while it would benefit from further optimization, heterogeneousexpression of invertase allows for metabolism of an additional sugar and shows het-erologous enzyme production capacity.

Even though production and open use of polychlorinated biphenyls (PCBs) have been phased out in Western industrialised countries since the 1980s, PCBs were still present in waste collected from different waste handling facilities in Norway in 2013. Sums of seven indicator-PCBs (I-PCB7:PCB-28, -52, -101, -118, -138, -153 and -180) were highest in plastic waste (3700 +/- 1800 mu g/kg, n=15), waste electrical and electronic equipment (WEEE) (1300 +/- 400 mu g/kg, n=12) and fine vehicle fluff (1800 = 1400 mu g/kg, n=4) and lowest in glass waste, combustibles, bottom ash and fly ash (0.3 to 65 mu g/kg). Concentrations in leachate water varied from 1.7 to 2900 ng/L, with higher concentrations found at vehicle and WEEE handling facilities. Particles in leachate water exhibited similar PCB sorption properties as solid waste collected on site, with waste-water partitioning coefficients ranging from 10(5)to 10(7) .I-PCB7 in air samples collected at the sites were mostly in the gas phase (100-24000 pg/m(3)), compared to those associated with particles (9-1900 pg/m(3)). In contrast, brominated flame retardants (BFRs) in the same samples were predominantly found associated with particles (e.g. sum of 10 brominated diethyl ethers, Sigma BDE10, associated with particles 77-194,000 pg/m(3)) compared to the gas phase (Sigma DE10 6-473 pg/m(3)). Measured gas-phase I-PCB7 concentrations are less than predicted, assuming waste-air partitioning in equilibrium with predominant waste on site. However, the gas-particle partitioning behavior of PCBs and BFRs could be predicted using an established partitioning model for ambient aerosols. PCB emissions from Norwegian waste handling facilities occurred primarily in the form of atmospheric vapor or leachate particles.

Glenoid lateralization has been shown to be the most imortant factor in maximizing passive range of motion and shoulder stability while preventing scapular impingement and notching. We aimed to evaluate mid- to long-term functional and radiologic outcomes after bony increased-offset-reverse shoulder arthroplasty (BIO-RSA) using a Grammont-style (medialized) humeral implant.The study included 143 consecutive shoulders (140 patients; mean age, 72 years) treated with a BIO-RSA for rotator cuff deficiency. A cylinder of autologous cancellous bone graft, harvested from the humeral head, was placed between the reamed glenoid surface and baseplate; fixation was achieved using a long central peg (25 mm) and 4 screws. A large baseplate (29 mm) with a small baseplate sphere (36 mm) was used in 77% of cases, and a Grammont-style (medialized) humeral implant with 155° of inclination was used in all cases. All patients underwent clinical and radiographic assessment at a minimum of 5 years after surgery; in addition, 86 cases (60%) underwent computed tomography scan assessment.At a mean follow-up of 75 months (range, 60-126 months), the survivorship of the BIO-RSA using revision as an endpoint was 96%. No cases of dislocation or humeral loosening were observed. Overall, 118 patients (83%) were either very satisfied (61%) or satisfied (22%). The adjusted Constant score improved from 40% ± 18% to 93% ± 23%, and the Subjective Shoulder Value improved from 31% ± 15% to 77% ± 18% (P.001). The humeral bone graft incorporated completely in 96% of cases (137 of 143). Severe inferior scapular notching (grade 3 or 4) occurred in 18% (24 of 136). The risk of postoperative notching correlated to a lower body mass index (P.05), superior glenosphere inclination (P = .02), and high or flush glenosphere positioning (P = .035).BIO-RSA is a safe and effective technique to lateralize the glenoid, providing consistent bone graft healing, excellent functional outcomes, a low revision rate, and a high rate of patient satisfaction. Thin patients (with a low body mass index) and glenosphere malposition (with persistent superior inclination and/or insufficient lowering of the baseplate and sphere) are associated with higher risk of scapular notching.Our data confirm the importance of implanting the baseplate with a neutral inclination (reverse shoulder arthroplasty angle5°) and with sufficient glenosphere inferior overhang (5 mm) in preventing scapular notching. The use of a less medialized humeral implant (135° or 145° inclined) and smaller (25-mm) baseplate (when using a small, 36-mm sphere) should allow reduction in the incidence of scapular notching.

Current initiatives for waste-handling in a circular economy favor prevention and recycling over incineration or landfilling. However, the impact of such a transition on environmental emissions of contaminants like bisphenol A (BPA) during waste-handling is not fully understood. To address this, a material flow analysis (MFA) was constructed for selected waste categories in Norway, for which the amount recycled is expected to increase in the future; glass, vehicle, electronic, plastic and combustible waste. Combined, 92 tons/y of BPA are disposed of via these waste categories in Norway, with 98.5% associated with plastic and electronic waste. During the model year 2011, the MFA showed that BPA in these waste categories was destroyed through incineration (60%), exported for recycling into new products (35%), stored in landfills (4%) or released into the environment (1%). Landfilling led to the greatest environmental emissions (up to 13% of landfilled BPA), and incinerating the smallest (0.001% of incinerated BPA). From modelling different waste management scenarios, the most effective way to reduce BPA emissions are to incinerate BPA-containing waste and avoid landfilling it. A comparison of environmental and human BPA concentrations with CoZMoMAN exposure model estimations suggested that waste emissions are an insignificant regional source. Nevertheless, from monitoring studies, landfill emissions can be a substantial local source of BPA. Regarding the transition to a circular economy, it is clear that disposing of less BPA-containing waste and less landfilling would lead to lower environmental emissions, but several uncertainties remain regarding emissions of BPA during recycling, particularly for paper and plastics. Future research should focus on the fate of BPA, as well as BPA alternatives, in emerging reuse and recycling processes, as part of the transition to a circular economy. © 2017. This is the authors’ accepted and refereed manuscript to the article. Locked until 13.1.2019 due to copyright restrictions. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/

