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Microsporidia have the leanest genomes among eukaryotes, and their physiological and genomic simplicity has been attributed to their intracellular, obligate parasitic life-style. However, not all microsporidia genomes are small or lean, with the largest dwarfing the smallest ones by at least an order of magnitude. To better understand the evolutionary mechanisms behind this genomic diversification, we explore here two clades of microsporidia with distinct life histories, Ordospora and Hamiltosporidium, parasitizing the same host species, Daphnia magna. Based on seven newly assembled genomes, we show that mixed-mode transmission (the combination of horizontal and vertical transmission), which occurs in Hamiltosporidium, is found to be associated with larger and AT-biased genomes, more genes, and longer intergenic regions, as compared with the exclusively horizontally transmitted Ordospora. Furthermore, the Hamiltosporidium genome assemblies contain a variety of repetitive elements and long segmental duplications. We show that there is an excess of nonsynonymous substitutions in the microsporidia with mixed-mode transmission, which cannot be solely attributed to the lack of recombination, suggesting that bursts of genome size in these microsporidia result primarily from genetic drift. Overall, these findings suggest that the switch from a horizontal-only to a mixed mode of transmission likely produces population bottlenecks in Hamiltosporidium species, therefore reducing the effectiveness of natural selection, and allowing their genomic features to be largely shaped by nonadaptive processes.

Pollinator populations, including bees, are in rapid decline in many parts of the world, raising concerns over the future of ecosystems and food production. Among the factors involved in these declines, poor nutrition deserves attention. The diet consumed by adult worker honeybees (Apis mellifera) is crucial for their behavioral maturation, i.e., the progressive division of labor they perform, such as nurse bees initially and later in life as foragers. Poor pollen nutrition is known to reduce the workers' lifespan, but the underlying physiological and genetic mechanisms are not fully understood. Here we investigate how the lack of pollen in the diet of workers during their first week of adult life can affect age-related phenotypes. During the first seven days of adult life, newly emerged workers were fed either a pollen-deprived (PD) diet mimicking that of an older bee, or a control pollen-rich (PR) diet, as typically consumed by young bees. The PD-fed bees showed alterations in their fat body transcriptome, such as a switch from a protein-lipid based metabolism to a carbohydrate-based metabolism, and a reduced expression of genes involved with immune response. The absence of pollen in the diet also led to an accumulation of oxidative stress markers in fat body tissue and alterations in the cuticular hydrocarbon profiles, which became similar to those of chronologically older bees. Together, our data indicate that the absence of pollen during first week of adulthood triggers the premature onset of an aging-related worker phenotype.

Uterine corpus endometrial carcinoma (UCEC) is the second most common type of gynecological tumor. Several research studies have recently shown the potential of different ncRNAs as biomarkers for prognostics and diagnosis in different types of cancers, including UCEC. Thus, we hypothesized that long noncoding RNAs (lncRNAs) could serve as efficient factors to discriminate solid primary (TP) and normal adjacent (NT) tissues in UCEC with high accuracy. We performed an in silico differential expression analysis comparing TP and NT from a set of samples downloaded from the Cancer Genome Atlas (TCGA) database, targeting highly differentially expressed lncRNAs that could potentially serve as gene expression markers. All analyses were performed in R software. The receiver operator characteristics (ROC) analyses and both supervised and unsupervised machine learning indicated a set of 14 lncRNAs that may serve as biomarkers for UCEC. Functions and putative pathways were assessed through a coexpression network and target enrichment analysis.

International audience; AbstractTo characterize the populational diversity of Mourella caerulea, an endemic stingless bee from the Pampa biome, we collected workers of the stingless bee Mourella caerulea from 24 colonies of five localities in Southern Brazil and analyzed it using geometric morphometrics of forewings, mtDNA cytochrome oxidase I variability, and cuticular hydrocarbon (CHC) chemical analysis. The morphometric analysis discriminated the populations of M. caerulea from different physiographic regions. There was a positive correlation between morphometric and geographic distances. CHC profiles also differentiated the colonies from different localities. We found six particular haplotypes, nucleotide diversity (π) of 0.01631, and a haplotype diversity (Hd) of 0.74. In this sense, the comparison of the population belonging to different physiographic regions indicates that we need to give particular attention to M. caerulea at the moment of creating conservation strategies for South Brazilian Fauna, once it is the only species of this monospecific genus, and its populations are much differentiated from each other.

