A comprehensive characterization of the caspase gene family in insects from the order Lepidoptera

<p>Abstract</p> <p>Background</p> <p>The cell suicide pathway of apoptosis is a necessary event in the life of multicellular organisms. It is involved in many biological processes ranging from development to the immune response. Evolutionarily conserved proteases, called caspases, play a central role in regulating apoptosis. Reception of death stimuli triggers the activation of initiator caspases, which in turn activate the effector caspases. In Lepidoptera, apoptosis is crucial in processes such as metamorphosis or defending against baculovirus infection. The discovery of p35, a baculovirus protein inhibiting caspase activity, has led to the characterization of the first lepidopteran caspase, Sf-Caspase-1. Studies on Sf-Caspase-1 mode of activation suggested that apoptosis in Lepidoptera requires a cascade of caspase activation, as demonstrated in many other species.</p> <p>Results</p> <p>In order to get insights into this gene family in Lepidoptera, we performed an extensive survey of lepidopteran-derived EST datasets. We identified 66 sequences distributed among 27 species encoding putative caspases. Phylogenetic analyses showed that Lepidoptera possess at least 5 caspases, for which we propose a unified nomenclature. According to homology to their <it>Drosophila </it>counterparts and their primary structure, we determined that Lep-Caspase-1, -2 and -3 are putative effector caspases, whereas Lep-Caspase-5 and -6 are putative initiators. The likely function of Lep-Caspase-4 remains unclear. Lep-Caspase-2 is absent from the silkworm genome and appears to be noctuid-specific, and to have arisen from a tandem duplication of the Caspase-1 gene. In the tobacco hawkmoth, 3 distinct transcripts encoding putative Caspase-4 were identified, suggesting at least 2 duplication events in this species.</p> <p>Conclusions</p> <p>The basic repertoire of five major types of caspases shared among Lepidoptera seems to be smaller than for most other groups studied to date, but gene duplication still plays a role in lineage-specific increases in diversity, just as in Diptera and mammals.</p

Characterization of Caspases from Lepidoptera and their role in Apoptosis

Multicellular organisms need a tightly controlled regulation of cell proliferation and of cell specialization together with a controlled cell death to maintain their integrity. A family of evolutionarily conserved proteases, called caspases, plays a central role in regulating apoptosis in animals. The discovery of p35, a baculovirus caspase inhibitor, has led to the characterization of the first lepidopteran caspase, Sf-Caspase-1, from Spodoptera frugiperda. Studies on the Sf-Caspase-1 mode of activation suggested that apoptosis in Lepidoptera also requires a cascade of caspase activation, as demonstrated in many other species. We identified 66 sequences encoding putative caspases, distributed among 27 species. Phylogenetic analyses showed that Lepidoptera possess at least 5 caspases, for which we propose a unified nomenclature. According to their homology to their Drosophila counterparts and their primary structure, we proposed that Lep-Caspase-1, -2 and -3 are effector caspases, whereas Lep-Caspase-5 and -6 are putative initiators. The likely function of Lep-Caspase-4 remains unclear. We found that Caspase-1 and -2 have complementary expression profiles during larval development, suggesting a differential regulation. Caspase-3 and -6 are upregulated upon immune challenge, suggesting a role in the imd pathway. Caspase-5 is upregulated during pupation and upon induction with 20-hydroxyecdysone, supporting the hypothesis of Caspase-5 playing a similar role as Dronc in developmental apoptosis in Drosophila. In an attempt to obtain further insights into the molecular pathway underlying apoptosis in lepidopteran insects, we used 2D DIGE to identify 13 proteins, for which relative abundance was altered during apoptosis of H. armigera derived cells, induced by actinomycin D. Among these 13 proteins, we identified the putative effector Caspase-1, 3 chaperone proteins and several pro-apoptotic proteins, such as actin and a voltage dependant anion channel

Modelling habitat sustability for some breeding birds in agricultural landscapes of south west France. A case study