OBJECT Unilateral subthalamotomy is a surgical procedure that may be used to alleviate disabling levodopa-induced dyskinesias (LIDs) in patients with Parkinson disease (PD). However, the mechanisms involved in LID remain largely unknown. The subthalamic nucleus (STN) is the sole glutamatergic nucleus within the basal ganglia, and its lesion may produce changes in glutamate receptors in various areas of the basal ganglia. The authors aimed to investigate the biochemical changes in glutamate receptors in striatal and pallidal regions of the basal ganglia after lesion of the STN in parkinsonian macaque monkeys. METHODS The authors treated 12 female ovariectomized monkeys with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to induce PD-like symptoms, treated 8 of these animals with 3,4-dihydroxy-l-phenylalanine (L-DOPA; levodopa) to induce LID, and performed unilateral subthalamotomy in 4 of these 8 monkeys. Four additional monkeys were treated with saline only and were used as controls. The MPTP monkeys had previously been shown to respond behaviorally to lower doses of levodopa after the STN lesion. Autoradiography of slices from postmortem brain tissues was used to visualize changes in the specific binding of striatal and pallidal ionotropic glutamate receptors (that is, of the α-amino-3-hydroxy 5-methyl-4-isoxazole propionate [AMPA] and N-methyl-d-aspartate [NMDA] NR1/NR2B subunit receptors) and of metabotropic glutamate (mGlu) receptors (that is, mGlu2/3 and mGlu5 receptors). The specific binding and distribution of glutamate receptors in the basal ganglia of the levodopa-treated, STN-lesioned MPTP monkeys were compared with those in the saline-treated control monkeys and in the saline-treated and levodopa-treated MPTP monkeys. RESULTS The autoradiographic results indicated that none of the pharmacological and surgical treatments produced changes in the specific binding of AMPA receptors in the basal ganglia. Levodopa treatment increased the specific binding of NMDA receptors in the basal ganglia. Subthalamotomy reversed these increases in the striatum, but in the globus pallidus (GP), the subthalamotomy reversed these increases only contralaterally. Levodopa treatment reversed MPTP-induced increases in mGlu2/3 receptors only in the GP. mGlu2/3 receptor–specific binding in the striatum and GP decreased bilaterally in the levodopa-treated, STN-lesioned MPTP monkeys compared with the other 3 groups. Compared with mGlu5 receptor–specific binding in the control monkeys, that of the levodopa-treated MPTP monkeys increased in the dorsal putamen and remained unchanged in the caudate nucleus and in the GP. CONCLUSIONS These results implicate glutamate receptors in the previously observed benefits of unilateral subthalamotomy to improve motor control.

International audience; The "pharmaceutical" polar organic integrative sampler (POCIS) is a passive sampler composed of an outer polyethersulfone (PES) membrane and an inner receiving Hydrophilic-Lipophilic Balance (HLB) phase. Target micropollutants can accumulate in the POCIS HLB phase following different uptake patterns. Two of the most common ones are a first-order kinetic uptake (Chemical Reaction Kinetic 1, CRK1 model), and a first-order kinetic uptake with an inflexion point (CRK2 model). From a previous study, we identified 30 and 13 micropollutants following CRK1 and CRK2 accumulation model in the POCIS HLB phase, respectively. We hypothesized that uptake in the outer PES membrane of POCIS may influence the uptake pathway. Thus, novel measurements of uptake in PES membrane were performed for these micropollutants to characterise kinetic accumulation in the membrane with and without the HLB phase. We determined, for the first time, the membrane-water distribution coefficient for 31 micropolluants. Moreover, the lag times for molecules to breakthrough the POCIS membrane increased with increasing hydrophobicity, defined by the octanol-water dissociation constant Dow. However, Dow alone was insufficient to predict whether uptake followed a CRK1 or CRK2 model in the POCIS HLB phase. Thus, we performed a factorial discriminant analysis considering several molecular physico-chemical properties, and the model of accumulation for the studied micropollutants can be predicted with > 90% confidence. The most influent properties to predict the accumulation model were the log Dow and the polar surface area of the molecule (> 70% confidence with just these two properties). Molecules exhibiting a CRK1 uptake model for the POCIS HLB phase tended to have log Dow > 2.5 and polar surface area < 50 Ǻ2.

Background The aims of this study were to determine the survival of anatomic total shoulder arthroplasty with uncemented metal-backed (MB) glenoid components with a polyethylene (PE) insert in primary osteoarthritis, to assess the reasons for revision surgery, and to identify patients and diagnostic factors that influence failure rates. Methods Between 1994 and 1999, 165 patients (mean age, 68 years) with primary osteoarthritis were treated with anatomic total shoulder arthroplasty using an uncemented MB/PE glenoid component. Outcomes were assessed both clinically and radiologically with a minimum of 2 years of follow-up. A prosthetic survival curve was constructed with the end point defined as either partial or complete revision, using 100% confidence intervals. Results Survival rate free of revision was 46% (100% confidence interval, 32%-54%) at 12 years. At a mean follow-up of 8.5 years (range, 2-16 years), revision was required in 61 patients (37%); 80% of shoulders undergoing revision (49 of 61) had evidence of PE wear. Glenoid loosening (because of osteolysis secondary to wear debris), soft tissue deficiency, and prosthetic instability were the most common modes of failure. Younger patients and biconcave glenoids (with posterior humeral subluxation) have a negative effect on implant survival. Proximal humerus osteolysis was significantly more frequent in shoulders with PE wear. Exchange of the PE insert (with conservation of the MB tray) was possible in only 3% of the revised shoulders. Conclusion Uncemented MB glenoid resurfacing is not a viable long-term therapeutic option because of accelerated PE wear leading to early revision surgery. Conservation of the MB tray with reinsertion of a new PE insert is rarely possible because of glenoid bone loss, implant loosening, soft tissue deficiency, and prosthetic instability. Younger patients and biconcave glenoids have a negative effect on implant survival.