Medulloblastoma (MB), which originates from embryonic neural stem cells (NSCs) or neural precursors in the developing cerebellum, is the most common malignant brain tumor of childhood. Recurrent and metastatic disease is the principal cause of death and may be related to resistance within cancer stem cells (CSCs). Chromatin state is involved in maintaining signaling pathways related to stemness, and inhibition of histone deacetylase enzymes (HDAC) has emerged as a therapeutic tool to target this cell population. Here, we observed changes in the stemness phenotype induced by HDAC inhibition in MB. Analyses of MB tumor samples showed that the stemness markers BMI1 and CD133 are expressed in all molecular subgroups of MB. We then treated human MB cells with the HDAC inhibitor (HDACi) sodium butyrate (NaB). Increased acetylation mediated by NaB reduced cell viability and expression of BMI1 and CD133. Activation of extracellular-regulated kinase (ERK) signaling was also decreased after treatment with NaB. Enrichment analysis of genes associated with CD133 or BMI1 expression showed mitogen-activated protein kinase (MAPK)/ERK signaling and related pathways as the most enriched processes. We went on to use the mitogen-activated protein kinase kinase (MEK) inhibitor U0126 to explore the role of MAPK/ERK pathway. MEK inhibition reduced expression of the stemness markers, hindered MB neurosphere formation, and its antiproliferative effect was enhanced by combination with NaB. These results suggest that combining HDAC and MAPK/ERK inhibitors may be more effective in reducing MB proliferation than single-drug treatments, through modulation of the stemness phenotype of MB cells.

Made available in DSpace on 2019-10-04T12:36:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-03-14 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Differences in the timing of exoskeleton melanization and sclerotization are evident when comparing eusocial and solitary bees. This cuticular maturation heterochrony may be associated with life style, considering that eusocial bees remain protected inside the nest for many days after emergence, while the solitary bees immediately start outside activities. To address this issue, we characterized gene expression using large-scale RNA sequencing (RNA-seq), and quantified cuticular hydrocarbon (CHC) through gas chromatography-mass spectrometry in comparative studies of the integument (cuticle plus its underlying epidermis) of two eusocial and a solitary bee species. In addition, we used transmission electron microscopy (TEM) for studying the developing cuticle of these and other three bee species also differing in life style. We found 13,200, 55,209 and 30,161 transcript types in the integument of the eusocial Apis mellifera and Frieseomelitta varia, and the solitary Centris analis, respectively. In general, structural cuticle proteins and chitin-related genes were upregulated in pharate-adults and newly-emerged bees whereas transcripts for odorant binding proteins, cytochrome P450 and antioxidant proteins were overrepresented in foragers. Consistent with our hypothesis, a distance correlation analysis based on the differentially expressed genes suggested delayed cuticle maturation in A. mellifera in comparison to the solitary bee. However, this was not confirmed in the comparison with F. varia. The expression profiles of 27 of 119 genes displaying functional attributes related to cuticle formation/differentiation were positively correlated between A. mellifera and F. varia, and negatively or non-correlated with C. analis, suggesting roles in cuticular maturation heterochrony. However, we also found transcript profiles positively correlated between each one of the eusocial species and C. analis. Gene co-expression networks greatly differed between the bee species, but we identified common gene interactions exclusively between the eusocial species. Except for F. varia, the TEM analysis is consistent with cuticle development timing adapted to the social or solitary life style. In support to our hypothesis, the absolute quantities of n-alkanes and unsaturated CHCs were significantly higher in foragers than in the earlier developmental phases of the eusocial bees, but did not discriminate newly-emerged from foragers in C. analis. By highlighting differences in integument gene expression, cuticle ultrastructure, and CHC profiles between eusocial and solitary bees, our data provided insights into the process of heterochronic cuticle maturation associated to the way of life. Univ Sao Paulo, Dept Genet, Fac Med Ribeirao Preto, Ribeirao Preto, Brazil Hosp Clin Porto Alegre, Nucleo Bioinformat, Porto Alegre, RS, Brazil Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Tecnol, Jaboticabal, Brazil Univ Sao Paulo, Dept Biol, Fac Filosofia Ciencias & Letras Ribeirao Preto, Ribeirao Preto, Brazil Univ Sao Paulo, Dept Fis & Quim, Fac Ciencias Farmaceut Ribeirao Preto, Ribeirao Preto, Brazil Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Tecnol, Jaboticabal, Brazil FAPESP: 2014/13136-0 FAPESP: 2011/03171-5 FAPESP: 2014/50265-3 CAPES: 88887.160608/2017-00 CAPES: 001 : FAPESP 2012/24284-5

