A highly sensitive high-performance liquid chromatography-mass spectrometry method for quantification of fludarabine triphosphate in leukemic cells

A high-performance liquid chromatography (HPLC)-mass spectrometry (MS) method has been developed for the analysis of 9-beta-D-arabinofuranosyl-2-fluoroadenine 5'-triphosphate (F-ara-ATP) from biological samples. Quantification is carried out by selected ion monitoring of the parent ion. Baseline separation of the monophosphate (F-ara-AMP) and diphosphate (F-ara-ADP) is achieved using the volatile ion-pairing reagent dimethylhexylamine. This method is selective and sensitive with an on-column detection limit of approximately 50 fmol. It also permits simultaneous monitoring of endogenous adenosine phosphates. The utility of the assay has been demonstrated by the analysis of F-ara-ATP in human leukemic cells after incubation with 9-beta-D-arabinosyl-2-fluoroadenine (F-ara-A) at clinically relevant concentrations.     

Buprenorphine does not affect acute murine toxoplasmosis and is recommended as an analgesic in Toxoplasma gondii studies in mice

Groups of mice were infected with tachyzoites of the RH strain of Toxoplasma gondii, treated with the opioid analgesic buprenorphine, sodium sulfadiazine, a combination of buprenorphine and sodium sulfadiazine, or nothing in the drinking water, on days -1 to 12 postinfection. Mice in the T. gondii-infected buprenorphine-treated group did not live significantly longer (P > 0.05) than mice given T. gondii and not treated with buprenorphine. Clinical observations of mice indicated that buprenorphine treatment reduced distress and pain in mice with acute toxoplasmosis. Mice treated with sodium sulfadiazine alone or sodium sulfadiazine combined with buprenorphine survived the 28-day study. Mice treated with buprenorphine and not infected with T. gondii also survived the 28 days. This study demonstrates that buprenorphine does not adversely interfere with acute T. gondii infection and indicates that buprenorphine can be given to mice to alleviate pain and distress associated with a T. gondii infection, and not adversely influence the results of toxoplasmosis studies. Analgesic (buprenorphine) treatment should now be the standard of care for mice in acute toxoplasmosis studies.     

Metabolism of fatty acid in yeast: characterisation of beta-oxidation and ultrastructural changes in the genus Sporidiobolus sp. cultivated on ricinoleic acid methyl ester

Cell structure modifications and beta-oxidation induction were monitored in two strains of Sporidiobolus, Sp. Ruinenii and Sp. pararoseus after cultivation on ricinoleic acid methyl ester. Ultrastructural observations of the yeast before and after cultivation on fatty acid esters did not reveal major modifications in Sp. ruinenii. Unexpectedly, in Sp. pararoseus a proliferation of the mitochondrion was observed. After induction, Sp. ruinenii principally exhibited an increase in the activities of acyl-CoA oxidase (ACO), hydroxyacyl-CoA deshydrogenase (HAD), thiolase and catalase. In contrast, Sp. pararoseus lacked ACO and catalase activities, but an increase in acyl-CoA deshydrogenase (ACDH) and enoyl-CoA hydratase (ECH) activity was observed. These data suggest that in Sp. ruinenii, beta-oxidation is preferentially localized in the microbody, whereas in Sp. pararoseus it might be localized in the mitochondria.     

Genome-wide analysis of membrane targeting by S. cerevisiae pleckstrin homology domains

Pleckstrin homology (PH) domains are small protein modules known for their ability to bind phosphoinositides and to drive membrane recruitment of their host proteins. We investigated phosphoinositide binding (in vitro and in vivo) and subcellular localization, and we modeled the electrostatic properties for all 33 PH domains encoded in the S. cerevisiae genome. Only one PH domain (from Num1p) binds phosphoinositides with high affinity and specificity. Six bind phosphoinositides with moderate affinity and little specificity and are membrane targeted in a phosphoinositide-dependent manner. Although all of the remaining 26 yeast PH domains bind phosphoinositides very weakly or not at all, three were nonetheless efficiently membrane targeted. Our proteome-wide analysis argues that membrane targeting is important for only approximately 30% of yeast PH domains and is defined by binding to both phosphoinositides and other targets. These findings have significant implications for understanding the function of proteins that contain this common domain.     

Influence des traitements phytosanitaires sur les biocoenoses du phytoplancton limnique

We used specific diversity of phytoplankton and chlorophyll concentrations as a measure of algal biomass in ponds located in agricultural areas where pesticides and fertilizers are currently used and in natural meadows where pesticides spraying has never occurred.A comparison of the biocoenotic structure of the primary producers in the two types of ponds shows conspicuous differences. Species richness is higher in meadow ponds (25 species) than in agricultural area ponds (10 species). The chlorophyll levels are reduced in agricultural area ponds. These effects could be ascribed to contamination by pesticides.