The plastic additive bisphenol A (BPA) is commonly found in landfill leachate at levels exceeding acute toxicity benchmarks. To gain insight into the mechanisms controlling BPA emissions from waste and waste-handling facilities, a comprehensive field and laboratory campaign was conducted to quantify BPA in solid waste materials (glass, combustibles, vehicle fluff, waste electric and electronic equipment (WEEE), plastics, fly ash, bottom ash, and digestate), leachate water, and atmospheric dust from Norwegian sorting, incineration, and landfill facilities. Solid waste concentrations varied from below 0.002 mg/kg (fly ash) to 188 ± 125 mg/kg (plastics). A novel passive sampling method was developed to, for the first time, establish a set of waste-water partition coefficients, KD,waste, for BPA, and to quantify differences between total and freely dissolved concentrations in waste-facility leachate. Log-normalized KD,waste (L/kg) values were similar for all solid waste materials (from 2.4 to 3.1), excluding glass and metals, indicating BPA is readily leachable. Leachate concentrations were similar for landfills and WEEE/vehicle sorting facilities (from 0.7 to 200 μg/L) and dominated by the freely dissolved fraction, not bound to (plastic) colloids (agreeing with measured KD,waste values). Dust concentrations ranged from 2.3 to 50.7 mg/kgdust. Incineration appears to be an effective way to reduce BPA concentrations in solid waste, dust, and leachate.

Subthalamotomy allows a reduction of doses of l-DOPA in dyskinetic patients while its antiparkinsonian benefits are preserved. However, the mechanisms of the potentiation of this response to medication remain to be elucidated. Hence, dopamine D1 and D2 receptors as well as the dopamine transporter were investigated using receptor binding autoradiography. D1 and D2 receptors as well as preproenkephalin and preprodynorphin mRNA levels were measured by in situ hybridization. Four dyskinetic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) parkinsonian monkeys that underwent unilateral subthalamotomy were compared to four controls, four saline-treated and four l-DOPA-treated MPTP monkeys. Dopamine, its metabolites and its transporter were extensively and similarly decreased in all parkinsonian monkeys. D1 receptor specific binding was decreased in the striatum of all MPTP monkeys. The l-DOPA-induced decrease in D1 receptor specific binding was reversed in the striatum ipsilateral to subthalamotomy. D1 receptor mRNA levels followed a similar pattern. D2 receptor specific binding and mRNA levels remained unchanged in all groups. Striatal preproenkephalin mRNA levels were overall increased in MPTP monkeys; the STN-lesioned parkinsonian group had significantly lower values than the saline-treated and l-DOPA-treated parkinsonian monkeys in the dorsolateral putamen. Striatal preprodynorphin mRNA levels remained unchanged in MPTP monkeys compared to controls whereas it increased in all monkeys treated with l-DOPA compared to controls; subthalamotomy induced a decrease in the dorsolateral putamen ipsilateral to surgery. The improved motor response to l-DOPA after subthalamotomy in the parkinsonian monkeys investigated may be associated with an increased synthesis and expression of D1 receptors ipsilateral to STN lesion of the direct pathway.

The dopamine transporter (DAT) is abundantly expressed in the striatum where it removes extracellular dopamine into the cytosol of presynaptic nerve terminals. It is the target of drugs of abuse and antidepressants. There is a loss of the DAT in Parkinson's disease affecting release of levodopa implicated in levodopa-induced dyskinesias. This study investigated the effect of cholesterol on DAT, serotonin transporter (SERT) and vesicular monoamine transporter 2 (VMAT2) in monkey and rat brains in vitro. DAT protein levels measured by Western blot remained unchanged with in vitro methyl-β-cyclodextrin (MCD) incubations to remove membrane cholesterol or with incubations to increase membrane cholesterol content. By contrast, striatal DAT specific binding labelled with [125I]RTI-121 or with [125I]RTI-55 decreased with increasing concentrations of MCD and increased with cholesterol loading. Moreover, [125I]RTI-121 specific binding of striatal membranes depleted of cholesterol with MCD was restored to initial DAT content with addition of cholesterol showing its rapid and reversible effect. By contrast, striatal VMAT2 and SERT specific binding showed no or limited changes by cholesterol manipulations. Similar results were obtained for monkey caudate nucleus, putamen and nucleus accumbens. Membrane microviscosity was assessed by fluorescence polarization spectroscopy, using the probe 1,6-diphenyl-1,3,5-hexatriene. DAT changes positively correlated with changes of membrane microviscosity in rat and monkey brain regions investigated and with membrane cholesterol contents. Similar findings were observed with desmosterol but to a lower extent than with cholesterol. These results show an important effect of cholesterol on the DAT associated with microviscosity changes that should be considered in drug therapies.

Flame retardants in commercial products eventually make their way into the waste stream. Herein the presence of flame retardants in Norwegian landfills, incineration facilities and recycling sorting/defragmenting facilities is investigated. These facilities handled waste electrical and electronic equipment (WEEE), vehicles, digestate, glass, combustibles, bottom ash and fly ash. The flame retardants considered included polybrominated diphenyl ethers (∑BDE-10) as well as dechlorane plus, polybrominated biphenyls, hexabromobenzene, pentabromotoluene and pentabromoethylbenzene (collectively referred to as ∑FR-7). Plastic, WEEE and vehicles contained the largest amount of flame retardants (∑BDE-10: 45,000-210,000μg/kg; ∑FR-7: 300-13,000μg/kg). It was hypothesized leachate and air concentrations from facilities that sort/defragment WEEE and vehicles would be the highest. This was supported for total air phase concentrations (∑BDE-10: 9000-195,000pg/m

Polar organic chemical integrative samplers (POCIS) have previously been used to monitor alkylphenol (AP) contamination in water and produced water. However, only the sorbent receiving phase of the POCIS (Oasis beads) is traditionally analyzed, thus limiting the use of POCIS for monitoring a range of APs with varying hydrophobicity. Here a "pharmaceutical" POCIS was calibrated in the laboratory using a static renewal setup for APs (from 2-ethylphenol to 4-n-nonylphenol) with varying hydrophobicity (log Kow between 2.47 and 5.76). The POCIS sampler was calibrated over its 28 day integrative regime and sampling rates (Rs) were determined. Uptake was shown to be a function of AP hydrophobicity where compounds with log Kow < 4 were preferentially accumulated in Oasis beads, and compounds with log Kow > 5 were preferentially accumulated in the PES membranes. A lag phase (over a 24 h period) before uptake in to the PES membranes occurred was evident. This work demonstrates that the analysis of both POCIS phases is vital in order to correctly determine environmentally relevant concentrations owing to the fact that for APs with log Kow ≤ 4 uptake, to the PES membranes and the Oasis beads, involves different processes compared to APs with log Kow ≥ 4. The extraction of both the POCIS matrices is thus recommended in order to assess the concentration of hydrophobic APs (log Kow ≥ 4), as well as hydrophilic APs, most effectively.

