RESUMEN
The EU General Data Protection Regulation (GDPR) requirements have prompted a shift from centralised controlled access genome-phenome archives to federated models for sharing sensitive human data. In a data-sharing federation, a central node facilitates data discovery; meanwhile, distributed nodes are responsible for handling data access requests, concluding agreements with data users and providing secure access to the data. Research institutions that want to become part of such federations often lack the resources to set up the required controlled access processes. The DS-PACK tool assembly is a reusable, open-source middleware solution that semi-automates controlled access processes end-to-end, from data submission to access. Data protection principles are engraved into all components of the DS-PACK assembly. DS-PACK centralises access control management and distributes access control enforcement with support for data access via cloud-based applications. DS-PACK is in production use at the ELIXIR Luxembourg data hosting platform, combined with an operational model including legal facilitation and data stewardship.
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Difusión de la Información , Humanos , Acceso a la Información , Seguridad Computacional , Programas InformáticosRESUMEN
Arms race dynamics are a common outcome of host-parasite coevolution. While they can theoretically be maintained indefinitely, realistic arms races are expected to be finite. Once an arms race has ended, for example due to the evolution of a generalist-resistant host, the system may transition into coevolutionary dynamics that favour long-term diversity. In microbial experiments, host-parasite arms races often transition into a stable coexistence of generalist-resistant hosts, (semi-)susceptible hosts, and parasites. While long-term host diversity is implicit in these cases, parasite diversity is usually overlooked. In this study, we examined parasite diversity after the end of an experimental arms race between a unicellular alga (Chlorella variabilis) and its lytic virus (PBCV-1). First, we isolated virus genotypes from multiple time points from two replicate microcosms. A time-shift experiment confirmed that the virus isolates had escalating host ranges, i.e., that arms races had occurred. We then examined the phenotypic and genetic diversity of virus isolates from the post-arms race phase. Post-arms race virus isolates had diverse host ranges, survival probabilities, and growth rates; they also clustered into distinct genetic groups. Importantly, host range diversity was maintained throughout the post-arms race phase, and the frequency of host range phenotypes fluctuated over time. We hypothesize that this dynamic polymorphism was maintained by a combination of fluctuating selection and demographic stochasticity. Together with previous work in prokaryotic systems, our results link experimental observations of arms races to natural observations of long-term host and parasite diversity.
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Chlorella , Chlorella/virología , Chlorella/genética , Variación Genética , Coevolución Biológica , Evolución BiológicaRESUMEN
The present investigation accounts for the environmental impact assessment of an intense algal bloom caused by the dinoflagellate Noctilucas scintillans. The bloom was first observed on the 10th of September 2019, in the vicinity of the Mandapam group of Islands, spreading from Rameswaram Coast in the North (9° 14' 15â³ N, 79° 9' 46â³ E) to Hare Island in the South (9° 14' 51â³ N, 79° 5' 48â³ E). The coastal waters in and around the Mandapam region appeared dark green, and the microscopic examination of the water sample revealed the presence of N. scintillans in large numbers. N. scintillans is a bioluminescent organism; it is inflated and sub-spherical in shape, and the size of the organisms ranged from 350 to 1300 microns. During the intense periods of the bloom, the average density of N. scintillans was recorded with 226.5 × 103 cells/l, and the dissolved oxygen content was very low and the ammonia content was extremely high in certain sites (avg. 4.3635 µm/l). Intensive bloom may lead to a loss of biodiversity in the affected areas of the region. Subsequent investigations indicated that the resilience of the ecosystem in response to natural adversity.