Differences in the timing of exoskeleton melanization and sclerotization are evident when comparing eusocial and solitary bees. This cuticular maturation heterochrony may be associated with life style, considering that eusocial bees remain protected inside the nest for many days after emergence, while the solitary bees immediately start outside activities. To address this issue, we characterized gene expression using large-scale RNA sequencing (RNA-seq), and quantified cuticular hydrocarbon (CHC) through gas chromatography-mass spectrometry in comparative studies of the integument (cuticle plus its underlying epidermis) of two eusocial and a solitary bee species. In addition, we used transmission electron microscopy (TEM) for studying the developing cuticle of these and other three bee species also differing in life style. We found 13,200, 55,209 and 30,161 transcript types in the integument of the eusocialApis melliferaandFrieseomelitta varia, and the solitaryCentris analis, respectively. In general, structural cuticle proteins and chitin-related genes were upregulated in pharate-adults and newly-emerged bees whereas transcripts for odorant binding proteins, cytochrome P450 and antioxidant proteins were overrepresented in foragers. Consistent with our hypothesis, a distance correlation analysis based on the differentially expressed genes suggested delayed cuticle maturation inA. melliferain comparison to the solitary bee. However, this was not confirmed in the comparison withF. varia. The expression profiles of 27 of 119 genes displaying functional attributes related to cuticle formation/differentiation were positively correlated betweenA. melliferaandF. varia, and negatively or non-correlated withC. analis, suggesting roles in cuticular maturation heterochrony. However, we also found transcript profiles positively correlated between each one of the eusocial species andC. analis. Gene co-expression networks greatly differed between the bee species, but we identified common gene interactions exclusively between the eusocial species. Except forF. varia, the TEM analysis is consistent with cuticle development timing adapted to the social or solitary life style. In support to our hypothesis, the absolute quantities of n-alkanes and unsaturated CHCs were significantly higher in foragers than in the earlier developmental phases of the eusocial bees, but did not discriminate newly-emerged from foragers inC. analis. By highlighting differences in integument gene expression, cuticle ultrastructure, and CHC profiles between eusocial and solitary bees, our data provided insights into the process of heterochronic cuticle maturation associated to the way of life.Author SummaryFrom our previous observation that bees with distinct habits of life, eusocial and solitary, exhibit different degrees of cuticle melanization and sclerotization at the emergence, we decided to analyze the genetic signatures and ultrastructure of the integument, as well as the CHC profiles that could be involved in cuticle maturation. The expression patterns of certain genes involved in the melanization/sclerotization pathway, chitin metabolism, cuticle structure, and also regulators of cuticle renewal and tanning, in addition to other genes, might be grounded the slow process of cuticle maturation in the eusocial bees in comparison to the solitary ones. The electron micrographs revealed differences in the timing of cuticle deposition for the eusocial and solitary species. Among the identified CHCs, the proportions and quantities of n-alkanes in the developing cuticle are consistent with the faster cuticular maturation in the solitary bee, thus supporting our hypothesis.