Sugar assimilation has been intensively studied in the model yeast S. cerevisiae, and for two decades, it has been clear that the homologous HXT genes, which encode a set of hexose transporters, play a central role in this process. However, in the yeast Yarrowia lipolytica, which is well-known for its biotechnological applications, sugar assimilation is only poorly understood, even though this yeast exhibits peculiar intra-strain differences in fructose uptake: some strains (e.g., W29) are known to be slow-growing in fructose while others (e.g., H222) grow rapidly under the same conditions. Here, we retrieved 24 proteins of the Sugar Porter family from these two strains, and determined that at least six of these proteins can function as hexose transporters in the heterologous host Saccharomyces cerevisiae EBY.VW4000. Transcriptional studies and deletion analysis in Y. lipolytica indicated that two genes, YHT1 and YHT4, are probably the main players in both strains, with a similar role in the uptake of glucose, fructose, and mannose at various concentrations. The other four genes appear to constitute a set of 'reservoir' hexose transporters with an as-yet unclear physiological role. Furthermore, through examining Sugar Porters of the entire Yarrowia clade, we show that they constitute a dynamic family, within which hexose transport genes have been duplicated and lost several times. Our phylogenetic analyses support the existence of at least three distinct evolutionary groups of transporters which allow yeasts to grow on hexoses. In addition to the well-known and widespread Hxt-type transporters (which are not essential in Y. lipolytica), we highlight a second group of transporters, represented by Yht1, which are phylogenetically related to sensors that play a regulatory role in S. cerevisiae, and a third group, represented by Yht4, previously thought to contain only high-affinity glucose transporters related to Hgt1of Kluyveromyces lactis.

Glutamate overactivity is well documented in Parkinson’s disease (PD) and dyskinesias induced by L-3,4-dihydroxyphenylalanine (L-DOPA), the gold-standard treatment for this disease. The contribution of metabotropic glutamate receptors type 5 (mGlu5 receptors) in PD and L-DOPA-induced dyskinesias (LID) was the topic of investigations in human PD patients and in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) monkeys.

The treatment of motor symptoms of Parkinson disease (PD) with the dopamine (DA) precursor, l -3,4-dihydroxyphenylalanine ( l -DOPA) introduced 50 years ago still remains a very effective medication. However, involuntary movements termed l -DOPA-induced dyskinesias (LID) appear in the vast majority of PD patients after chronic treatment and may become disabling. Once they appeared, the first dose after a several-weeks drug holiday will trigger them again, showing that l -DOPA has permanently or persistently modified the brain response to DA. LID are very difficult to manage and no drug is yet approved for dyskinesias, aside from a modest benefit with amantadine. New drugs are needed for PD to alleviate parkinsonian symptoms without inducing dyskinesias. Hence, animal models have been developed to seek the mechanisms involved in LID and new drug targets. The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was discovered as a contamination of a derivative of heroin taken by drug users and produced similar motor symptoms as idiopathic PD. Since then, MPTP is used extensively to model PD and LID in non-human primates and mice in addition to the classical PD model in rats with a 6-hydroxydopamine (6-OHDA) lesion. This article reviews rodent and non-human primate models of PD that reproduce motor complications induced by DA replacement therapy. Moreover, key biochemical changes in the brain of post-mortem PD patients with LID will be compared to those observed in animal models. Finally, the translational usefulness of drugs found to treat LID in animal models will be compared to their clinical activities.

Glenoid deficiency and erosion (excessive retroversion/inclination) must be corrected in reverse shoulder arthroplasty (RSA) to avoid prosthetic notching or instability and to maximize function, range of motion, and prosthesis longevity. This study reports the results of RSA with an angled, autologous glenoid graft harvested from the humerus (angled BIO-RSA).A trapezoidal bone graft, harvested from the humeral head and fixed with a long-post baseplate and screws, was used to compensate for residual glenoid bone loss/erosion. For simple to moderate (25°) glenoid defects, standardized instrumentation combined with some eccentric reaming (15°) was used to reconstruct the glenoid and obtain neutral implant alignment. For severe (25°) and complex (multiplanar) glenoid bone defects, patient-specific grafts and guides were used after 3-dimensional planning. Patients were reviewed with minimum 2 years of follow-up. Mean follow-up was 36 months (range, 24-81 months). Preoperative and postoperative measurements of inclination and version were performed in the plane of the scapula on computed tomography images.The study included 54 patients (41 women, 13 men; mean 73 years old). Fifteen patients had combined vertical and horizontal glenoid bone deficiency. Among E2/E3 glenoids, inclination improved from 37° (range, 14° to 84°) to 10.2° (range -28° to 36°, P .001). Among B2/C glenoids, retroversion improved from -21° (range, -49° to 0°) to -10.6° (-32° to 4°, P = .06). Complete radiographic incorporation of the graft occurred in 94% (51 of 54). Complications included infection in 1 and clinical aseptic baseplate loosening in 2. Mild notching occurred in 25% (13 of 51) of patients. Constant-Murley and Subjective Shoulder Value assessments increased from 31 to 68 and from 30% to 83%, respectively (P .001).Angled BIO-RSA predictably corrects glenoid deficiency, including severe (25°) multiplanar deformity. Graft incorporation is predictable. Advantages of using an autograftharvested in situ include bone stock augmentation, lateralization, low donor-site morbidity, low relative cost, and flexibility needed to simultaneously correct posterior and superior glenoid defects.