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Dinoflagelados , Fitoplancton , Ecosistema , Amoníaco , Islas , Monitoreo del Ambiente , Dinoflagelados/fisiología , Eutrofización , Agua , Oxígeno , IndiaRESUMEN
OBJECTIVES: Nigeria reported an upsurge in cholera cases in October 2020, which then transitioned into a large, disseminated epidemic for most of 2021. This study aimed to describe the epidemiology, diagnostic performance of rapid diagnostic test (RDT) kits and the factors associated with mortality during the epidemic. DESIGN: A retrospective analysis of national surveillance data. SETTING: 33 of 37 states (including the Federal Capital Territory) in Nigeria. PARTICIPANTS: Persons who met cholera case definition (a person of any age with acute watery diarrhoea, with or without vomiting) between October 2020 and October 2021 within the Nigeria Centre for Disease Control surveillance data. OUTCOME MEASURES: Attack rate (AR; per 100 000 persons), case fatality rate (CFR; %) and accuracy of RDT performance compared with culture using area under the receiver operating characteristic curve (AUROC). Additionally, individual factors associated with cholera deaths and hospitalisation were presented as adjusted OR with 95% CIs. RESULTS: Overall, 93 598 cholera cases and 3298 deaths (CFR: 3.5%) were reported across 33 of 37 states in Nigeria within the study period. The proportions of cholera cases were higher in men aged 5-14 years and women aged 25-44 years. The overall AR was 46.5 per 100 000 persons. The North-West region recorded the highest AR with 102 per 100 000. Older age, male gender, residency in the North-Central region and severe dehydration significantly increased the odds of cholera deaths. The cholera RDT had excellent diagnostic accuracy (AUROC=0.91; 95% CI 0.87 to 0.96). CONCLUSIONS: Cholera remains a serious public health threat in Nigeria with a high mortality rate. Thus, we recommend making RDT kits more widely accessible for improved surveillance and prompt case management across the country.
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Cólera , Epidemias , Cólera/diagnóstico , Cólera/epidemiología , Diarrea/epidemiología , Brotes de Enfermedades , Femenino , Humanos , Masculino , Nigeria/epidemiología , Juego de Reactivos para Diagnóstico , Estudios RetrospectivosRESUMEN
An increasing number of small and medium-sized enterprises (SMEs) in the German organic agri-food sector involves citizens through different community financing models. While such models provide alternative funding sources as well as marketing opportunities to SMEs, they allow private investors to combine their financial and ethical concerns by directly supporting the development of a more sustainable food system. Due to the low level of financial intermediation, community financing is characterized by close relations between investors and investees. Against this background, we apply the proximity concept from economic geography to explore spatial and relational aspects of community financing in the German organic agri-food sector. Based on a qualitative multiple case study approach, we find that the relevance of proximity is twofold. While different forms of proximity between SMEs and their potential investors are key success factors, proximity is also considered as one desired outcome of community financing. Furthermore, our results reveal that the extent to which SMEs rely on particular proximity dimensions distinguishes two different approaches to community financing.
RESUMEN
To capture the global gene network regulating the differentiation of immature T cells in an unbiased manner, large-scale forward genetic screens in zebrafish were conducted and combined with genetic interaction analysis. After ENU mutagenesis, genetic lesions associated with failure of T cell development were identified by meiotic recombination mapping, positional cloning, and whole genome sequencing. Recessive genetic variants in 33 genes were identified and confirmed as causative by additional experiments. The mutations affected T cell development but did not perturb the development of an unrelated cell type, growth hormone-expressing somatotrophs, providing an important measure of cell-type specificity of the genetic variants. The structure of the genetic network encompassing the identified components was established by a subsequent genetic interaction analysis, which identified many instances of positive (alleviating) and negative (synthetic) genetic interactions. Several examples of synthetic lethality were subsequently phenocopied using combinations of small molecule inhibitors. These drugs not only interfered with normal T cell development, but also elicited remission in a model of T cell acute lymphoblastic leukaemia. Our findings illustrate how genetic interaction data obtained in the context of entire organisms can be exploited for targeted interference with specific cell types and their malignant derivatives.