Lupinus albescens is a resistant cover plant that establishes symbiotic relationships with bacteria belonging to the Bradyrhizobium genus. This symbiosis helps the development of these plants in adverse environmental conditions, such as the ones found in arenized areas of Southern Brazil. This work studied three Bradyrhizobium sp. (AS23, NAS80 and NAS96) isolated from L. albescens plants that grow in extremely poor soils (arenized areas and adjacent grasslands). The genomes of these three strains were sequenced in the Ion Torrent platform using the IonXpress library preparation kit, and presented a total number of bases of 1,230,460,823 for AS23, 1,320,104,022 for NAS80, and 1,236,105,093 for NAS96. The genome comparison with closest strains Bradyrhizobium japonicum USDA6 and Bradyrhizobium diazoefficiens USDA110 showed important variable regions (with less than 80% of similarity). Genes encoding for factors for resistance/tolerance to heavy metal, flagellar motility, response to osmotic and oxidative stresses, heat shock proteins (present only in the three sequenced genomes) could be responsible for the ability of these microorganisms to survive in inhospitable environments. Knowledge about these genomes will provide a foundation for future development of an inoculant bioproduct that should optimize the recovery of degraded soils using cover crops.

The emergence of queens and workers from solitary antecedents mark a major evolutionary transition in the history of life. The solitary progressive provisioning wasp Synagris cornuta, a member of the subfamily Eumeninae (basal to eusocial vespid wasps), alternates between behavioral states characterized as queenlike and worker-like. Akin to a queen in eusocial wasps, a S. cornuta female initiates construction of a cell into which she oviposits and then, similar to a worker, cares for the brood as it develops. The ovarian groundplan (OGP) hypothesis for caste origins predicts that these behavioral states are associated with cyclical changes in ovarian status, where females performing queenlike tasks have eggs and those performing worker-like tasks possess only small oocytes. Our findings show strong support for the OGP hypothesis: the ovaries of S. cornuta females undergo differential oogenesis depending on the behavioral phase: the largest oocyte in the ovaries of females building a cell progresses faster compared to that of females attending brood. Yet contrary to the OGP hypothesis, neither juvenile hormone nor ecdysteroids is associated with the reproductive cycle. Finally, the cuticular hydrocarbon profile showed no link with ovarian status, suggesting that fertility signals evolved subsequent to the emergence of group living.

Hepatocellular carcinoma (HCC) is the prevalent type of primary liver malignancy. Different noncoding RNAs (ncRNAs) that negatively regulate gene expression, such as the microRNAs and the long ncRNAs (lncRNAs), have been associated with cell invasiveness and cell dissemination, tumor recurrence, and metastasis in HCC. To evaluate which regulatory ncRNAs might be good candidates to disrupt HCC proliferation pathways, we performed both unsupervised and supervised analyses of HCC expression data, comparing samples of solid tumor tissue (TP) and adjacent tissue (NT) of a set of patients, focusing on ncRNAs and searching for common mechanisms that may shed light in future therapeutic options. All analyses were performed using the R software. Differential expression (total RNA and miRNA) and enrichment analyses (Gene Ontology + Pathways) were performed using the package TCGABiolinks. As a result, we improved the set of lncRNAs that could be the target of future studies in HCC, highlighting the potential ofFAM170B-AS1andTTN-AS1.

The differentiation of workers into morphological castes represents an important evolutionary innovation that is thought to improve division of labor in insect societies. Given the potential benefits of task-related worker differentiation, it is puzzling that physical worker castes, such as soldiers, are extremely rare in social bees and absent in wasps. Following the recent discovery of soldiers in a stingless bee, we studied the occurrence of worker differentiation in 28 stingless bee species from Brazil and found that several species have specialized soldiers for colony defence. Our results reveal that worker differentiation evolved repeatedly during the last ~ 25 million years and coincided with the emergence of parasitic robber bees, a major threat to many stingless bee species. Furthermore, our data suggest that these robbers are a driving force behind the evolution of worker differentiation as targets of robber bees are four times more likely to have nest guards of increased size than non-targets. These findings reveal unexpected diversity in the social organization of stingless bees., Although common in ants and termites, worker differentiation into physical castes is rare in social bees and unknown in wasps. Here, Grüter and colleagues find a guard caste in ten species of stingless bees and show that the evolution of the guard caste is associated with parasitization by robber bees.