Metabotropic glutamate 5 (mGlu5) receptor antagonists reduce L-3,4-dihydroxyphenylalanine (L-DOPA)-induced dyskinesias (LID) in Parkinson's disease (PD). The aim of this study was to investigate the long-term effect of the prototypal mGlu5 receptor antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP) on glutamate receptors known to be involved in the development of LID in the de novo chronic treatment of monkeys lesioned with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). MPTP monkeys were treated for one month with L-DOPA and developed dyskinesias while those treated with L-DOPA and MPEP (10 mg/kg) developed significantly less. Normal control and saline-treated MPTP monkeys were also included. All MPTP monkeys were extensively and similarly denervated. The basal ganglia [(3)H]ABP688 specific binding (mGlu5 receptors) was elevated in L-DOPA-treated MPTP monkeys compared to controls but not in those treated with L-DOPA and MPEP; dyskinesia scores of these monkeys correlated positively with their [(3)H]ABP688 specific binding. Striatal density (B(max)) of [(3)H]ABP688 specific binding increased in L-DOPA-treated MPTP monkeys compared to other groups and affinity (Kd) remained unchanged. Striatal mGlu5 receptor mRNA remained unchanged following treatments. Elevated basal ganglia specific binding of [(3)H]Ro 25-6981 (NMDA NR1/NR2B receptors), [(3)H]Ro 48-8587 (AMPA receptors) but not [(3)H]CGP-39653 (NMDA NR1/NR2A receptors) was observed only in L-DOPA-treated MPTP monkeys; dyskinesias scores correlated with binding. By contrast, basal ganglia [(3)H]LY341495 specific binding (mGlu2/3 receptors) decreased in L-DOPA-treated MPTP monkeys compared to controls, saline and L-DOPA + MPEP treated MPTP monkeys; dyskinesias scores correlated negatively with this binding. Hence, chronic MPEP treatment reduces the development of LID and is associated with a normalization of glutamate neurotransmission.

l -3,4-Dihydroxyphenylalanine ( l -DOPA), the gold standard therapy for Parkinson disease (PD), is associated with motor fluctuations and dyskinesias. This study sought to prevent the development of l -DOPA-induced dyskinesias (LID) with the metabotropic glutamate receptor type 5 (mGlu5 receptor) antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP) in the de novo treatment of monkeys lesioned with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) as a PD model. MPTP-lesioned monkeys were treated once daily for one month with either l -DOPA or l -DOPA + MPEP (10 mg/kg). MPEP (administered 15 min before l -DOPA) plasma concentrations were elevated during all the l -DOPA motor activation and did not accumulate during a month. The antiparkinsonian effect was maintained throughout the treatment period in MPTP-lesioned monkeys treated with l -DOPA + MPEP, while the duration of this effect decreased over time in MPTP-lesioned monkeys treated with l -DOPA alone, suggesting wearing-off. Over the month-long treatment, the mean dyskinesia score increased in l -DOPA-treated monkeys; interestingly, this increase was reduced by overall 72% in the l -DOPA + MPEP group. Mean dyskinesia scores of monkeys correlated inversely with plasma MPEP concentrations. Normal control and saline-treated MPTP-lesioned monkeys were also included for biochemical analyses. All MPTP-lesioned monkeys were extensively and similarly denervated. [3H]ABP688 specific binding to mGlu5 receptors increased in the putamen of l -DOPA-treated monkeys compared to control, saline or l -DOPA + MPEP-treated monkeys. Mean dyskinesia scores of MPTP-lesioned monkeys correlated positively with [3H]ABP688 specific binding in the putamen. This study showed a beneficial chronic antidyskinetic effect of MPEP in de novo l -DOPA-treated MPTP-lesioned monkeys, supporting the therapeutic use of mGlu5 receptor antagonists in PD to prevent LID. This article is part of a Special Issue entitled ‘Metabotropic Glutamate Receptors’.

Object Subthalamotomy is a stereotactic surgery performed in patients with disabling dyskinesias due to Parkinson disease. The authors set out to model this human condition in MPTP monkeys and determine if subthalamotomy allowed a reduction of levodopa for similar benefit. Methods The authors performed unilateral subthalamotomy in 4 parkinsonian dyskinetic monkeys by stereotactic injection of ibotenic acid. An optimal dose, defined as the highest dose of levodopa improving parkinsonian motor symptoms while inducing low or no dyskinesias, was established in these animals. Each monkey was scored for the antiparkinsonian and dyskinetic effects of the optimal dose of levodopa, as well as suboptimal and dyskinesia-inducing doses (60% and 140% of the optimal dose, respectively), and these scores were compared with those obtained at baseline before and after subthalamotomy. Bradykinesia was assessed by a prehension task. Results Unilateral subthalamotomy had a positive effect on the antiparkinsonian response for all doses of levodopa as well as the baseline. There were no differences in the antiparkinsonian response between the suboptimal dose postsurgery and the optimal dose presurgery. Dyskinesias were increased at the suboptimal and the optimal doses. After surgery, the duration of response to levodopa increased between 20% and 25% in the suboptimal dose, whereas it remained unchanged with higher doses. Bradykinesia was significantly reduced after surgery only at the suboptimal dose. Conclusions Subthalamotomy potentiated the response to suboptimal doses of levodopa. Thus, levodopa can be reduced by 40% after surgery for similar beneficial antiparkinsonian response and less dyskinesia than with an optimal dose before surgery.