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Influence des traitements phytosanitaires sur les biocoenoses du phytoplancton limnique

     

The Kinesin-3, Unc-104 Regulates Dendrite Morphogenesis and Synaptic Development in Drosophila

Kinesin-based transport is important for synaptogenesis, neuroplasticity, and maintaining synaptic function. In an anatomical screen of neurodevelopmental mutants, we identified the exchange of a conserved residue (R561H) in the forkhead-associated domain of the kinesin-3 family member Unc-104/KIF1A as the genetic cause for defects in synaptic terminal- and dendrite morphogenesis. Previous structure-based analysis suggested that the corresponding residue in KIF1A might be involved in stabilizing the activated state of kinesin-3 dimers. Herein we provide the first in vivo evidence for the functional importance of R561. The R561H allele (unc-104^bris) is not embryonic lethal, which allowed us to investigate consequences of disturbed Unc-104 function on postembryonic synapse development and larval behavior. We demonstrate that Unc-104 regulates the reliable apposition of active zones and postsynaptic densities, possibly by controlling site-specific delivery of its cargo. Next, we identified a role for Unc-104 in restraining neuromuscular junction growth and coordinating dendrite branch morphogenesis, suggesting that Unc-104 is also involved in dendritic transport. Mutations in KIF1A/unc-104 have been associated with hereditary spastic paraplegia and hereditary sensory and autonomic neuropathy type 2. However, we did not observe synapse retraction or dystonic posterior paralysis. Overall, our study demonstrates the specificity of defects caused by selective impairments of distinct molecular motors and highlights the critical importance of Unc-104 for the maturation of neuronal structures during embryonic development, larval synaptic terminal outgrowth, and dendrite morphogenesis.     

Gene flow and population structure of a solitary top carnivore in a human‐dominated landscape

While African leopard populations are considered to be continuous as demonstrated by their high genetic variation, the southernmost leopard population exists in the Eastern and Western Cape, South Africa, where anthropogenic activities may be affecting this population's structure. Little is known about the elusive, last free‐roaming top predator in the region and this study is the first to report on leopard population structuring using nuclear DNA. By analyzing 14 microsatellite markers from 40 leopard tissue samples, we aimed to understand the populations' structure, genetic distance, and gene flow (Nm). Our results, based on spatially explicit analysis with Bayesian methods, indicate that leopards in the region exist in a fragmented population structure with lower than expected genetic diversity. Three population groups were identified, between which low to moderate levels of gene flow were observed (Nm 0.5 to 3.6). One subpopulation exhibited low genetic differentiation, suggesting a continuous population structure, while the remaining two appear to be less connected, with low emigration and immigration between these populations. Therefore, genetic barriers are present between the subpopulations, and while leopards in the study region may function as a metapopulation, anthropogenic activities threaten to decrease habitat and movement further. Our results indicate that the leopard population may become isolated within a few generations and suggest that management actions should aim to increase habitat connectivity and reduce human–carnivore conflict. Understanding genetic diversity and connectivity of populations has important conservation implications that can highlight management of priority populations to reverse the effects of human‐caused extinctions.     

Within-Crop Air Temperature and Humidity Outcomes on Spatio-Temporal Distribution of the Key Rose Pest Frankliniella occidentalis

Frankliniella occidentalis (Pergande) is a key pest of various crops worldwide. In this study, we analyse the dependence of the infestation of this pest on spatially distributed micro climatic factors in a rose greenhouse. Despite the importance of this subject, the few existing studies have been realized in laboratory rather than in greenhouse conditions. However, recent progress on greenhouse microclimate characterisation has highlighted the strong indoor climate heterogeneity that may influence the within-crop pest distribution. In this study, both microclimate (air temperature and humidity) and thrips distribution were simultaneously mapped in a rose greenhouse. The measurements were sensed in a horizontal plane situated at mid-height of the rose crop inside the greenhouse. Simultaneously, thrips population dynamics were assessed after an artificial and homogeneous infestation of the rose crop. The spatio-temporal distribution of climate and thrips within the greenhouse were compared, and links between thrips infestation and climatic conditions were investigated. A statistical model was used to define the favourable climate conditions for thrips adults and larvae. Our results showed that (i) the air temperature and air humidity were very heterogeneously distributed within the crop, (ii) pest populations aggregated in the most favourable climatic areas and (iii) the highest population density of thrips adults and larvae were recorded at 27°C and 22°C for temperature and 63% and 86% for humidity, respectively. These findings confirm, in real rose cropping conditions, previous laboratory studies on the F. occidentalis climatic optimum and provide a solid scientific support for climatic-based control methods against this pest.     

FORMIDABEL: The Belgian Ants Database

FORMIDABEL is a database of Belgian Ants containing more than 27.000 occurrence records. These records originate from collections, field sampling and literature. The database gives information on 76 native and 9 introduced ant species found in Belgium. The collection records originated mainly from the ants collection in Royal Belgian Institute of Natural Sciences (RBINS), the ‘Gaspar’ Ants collection in Gembloux and the zoological collection of the University of Liège (ULG). The oldest occurrences date back from May 1866, the most recent refer to August 2012. FORMIDABEL is a work in progress and the database is updated twice a year.The latest version of the dataset is publicly and freely accessible through this url: http://ipt.biodiversity.be/resource.do?r=formidabel. The dataset is also retrievable via the GBIF data portal through this link: http://data.gbif.org/datasets/resource/14697A dedicated geo-portal, developed by the Belgian Biodiversity Platform is accessible at: http://www.formicidae-atlas.bePurpose: FORMIDABEL is a joint cooperation of the Flemish ants working group “Polyergus” (http://formicidae.be) and the Wallonian ants working group “FourmisWalBru” (http://fourmiswalbru.be). The original database was created in 2002 in the context of the preliminary red data book of Flemish Ants (Dekoninck et al. 2003). Later, in 2005, data from the Southern part of Belgium; Wallonia and Brussels were added. In 2012 this dataset was again updated for the creation of the first Belgian Ants Atlas (Figure 1) (Dekoninck et al. 2012). The main purpose of this atlas was to generate maps for all outdoor-living ant species in Belgium using an overlay of the standard Belgian ecoregions. By using this overlay for most species, we can discern a clear and often restricted distribution pattern in Belgium, mainly based on vegetation and soil types.Open in a separate windowFigure 1.www.formicidae-atlas.be