Polyaromatic Hydrocarbons (PAH) are recalcitrant pollutants that are among the major Petroleum by-products. Among them, benzo(a)pyrene receives special attention as it is carcinogenic and highly insoluble, being really difficult to decompose with conventional treatments. Wastewater treatment plants (WWTP), often use biological techniques such as activated sludge, that rely on microorganisms to degrade organic compounds. Burkholderia genus is known for its biodegradation ability and have been used as study model. Using transcripts and mRNA analyses, one can find out genomic regions and functions involved in this aptitude. In this work we used the model strain B. vietnamiensis G4 aiming to find genes that are differentially expressed in the presence of benzo(a)pyrene. Six transcriptomes where generated from each experimental unit in order to compare gene expression and infer which genes are involved in benzo(a)pyrene degradation pathways. Thirty-six genes were differentially expressed in the group exposed to benzo(a)pyrene, most of them involved in catalytic activity. The most significant genomic regions are: phenylacetic acid degradation protein paaN involved in degradation of organic compounds to obtain energy, oxidoreductase FAD-binding subunit in the regulation of electrons within groups of dioxygenase enzymes, with potential to break benzene rings, and glutamic semialdehyde dehydrogenase, a region described as responsible for the ability of phenol degradation. These candidate genes are now target to new studies aiming optimization of treatment processes.

International audience; Epicuticular hydrocarbons are involved in several behavioral processes in social insects including recognition of nestmates, castes, and sex. The aim of the present study was to characterize and to verify differences in the chemical profiles of 1-day recently emerged workers, males, and queens in colonies of the stingless bee Melipona marginata. Gas chromatography–mass spectrometry and multivariate analyses of bee samples from three colonies clearly separated cuticular hydrocarbon profiles of males, queens, and workers. The results showed that cuticular hydrocarbon profiles were comprised of alkanes, alkenes, and methyl alkanes that varied quantitatively and qualitatively according to the castes and sex. Different from previous studies, our investigation also demonstrated that males and queens presented higher similarity in their epicuticular hydrocarbons than to workers.

Os hidrocarbonetos cuticulares têm importante papel no processo de reconhecimento dos membros da colônia de insetos sociais. Muitos estudos têm mostrado variações qualitativas e quantitativas nestes compostos entre os insetos adultos. Contudo, abordagens referentes à modulação do perfil destes compostos durante a formação da cutícula são escassas, e se restringem aos estágios larval de holometábolos e de ninfas de hemimetábolos. O principal objetivo dessa pesquisa foi caracterizar o perfil de hidrocarbonetos cuticulares e a expressão de genes potencialmente relacionados à sua biossíntese durante o processo de formação e maturação da cutícula adulta. Os perfis de hidrocarbonetos foram caracterizados por meio de GC/MS e mostraram diferenças quantitativas marcantes que significativamente discriminaram as cutículas pupal, adulta-farata e adulta. Em paralelo, sequências de enzimas que catalisam a desaturação (desaturases) ou elongação (elongases) de lipídeos, disponíveis no banco de dados do NCBI, foram utilizadas para o desenho de primers e estudo da expressão gênica por meio de RT-qPCR. Cinco genes de desaturases, e oito genes de elongases mostraram variação de expressão estatisticamente significante no tegumento de abelhas adultas em comparação com pupas e adultas-faratas. Testes de correlação entre os perfis de expressão gênica e de hidrocarbonetos cuticulares evidenciaram os genes potencialmente envolvidos com a biossíntese destes compostos para a formação e maturação da cutícula. Estes resultados corroboram a hipótese de que nos insetos sociais, a cutícula só amadurece completamente por ocasião do início da atividade de forrageamento. Associando estes dados a análises de evolução molecular das desaturases e elongases, pudemos sugerir as etapas da via de síntese de hidrocarbonetos catalisadas por estas enzimas, e assim eleger genes candidatos a futuro silenciamento mediado por RNA de interferência para pesquisa de função. Cuticular hydrocarbons are important for recognition of nestmates in social insect colonies. Many studies have shown qualitative and quantitative variations in the cuticular hydrocarbons between adult insects. However, approaches on developmental profiles of these compounds during cuticle formation and differentiation are scarce, and restricted to larval stages of holometabolous and nymphs of hemimetabolous. The main objective of this work was to characterize the cuticular hydrocarbons profiles and the expression of genes potentially involved in the biosynthesis of these compounds during the synthesis and differentiation of the adult cuticle in the honeybee. The hydrocarbons profiles were characterized using GC/MS and showed remarkable quantitative differences, thus discriminating the pupal, pharate-adult and adult cuticles from each other. In parallel, we used annotated sequences of enzymes catalyzing lipid desaturation (desaturases) or elongation (elongases), available in NCBI data bank, for primers design and gene expression analysis using RT-qPCR. Five desaturase genes and eight elongase genes showed statistically significant expression changes in the integument of adult bees in comparison to pupae and pharate-adults. Correlation tests supported roles of some of the desaturase and elongase genes in hydrocarbons biosynthesis for incorporation into adult cuticle. In addition, these results go along with the hypothesis that in social insects the cuticle is just completed when the insect starts forager activity. Taken together, these data and an analysis on the molecular evolution of desaturases and elongases allowed suggesting the steps in the pathway of cuticular hydrocarbons biosynthesis that are catalyzed by these enzymes, and also allowed to elect candidate genes for further functional studies using gene silencing mediated by RNAi.