L-DOPA-induced dyskinesias (LID) appear in the majority of Parkinson's disease (PD) patients. Nicotinic acetylcholine (nACh) receptor-mediated signaling has been implicated in PD and LID and modulation of brain α7 nACh receptors might be a potential therapeutic target for PD. This study used [(125)I]α-Bungarotoxin autoradiography to investigate α7 nACh receptors in LID in post-mortem brains from PD patients (n=14) and control subjects (n=11), and from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkeys treated with saline (n=5), L-DOPA (n=4) or L-DOPA+2-methyl-6-(phenylethynyl)pyridine (MPEP) (n=5), and control monkeys (n=4). MPEP is the prototypal metabotropic glutamate 5 (mGlu5) receptor antagonist; it reduced the development of LID in these monkeys. [(125)I]α-Bungarotoxin specific binding to striatal and pallidal α7 nACh receptors were only increased in L-DOPA-treated dyskinetic MPTP monkeys as compared to controls, saline and L-DOPA+MPEP MPTP monkeys; dyskinesia scores correlated positively with this binding. The total group of Parkinsonian patients had higher [(125)I]α-Bungarotoxin specific binding compared to controls in the caudate nucleus but not in the putamen. PD patients without motor complications had higher [(125)I]α-Bungarotoxin specific binding compared to controls only in the caudate nucleus. PD patients with LID only had higher [(125)I]α-Bungarotoxin specific binding compared to controls in the caudate nucleus and compared to those without motor complications and controls in the putamen. PD patients with wearing-off only, had [(125)I]α-Bungarotoxin specific binding at control values in the caudate nucleus and lower in the putamen. Reduced motor complications were associated with normal striatal α7 nACh receptors, suggesting the potential of this receptor to manage motor complications in PD.

The number of fully sequenced genomes of yeasts is dramatically increasing but both structural and functional annotation quality are usually neglected, as most frequently based on automatic annotation transfer tools from reference genomes. RNA sequencing technologies offer the possibility to better characterize yeast transcriptomes and to correct or improve the prediction of mRNA, ncRNA, or miscellaneous RNA. We describe a computational approach to enhance structural annotation of yeast genomes based on RNA-Seq data exploitation. The proposed pipeline is primarily based on read mapping with TopHat2. Mapping outputs are then used for various applications such as: (1) validation of exon-exon junctions of predicted transcripts, (2) definition of new transcribed features, (3) prediction of 3' UTR, and (4) identification of extra features absent from the genome assembly. We strongly encourage curators to proceed to a manual validation and editing of the reference genome. Releasing genomes with high-quality annotation is an important issue, as they will be considered as references for further predictions.

The genus Arthrospira comprises filamentous cyanobacteria in which the trichomes form an open helix and contain gas vacuoles. The gas vesicle gene cluster of five Arthrospira strains was amplified by PCR and sequenced. The genes are organized in one operon, in the order gvpA1–gvpC1–gvpA2–gvpC2–gvpA3–gvpC3–gvpN. In Arthrospira sp. strain PCC 8005, the genes gvpJ, gvpK, gvpV and gvpW were also identified. Each of the three copies of gvpA encodes a protein of 71 amino acids. In the case of gvpC, there are two different length variants. Each of the two shorter genes, gvpC1 and gvpC2, encodes a putative protein of 151 amino acids, while the longer one, gvpC3, codes for a putative protein of 284 residues. The amino acid sequences of GvpC1 and GvpC2 are identical to the N-terminal part of GvpC3. In spite of the presence of stop codons downstream of gvpC1 and gvpC2, the deduced amino acid sequences in these regions are highly similar to the C-terminal part of GvpC3 (residues 160 to 229). The GvpC1, GvpC2 and GvpC...

POCIS (Polar Organic Chemical Integrative Sampler) is a relatively recent integrative sampler developed to trap hydrophilic organic micropollutants in aquatic environments. Nevertheless, at present, there is no review dealing specifically with this tool. The aim of this paper was to compile information from numerous references based on POCIS in order to discuss on the evaluation of time-weighted average concentrations (calibration methods, sampling rates, performance and reference compounds...) and to critically review the different in situ applications (screening or quantifying micropollutants, coupling with toxicity tests), application domains (molecules analyzed, sampling media) and analytical protocols for POCIS (processing, analysis, exposure duration).

International audience; The uptake kinetics of 27 emerging pollutants on the polar C18 Chemcatcher have been investigated. This investigation determined the sampling rates of 20 compounds, including 16 endocrine disruptors and 4 pharmaceuticals, which were used as overall pollution indicators. Calibrations were completed in a 50-L flow-through microcosm with continuous renewal of tap water spiked with approximately 3 μg/L of each pollutant and with sampling times at 1, 3, 6 and 12 h and 1, 3, 7, 14, 21 and 28 days. Exponential regressions for the accumulation kinetics were plotted to confirm the maximum linear uptake times for each molecule using the half time of equilibrium (t1/2) criteria. Of the compounds tested, 17 were accumulated linearly for up to 14 or 21 days with an R2 above 0.98 for linear correlations. The evaluation of the release kinetics of a C18 Chemcatcher spiked with 20 deuterated compounds identified 3 potential performance reference compounds (PRCs) with exponential desorption rates showing relatively good isotropic exchange. Highlights Uptake kinetics of 27 emerging pollutants were investigated on the polar C18 Chemcatcher. Sampling rates of 20 endocrine disruptors were determined. 17 compounds accumulated linearly up to 14 or 21 days. 3 potential PRCs on C18 Chemcatcher were identified.

P2Y14 is a member of the pyrimidinergic GPCR family. UDP-Glc has been previously shown to activate human P2Y14, whereas UDP was unable to activate the receptor. In this study, the authors used conventional and nonconventional methods to further characterize P2Y14 and its ligands. Conventional calcium mobilization and nonconventional cellular impedance functional assays revealed that UMP and UDP selectively activated HEK cells coexpressing P2Y14 and Gα(qi5). In the impedance assays, the presence of exogenous Gα(qi5) resulted in agonist-induced Gq signaling, whereas in the absence of exogenous Gα(qi5), the signal was indicative of Gi. The authors established the first P2Y14 membrane filtration binding assay using a novel optimized expression vector and [(3)H]UDP as radioligand. UDP-Glc, UMP, and UDP dose dependently inhibited [(3)H]UDP binding in the binding assay, and saturation analysis revealed that UDP bound P2Y14 with a K(D) = 10 nM and a B(max) = 110 pmol/mg. The authors screened a phosphonate library and identified compound A, which inhibited UDP-Glc-mediated calcium signaling in the fluorometric imaging plate reader assay (IC(50) = 2.3 µM) and competed for [(3)H]UDP binding in the novel binding assay with a K(i) = 1280 nM.