National audienc

Development of heart-cutting multidimensional gas chromatography coupled to time of flight mass spectrometry for silicon speciation at trace levels in gasoline samples

cited By 8International audienceTo improve the understanding of hydrotreatment (HDT) catalyst poisoning by silicon species, these molecules must be characterized in petroleum products using powerful analytical systems. Heart-cutting gas chromatography coupled to time of flight mass spectrometry (GC-GC/TOFMS) method equipped with a Deans switch (DS) system was developed for the direct characterization of target silicon compounds at trace level (μgkg-1) in gasoline samples. This method was performed to identify silicon compounds never characterized before. After the selection of the second dimension column using GC-GC-FID, GC-GC/TOFMS was performed. The calibration curves obtained by the GC-GC/TOFMS method were linear up to 1000μgkg-1. Limits of detection (LOD) were ranging from 5 to 33μgkg-1 in spiked gasoline. The method provided sufficient selectivity and sensitivity to characterize known silicon compounds thanks to their specific ions and their retention times. The analysis of a naphtha sample by GC-GC/TOFMS has shown the presence of cyclic siloxanes (Dn) as major compounds of PDMS thermal degradation with the occurrence of linear siloxanes, especially hexamethyldisiloxane (L2), which was never characterized in petroleum products but already known as severe poison for catalyst. © 2012 Elsevier B.V

Silicon speciation by gas chromatography coupled to mass spectrometry in gasolines

cited By 14International audienceA method for the speciation of silicon compounds in petroleum products was developed using gas chromatography coupled to mass spectrometry (GC-MS). Prior to analysis, several precautions about storage and conservation were applied for all samples. In spiked gasoline samples, limits of detection between 24 and 69μgkg -1 for cyclic siloxanes (D 4-D 6) and between 1 and 7μgkg -1 for other species were obtained. In this study, cyclic siloxanes (D n) and one ethoxysilane were quantified for the first time in petroleum products by a specific method based on response factor calculation to an internal standard. This method was applied to four samples of naphthas and gasolines obtained from a steam cracking process. Cyclic siloxanes were predominant in four investigated samples with concentrations ranging between 101 and 2204μgkg -1. Cyclic siloxane content decreased with an increase in their degree of polymerization. During a steam cracking process, silicon concentrations determined by GC-MS SIM (single ion monitoring) significantly increase. This trend was confirmed by ICP-OES (inductively coupled plasma optical emission spectroscopy) measurements but a difference on the total silicon content was observed, certainly highlighting the presence of unknown silicon species. GC-MS SIM method gives access to the chemical nature of the silicon species, which is crucial for the understanding of hydrotreatment catalyst poisoning in the oil and gas industry. © 2011 Elsevier B.V

Combining fourier transform-ion cyclotron resonance/mass spectrometry analysis and kendrick plots for silicon speciation and molecular characterization in petroleum products at trace levels

cited By 13International audienceA new method combining FT-ICR/MS analysis and Kendrick plots for the characterization of silicon species at trace levels in light petroleum products is presented. The method provides efficient instrumental detection limits ranging from 80 ng/kg to 5 μg/kg and reliable mass accuracy lower than 0.50 ppm for model silicon molecules in spiked gasoline. More than 3000 peaks could be detected in the m/z 50-500 range depending on the nature of the gasoline sample analyzed. An in-house software program was used to calculate Kendrick plots. Then, an algorithm searched, selected, and represented silicon species classes (O 2Si, O 3Si, and O 4Si classes) in Kendrick plots by incorporating model molecules' information (i.e., exact mass and intensity). This procedure allowed the complete characterization of more than 50 new silicon species with different degrees of unsaturation in petroleum products. © 2012 American Chemical Society