Made available in DSpace on 2015-10-21T20:31:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-04-24. Added 1 bitstream(s) on 2015-10-22T09:48:01Z : No. of bitstreams: 1 WOS000353676700001.pdf: 2518339 bytes, checksum: d1f8d18bd059b76504f3fcd9aa639a74 (MD5) National Institutes of Health (NIH) Agriculture and Food Research Initiative Competitive grant from the USDA National Institute of Food and Agriculture Research Council of Norway (NFR) PEW Charitable Trust University of East Anglia, UK Israel Science Foundation (ISF) Biotechnology and Biological Sciences Research Council, UK University of East Anglia University of Alabama College of Arts and Sciences Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Marie Curie International Outgoing Fellowship Swiss National Science Foundation New Faculty Initiative Grant (NFIG) from Illinois State University College of Arts and Sciences DFG Instituto Nacional de Bioinformatica (INB) from ISCIII in Spain Background: The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats.Results: We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits.Conclusions: These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation. Illinois State Univ, Sch Biol Sci, Normal, IL 61790 USA ETH, Inst Integrat Biol, Expt Ecol, CH-8092 Zurich, Switzerland E Carolina Univ, Dept Biol, Greenville, NC 27858 USA Hebrew Univ Jerusalem, Alexander Silberman Inst Life Sci, Dept Ecol Evolut &Behav, Jerusalem, Israel Univ Ghent, Fac Sci, Lab Zoophysiol, B-9000 Ghent, Belgium Univ Otago, Dept Biochem, Labo Evolut &Dev Genet, Dunedin 9054, New Zealand Univ Otago, Dept Biochem, Natl Res Ctr Growth &Dev, Dunedin 9054, New Zealand Univ Missouri, Div Plant Sci, Div Anim Sci, Columbia, MO 65211 USA Univ Missouri, MU Informat Inst, Columbia, MO 65211 USA Georgetown Univ, Dept Biol, Washington, DC 20057 USA Arizona State Univ, Sch Life Sci, Tempe, AZ 85287 USA Univ Copenhagen, Dept Biol, Ctr Funct &Comparat Insect Gen, Copenhagen, Denmark Univ Hohenheim, Inst Anim Sci, D-70599 Stuttgart, Germany Univ Alabama, Dept Biol Sci, Tuscaloosa, AL 35487 USA Univ Illinois, Dept Entomol, Urbana, IL 61801 USA Univ Ghent, Fac Biosci Engn, Dept Crop Protect, Lab Agrozool, B-9000 Ghent, Belgium Univ Halle Wittenberg, Inst Biol, Wittenberg, Germany Univ Geneva, Sch Med, Dept Genet Med &Dev, CH-1211 Geneva, Switzerland Swiss Inst Bioinformat, CH-1211 Geneva, Switzerland MIT, Comp Sci &Artificial Intelligence Lab, Cambridge, MA 02139 USA MIT &Harvard, Broad Inst, Cambridge Ctr 7, Cambridge, MA 02142 USA Univ Munster, Inst Evolut &Biodivers, D-48149 Munster, Germany Ctr Genom Regulat CRG, Barcelona 08003, Spain UPF, Barcelona, Spain Ernst Moritz Arndt Univ Greifswald, Inst Math &Comp Sci, D-17487 Greifswald, Germany Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Genom Div, Berkeley, CA 94720 USA Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA Norwegian Univ Food Sci, Dept Chem Biotechnol &Food Sci, N-1432 As, Norway Univ E Anglia, Sch Comp Sci, Norwich NR4 7TJ, Norfolk, England Univ Dusseldorf, Inst Evolut Genet, D-40225 Dusseldorf, Germany Univ Cologne, Inst Genet, Cologne, Germany Univ Sao Paulo, Dept Biol, Fac Filosofia Ciencias &Letras Ribeirao Preto, BR-14040901 Ribeirao Preto, Brazil Univ Edinburgh, Ashworth Labs, Inst Evolutionary Biol &Edinburgh Gen, Edinburgh EH9 3FL, Midlothian, Scotland Univ E Anglia, Sch Biol Sci, Norwich NR4 7TJ, Norfolk, England Royal Holloway Univ London, Sch Biol Sci, London, England Maynooth Univ, Dept Biol, Co, Kildare, Ireland Univ Bristol, Sch Biol Sci, Bristol BS8 1TQ, Avon, England CSIRO, Land &Water Flagship, Canberra, ACT, Australia Trin Coll Dublin, Sch Nat Sci, Dept Zool, Dublin, Ireland Zool Soc London, Inst Zool, London NW1 4RY, England ETH, Inst Integrat Biol, Theoret Biol, CH-8092 Zurich, Switzerland Swiss Inst Bioinformat, Lausanne, Switzerland Swiss Fed Inst Technol, Dept Biosyst Sci &Engn, Computat Evolut, Basel, Switzerland USDA ARS, Bee Res Lab, Washington, DC 20250 USA Univ Sao Paulo, Dept Genet, Fac Med Ribeirao Preto, BR-14040900 Ribeirao Preto, Brazil N Carolina State Univ, Ctr Res Sci Computat, Raleigh, NC 27695 USA Kyoto Univ, Grad Sch Agr, Lab Insect Ecol, Kyoto, Japan Univ Sao Paulo, Dept Biol Celular &Mol &Bioagentes Patogen, Fac Med Ribeirao Preto, BR-14040900 Ribeirao Preto, Brazil Inst Fed Educ Ciencia &Tecnol Sao Paulo, BR-15991502 Matao, Brazil Ohio State Univ, Dept Entomol, Wooster, OH 44791 USA Oxford Brookes Univ, Fac Hlth &Life Sci, Dept Biol &Med Sci, Oxford OX3 0BP, England Xian Jiaotong Liverpool Univ, Dept Biol Sci, Suzhou, Peoples R China Univ Hosp Halle Saale, Dept Lab Med, Halle, Germany German Ctr Integrat Biodivers Res iDiv, Leipzig, Germany Univ Southampton, Southampton, Hants, England Univ Leicester, Dept Biol, Leicester, Leics, England Univ Fed Sao Carlos, Ctr Ciencias Biol &Saude, Dept Genet &Evolucao, BR-13565905 Sao Carlos, SP, Brazil Univ Estadual Paulista, Fac Ciencias Agr &Vet, Dept Tecnol, BR-14884900 Jaboticabal, Brazil Univ N Carolina, Dept Biol, Greensboro, NC 27403 USA Natl Ctr Sci Res Demokritos, Inst Biosci &Applicat, Athens, Greece Univ Munich, Munich, Germany Baylor Coll Med, Dept Mol &Human Genet, Human Genome Sequencing Ctr, Houston, TX 77030 USA Univ Illinois, Roy J Carver Biotechnol, Urbana, IL 61801 USA Univ Illinois, Dept Crop Sci, Urbana, IL 61801 USA Univ Illinois, Inst Genom Biol, Urbana, IL 61801 USA MD Anderson Canc Ctr, Sch Hlth Profess, Mol Genet Technol Program, Unit 2, Houston, TX 77025 USA Univ Chicago, Dept Human Genet, Chicago, IL 60637 USA Univ Ghent, Dept Biochem &Microbiol, Lab Prot Biochem &Biomol Engn, B-9000 Ghent, Belgium Queen Mary Univ London, Sch Biol &Chem Sci, Dept Biol &Expt Psychol, London E1 4NS, England Guangzhou Univ, Sch Life Sci, Guangzhou, Guangdong, Peoples R China Johns Hopkins Univ, McKusick Nathans Inst Genet Med, Ctr Computat Biol, Baltimore, MD 21205 USA Univ Illinois, Neurosci Program, Dept Entomol, Carl R Woese Inst Genom Biol, Urbana, IL 61801 USA Departamento de Tecnologia, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, 14884-900, Brazi National Institutes of Health (NIH): DP1 OD006416 National Institutes of Health (NIH): U54 HG003273 Agriculture and Food Research Initiative Competitive grant from the USDA National Institute of Food and Agriculture: 2010-65106-21301 FAPESP: 11/03171-5 Marie Curie International Outgoing Fellowship: PIOF-GA-2011-303312 Swiss National Science Foundation: 31003A-125350 Swiss National Science Foundation: 31003A-143936