In recent years, metal oxides such as titania have been commercially available as chromatographic beds that can potentially be used to achieve novel separations of polar compounds. For example beta blockers, which are more often encountered in environmental sciences, have a wide range of polarity, and their basic character leads to difficult sample treatment and separation on conventional silica-based sorbents. The contribution of titania to the selective analysis of nine beta blockers was evaluated in terms of retention mechanisms observed in hydrophilic interaction LC using acetonitrile/water mobile phases with various additives. The mobile phase additives enabled to control the beta blocker charge as well as the titania surface charge. Depending on their respective ionic state, various retention mechanisms were identified at low water contents (

A weak, UDP-competitive antagonist of the pyrimidinergic receptor P2RY(14) with a naphthoic acid core was identified through high-throughput screening. Optimization provided compounds with improved potency but poor pharmacokinetics. Acylglucuronidation was determined to be the major route of metabolism. Increasing the electron-withdrawing nature of the substituents markedly reduced glucuronidation and improved the pharmacokinetic profile. Additional optimization led to the identification of compound 38 which is an 8 nM UDP-competitive antagonist of P2Y(14) with a good pharmacokinetic profile.

A weak antagonist of the pyrimidinergic receptor P2Y(14) containing a dihydropyridopyrimidine core was identified through high-throughput screening. Subsequent optimization led to potent, non-UTP competitive antagonists and represent the first reported non-nucleotide antagonists of this receptor. Compound 18q was identified as a 10 nM P2Y(14) antagonist with good oral bioavailability and provided sufficient exposure in mice to be used as a tool for future in vivo studies.

Thanks to their photosynthetic and nutritive properties, cyanobacteria of the Arthrospira genus are of interest as food supplements, as efficient oxygen producing life support system organisms for manned space flight, and for the production of biofuels. Despite these potential valuable applications, full genome sequences and genetic information in general on Arthrospira remain scarce. This is mainly due to the difficulty to extract sufficient high molecular weight nucleic acids from these filamentous cyanobacteria. In this article, an efficient and reproducible DNA extraction procedure for cyanobacteria of the genus Arthrospira was developed. The method is based on the combination of a soft mechanical lysis with enzymatic disruption of the cell wall. The comparison with other extraction protocols clearly indicates that this optimised method allows the recovery of a larger amount of DNA. Furthermore, the extracted DNA presents a high molecular weight, a reduced degradation and an excellent overall quality. It can be directly used for molecular biology purposes such as PCR, and clone library construction.

OBJECTIVESignificant new data suggest that metabolic disorders such as diabetes, obesity, and atherosclerosis all posses an important inflammatory component. Infiltrating macrophages contribute to both tissue-specific and systemic inflammation, which promotes insulin resistance. The complement cascade is involved in the inflammatory cascade initiated by the innate and adaptive immune response. A mouse genomic F2 cross biology was performed and identified several causal genes linked to type 2 diabetes, including the complement pathway.RESEARCH DESIGN AND METHODSWe therefore sought to investigate the effect of a C3a receptor (C3aR) deletion on insulin resistance, obesity, and macrophage function utilizing both the normal-diet (ND) and a diet-induced obesity mouse model.RESULTSWe demonstrate that high C3aR expression is found in white adipose tissue and increases upon high-fat diet (HFD) feeding. Both adipocytes and macrophages within the white adipose tissue express significant amounts of C3aR. C3aR−/− mice on HFD are transiently resistant to diet-induced obesity during an 8-week period. Metabolic profiling suggests that they are also protected from HFD-induced insulin resistance and liver steatosis. C3aR−/− mice had improved insulin sensitivity on both ND and HFD as seen by an insulin tolerance test and an oral glucose tolerance test. Adipose tissue analysis revealed a striking decrease in macrophage infiltration with a concomitant reduction in both tissue and plasma proinflammatory cytokine production. Furthermore, C3aR−/− macrophages polarized to the M1 phenotype showed a considerable decrease in proinflammatory mediators.CONCLUSIONSOverall, our results suggest that the C3aR in macrophages, and potentially adipocytes, plays an important role in adipose tissue homeostasis and insulin resistance.

The environmental behaviour of antimony (Sb) is gathering attention due to its increasingly extensive use in various products, particularly in plastics. Because of this it may be expected that plastic waste is an emission source for Sb in the environment. This study presents a comprehensive field investigation of Sb concentrations in diverse types of waste from waste handling facilities in Norway. The wastes included waste electrical and electronic equipment (WEEE), glass, vehicle fluff, combustibles, bottom ash, fly ash and digested sludge. The highest solid Sb concentrations were found in WEEE and vehicle plastic (from 1238 to 1715 mg kg(-1)) and vehicle fluff (from 34 to 4565 mg kg(-1)). The type of acid used to digest the diverse solid waste materials was also tested. It was found that HNO3:HCl extraction gave substantially lower, non-quantitative yields compared to HNO3:HF. The highest water-leachable concentration for wastes when mixed with water at a 1 : 10 ratio were observed for plastic (from 0.6 to 2.0 mg kg(-1)) and bottom ash (from 0.4 to 0.8 mg kg(-1)). For all of the considered waste fractions, Sb(v) was the dominant species in the leachates, even though Sb(iii) as Sb2O3 is mainly used in plastics and other products, indicating rapid oxidation in water. This study also presents for the first time a comparison of Sb concentrations in leachate at waste handling facilities using both active grab samples and DGT passive samples. Grab samples target the total suspended Sb, whereas DGT targets the sum of free- and other chemically labile species. The grab sample concentrations (from 0.5 to 50 μg L(-1)) were lower than the predicted no-effect concentration (PNEC) of 113 μg L(-1). The DGT concentrations were substantially lower (from 0.05 to 9.93 μg L(-1)) than the grab samples, indicating much of the Sb is present in a non-available colloidal form. In addition, air samples were taken from the chimney and areas within combustible waste incinerators, as well as from the vent of WEEE sorting facility. The WEEE vent had the highest Sb concentration (from100 to 2200 ng m(-3)), which were orders of magnitude higher than the air surrounding the combustible shredder (from 25 to 217 ng m(-3)), and the incinerator chimney (from30 to 100 ng m(-3)). From these results, it seems evident that Sb from waste is not an environmental concern in Norway, and that Sb is mostly readily recovered from plastic and bottom ash.