Characterization of silicon species issued from PDMS degradation under thermal cracking of hydrocarbons: Part 2 - Liquid samples analysis by a multi-technical approach based on gas chromatography and mass spectrometry

cited By 5International audienceSilicon speciation has recently gained interest in the oil and gas industry due to the impact of silicon species on hydrotreatment (HDT) catalysts. The determination of the chemical structure of silicon compounds appears as essential to limit silicon poisoning as to improve the lifetime of catalysts. To achieve a representative speciation of silicon in petroleum products, fresh samples of PDMS degradation under thermal cracking of hydrocarbons were produced using a pilot plant. The samples were carefully stored in a dewar containing liquid nitrogen (-195 C) to minimize the possible evolution of silicon species and to allow their analysis in representative conditions. A complete analytical approach based on gas chromatography (GC) and mass spectrometry techniques (MS) was developed and applied to the samples produced at 500 C. Moreover, to resolve coelutions observed by GC/TOFMS, GC-GC/TOFMS was successfully applied to obtain the mass spectrum of only one silicon compound. Combining the GC/MS mass spectrum giving access to the fragmentation of the compound and the raw formula and double bond equivalent (DBE) obtained by ESI-FT-ICR/MS, chemical structures were proposed. Almost molecules were strengthened by MSn. Cyclic siloxanes (Dn) were confirmed as the major compounds of PDMS thermal degradation even in the presence of hydrocarbons with a relative amount generally around 95% or above. No significant difference on the formation of Dn were observed according to the different operating conditions under thermal cracking of hydrocarbons. For the first time, several other silicon compounds present at trace levels (&lt;5% of the total area) were characterized. α,ω-dihydroxy polydimethylsiloxanes, methylhydroxy cyclic siloxanes, (n + 1)oxasilabicyclo alcanes or bis(cyclosiloxanyl) siloxane, α-hydroxy, ω-methyl polysiloxanes and (n)oxasilabicyclo alcanes were preferentially formed in the presence of steam. Under evaluated coking or visbreaking conditions (long residence time without steam), other compounds were mainly characterized such as dimethoxy polysiloxanes, methyl(hydroperoxy) cyclic siloxanes, or ethoxy methyl cyclic siloxanes, linear polysiloxanes and methylpropyl cyclic siloxanes. Around one hundred silicon compounds were highlighted belonging to 12 different chemical families. The same repetition pattern (C2H6OSi), initially in PDMS, was present in all silicon compounds characterized in this study. Molecules with a number of silicon atoms ranging from 1 to 40 silicon atoms clearly demonstrated the occurrence of silicon in all petroleum cuts through very different chemical structures. This study shows that silicon species can distillate from gas fractions to the heavy petroleum cuts depending on their boiling points and chemical properties. These silicon species contain reactive groups (hydroxy, hydroperoxy, methoxy and ethoxy) which are able to react at the surface of HDT catalysts and cause a severe deactivation. Thus, these results appear as a crucial advance in progressing in the understanding of silicon poisoning. © 2013 Elsevier Ltd. All rights reserved

Characterization of silicon species issued from PDMS degradation under thermal cracking of hydrocarbons: Part 1 - Gas samples analysis by gas chromatography-time of flight mass spectrometry

cited By 3International audienceSilicon species are becoming emergent contaminants in the oil and gas industry due to their severe poisoning effect on the hydrotreatment (HDT) catalysts. Using an experimental pilot plant, fresh and representative samples of PDMS degradation under thermal cracking of hydrocarbons were produced. To follow the evolution of silicon species, the gas fraction was immediately analyzed by GC/TOFMS after the production and also after 4 months of storage at 4 C. Cyclic siloxanes (Dn) as the major products of PDMS thermal degradation were characterized in the gas phase but these compounds are mainly present in the liquid fraction. Five volatile silicon compounds belonging to the families of silanes, siloxanes and silanols were characterized and quantified in the thermal cracking samples depending on the operating conditions applied in degradation tests. Under coking or visbreaking conditions (long residence time, absence of steam), silanes and siloxanes were preferentially formed. Under evaluated steam cracking conditions (short residence time and presence of steam), trimethylsilanol (TMSOH) was mainly produced by the hydrolysis of PDMS. The formation of the linear siloxane (L2) after several month of storage at 4 C by the self-condensation of TMSOH was also observed. The suspected poisoning effects of these molecules were discussed and could explain the deactivation of catalysts taking place in the refining of the light petroleum cuts. The new identified volatile silicon compounds could affect the performance of the catalyst by the reaction of hydroxyl groups potentially present at the surface of the support with reactive silicon molecules, more specifically silanols. © 2013 Elsevier Ltd. All rights reserved