Cuticular hydrocarbons (CHCs) are abundant in the superficial cuticular layer (envelope) of insects where they play roles as structural, anti-desiccation and semiochemical compounds. Many studies have investigated the CHC composition in the adult insects. However, studies on the profiles of these compounds during cuticle formation and differentiation are scarce and restrict to specific stages of a few insect species. We characterized the CHCs developmental profiles in the honeybee workers during an entire molting cycle (from pupal-to-adult ecdyses) and in mature adults (forager bees). Gas chromatography/mass spectrometry (GC/MS) analysis revealed remarkable differences in the relative quantities of CHCs, thus discriminating pupae, developing and newly-ecdysed adults, and foragers from each other. In parallel, the honeybee genome database was searched for predicted gene models using known amino acid sequences of insect enzymes catalyzing lipid desaturation (desaturases) or elongation (elongases) as queries in BLASTP analysis. The expression levels of six desaturase genes and ten elongase genes potentially involved in CHC biosynthesis were determined by reverse transcription and real time polymerase chain reaction (RT-qPCR) in the developing integument (cuticle and subjacent epidermis). Aiming to predict roles for these genes in CHC biosynthesis, the developmental profiles of CHCs and desaturase/elongase transcript levels were evaluated using Spearman correlation coefficient. This analysis pointed to differential roles for these gene products in the biosynthesis of certain CHC classes. Based on the assumption that homologous proteins may share a similar function, phylogenetic trees were reconstructed as an additional strategy to predict functions and evolutionary relationships of the honeybee desaturases and elongases. Together, these approaches highlighted the molecular complexity underlying the formation of the lesser known layer of the cuticular exoskeleton, the envelope.