L-DOPA-induced dyskinesias (LID) are abnormal involuntary movements limiting the chronic use of L-DOPA, the main pharmacological treatment of Parkinson's disease. Serotonin receptors are implicated in the development of LID and modulation of basal ganglia 5-HT1B receptors is a potential therapeutic alternative in Parkinson's disease. In the present study, we used receptor-binding autoradiography of the 5-HT1B-selective radioligand [3H]GR125743 to investigate possible contributions of changes in ligand binding of this receptor in LID in post-mortem brain specimens from Parkinson's disease patients (n=14) and control subjects (n=11), and from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkeys treated with saline (n=5), L-DOPA (n=4) or L-DOPA+2-methyl-6-(phenylethynyl)pyridine (MPEP) (n=5), and control monkeys (n=4). MPEP is the prototypal metabotropic glutamate 5 (mGlu5) receptor antagonist and has been shown to reduce the development of LID in these monkeys in a chronic treatment of one month. [3H]GR125743 specific binding to striatal and pallidal 5-HT1B receptors respectively were only increased in L-DOPA-treated MPTP monkeys (dyskinetic monkeys) as compared to controls, saline and L-DOPA+MPEP MPTP monkeys; dyskinesias scores correlated positively with this binding. Parkinson's disease patients with motor complications (L-DOPA-induced dyskinesias and wearing-off) had higher [3H]GR125743 specific binding compared to those without motor complications and controls in the basal ganglia. Reduction of motor complications was associated with normal striatal 5-HT1B receptors, suggesting the potential of this receptor for the management of motor complications in Parkinson's disease.

MELiSSA is a bioregenerative life support system designed by the European Space Agency (ESA) for the complete recycling of gas, liquid and solid wastes during long distance space exploration. The system uses the combined activity of different living organisms: microbial cultures in bioreactors, a plant compartment and a human crew. In this minireview, the development of a short-cut ecological system for the biotransformation of organic waste is discussed from a microorganism's perspective. The artificial ecological model--still in full development--that is inspired by Earth's own geomicrobiological ecosystem serves as an ideal study object on microbial ecology and will become an indispensable travel companion in manned space exploration.

Previous results from our laboratory have shown that 17beta-oestradiol prevents 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) striatal dopamine depletion. 17beta-oestradiol, oestriol and oestrone are the naturally occurring oestogens in humans. Using various dopamine markers, the present study investigated whether oestrone and oestriol such as 17beta-oestradiol have neuroprotective activity in MPTP-treated mice. Male mice were treated with 17beta-oestradiol, oestriol or oestrone for 5 days before and after MPTP administration, and were compared with nonlesioned mice receiving the same treatment. Striatal concentrations of dopamine and its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were assayed by high-performance liquid chromatography. Dopamine transporter (DAT) and vesicular monoamine transporter (VMAT2) specific binding were measured by autoradiography. DAT, VMAT2 and tyrosine hydroxylase mRNA levels were measured by in situ hybridisation. MPTP induced a loss of DAT and VMAT2 specific binding in the striatum and substantia nigra, as well as a decrease of VMAT2 mRNA in the substantia nigra. 17beta-oestradiol treatment prevented the loss of these dopaminergic markers, as well as striatal concentrations of dopamine, DOPAC and HVA. Mice receiving oestriol and oestrone showed catecholamine concentrations comparable to MPTP mice. Oestriol treatment had no effect on dopaminergic markers in MPTP mice whereas oestrone prevented striatal DAT loss and the decrease of VMAT2 mRNA in the substantia nigra. In nonlesioned mice, 17beta-oestradiol, oestriol or oestrone had no effect on all the dopaminergic markers investigated. In conclusion, a weak or a lack of effect of oestriol and oestrone was observed compared to 17beta-oestradiol in MPTP mice and none of these steroids had an effect in nonlesioned mice. A DAT and VMAT2 specific binding decrease after MPTP in the striatum and substantia nigra, as well as a decrease of substantia nigra VMAT2 mRNA, was observed and could be prevented by oestradiol.

Cet article porte sur la modélisation des culasses aluminium de moteurs diesel HDI en vue de prédire leur tenue en fatigue polycyclique. Celles-ci, avant d'être frettées, serrées sur le bloc moteur et utilisées en service, subissent un traitement thermique (trempe et revenu) afin d'améliorer leurs propriétés mécaniques. Lors de ce procédé d'importantes contraintes résiduelles sont générées. Leur modélisation est nécessaire pour obtenir des résultats numériques prédictifs. Une première difficulté vient de la modélisation des échanges thermiques. Pour cela, le choix d'une fonction analytique simple pour le coefficient d'échange H = f(Tparoi), suivi de l'optimisation des différents paramètres, a permis de modéliser les courbes de refroidissement. Une fois l'histoire thermique de la culasse modélisée, un calcul mécanique élastoplastique permet de déterminer les contraintes résiduelles. Celles-ci, fortement de traction dans la zone à risque, sont défavorables à la tenue à la fatigue polycyclique de la pièce dans le cas de la trempe à l'eau. La prise en compte de l'état résiduel de fin de traitement thermique dans la filière de dimensionnement à la fatigue polycyclique permet alors de caractériser les zones à risque expérimentalement observées.