Context: Reduced longevity observed in hypopituitarism has been attributed to GH deficiency (GHD). It is, however, unclear whether GHD or other confounding factors cause this early mortality. Objective: The aim was to study longevity in subjects from a large kindred with untreated, lifetime isolated GHD (IGHD) due to a homozygous mutation in the GHRH receptor gene and in heterozygous carriers of the mutation. Design, Setting, and Participants: We carried out a retrospective cohort study on three groups. We first compared mortality risk of 65 IGHD individuals and their 128 unaffected siblings from 34 families. We then compared mean age of death of the IGHD to the general population. A transversal study was carried out to compare the rate of heterozygosity for the mutation in two groups of young (20–40 yr old) and old (60–80 yr old) normal-appearing subjects from the same county. Main Outcome Measure: We measured longevity. Results: The risk of death of IGHD subjects was not different from their siblings. Life span in IGHD individuals was shorter than the general population. When stratified by sex, this difference persisted only in females, due to a high frequency of IGHD deaths in females aged 4–20. There was no significant difference in life span between IGHD subjects and siblings or the general population when analyzing subjects who reached age 20. The prevalence of heterozygosity did not differ in young and old groups, suggesting no survival advantage or disadvantage. Conclusions: In a selected genetic background, lifelong untreated IGHD does not affect longevity.