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Redes Reguladoras de Genes , Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Mutaciones Letales Sintéticas , Linfocitos T/metabolismo , Animales , Modelos Animales de Enfermedad , Epistasis Genética , Fenotipo , Pez CebraRESUMEN
Paratuberculosis is a major disease in cattle that severely affects animal welfare and causes huge economic losses worldwide. Development of alternative diagnostic methods is of urgent need to control the disease. Recent studies suggest that long non-coding RNAs (lncRNAs) play a crucial role in regulating immune function and may confer valuable information about the disease. However, their role has not yet been investigated in cattle with respect to infection towards Paratuberculosis. Therefore, we investigated the alteration in genomic expression profiles of mRNA and lncRNA in bovine macrophages in response to Paratuberculosis infection using RNA-Seq. We identified 397 potentially novel lncRNA candidates in macrophages of which 38 were differentially regulated by the infection. A total of 820 coding genes were also significantly altered by the infection. Co-expression analysis of lncRNAs and their neighbouring coding genes suggest regulatory functions of lncRNAs in pathways related to immune response. For example, this included protein coding genes such as TNIP3, TNFAIP3 and NF-κB2 that play a role in NF-κB2 signalling, a pathway associated with immune response. This study advances our understanding of lncRNA roles during Paratuberculosis infection.
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Enfermedades de los Bovinos/genética , Enfermedades de los Bovinos/microbiología , Macrófagos/metabolismo , Mycobacterium avium subsp. paratuberculosis/fisiología , Paratuberculosis/genética , Paratuberculosis/microbiología , ARN Largo no Codificante , ARN Mensajero , Animales , Bovinos , Enfermedades de los Bovinos/inmunología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Genómica/métodos , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Macrófagos/inmunología , Paratuberculosis/inmunología , Interferencia de ARN , Reproducibilidad de los Resultados , Transcripción Genética , TranscriptomaRESUMEN
Insulin-like growth factors (IGFs) play a critical role in fetal growth, and components of the IGF system have been associated with fetal growth restriction in women. In human pregnancy, the proteolytic cleavage of insulin-like growth factor binding proteins (IGFBPs), particularly IGFBP-4, releases free IGF for respective action at the tissue level. The aim of the present study was to determine IGFBP-2, IGFBP-3, and IGFBP-4 concentrations by Western ligand blotting during pregnancy until day 100 in cows and to compare these concentrations with those of non-pregnant cows and cows undergoing embryonic/fetal mortality. Therefore, two study trials (I and II) and an in vitro study were conducted. In study I, 43 cows were not pregnant, 34 cows were pregnant, and 4 cows were undergoing fm. In study II, 500 cows were examined, and 7 cases of pregnancy loss between days 24-27 and 34-37 after artificial insemination (AI, late embryonic mortality; em) and 8 cases of pregnancy loss between days 34-37 and 54-57 after AI (late embryonic mortality and early fetal mortality; em/fm) were defined from the analyses of 30 pregnant and 20 non-pregnant cows randomly selected for insulin-like growth factor 1 and IGFBP analyses. In vitro serum from pregnant (n = 3) and non-pregnant (n = 3) cows spiked after incubation with recombinant human (rh) IGFBP-4 for 24 h, and IGFBP-4 levels were analyzed before and after incubation to detect proteolytic degradation. The IGFBP-2, -3, and -4 concentrations did not decline during early pregnancy in cows, while IGFBP-4 concentrations were comparable between pregnant and non-pregnant cows, irrespective of low proteolytic activity, which was also demonstrated in cows. Interestingly, cows with em or fm showed distinct IGFBP patterns. The IGFBP-2 and -3 concentrations were higher (P < 0.05) in cows with fm compared to pregnant. The IGFBP-4 levels were significantly higher in cows developing fm. Thus, distinct differences in the circulating IGFBP concentrations could be associated with late embryonic and early fetal losses in cattle.
RESUMEN
Cytidine deaminases of the AID/APOBEC family catalyze C-to-U nucleotide transitions in mRNA or DNA. Members of the APOBEC3 branch are involved in antiviral defense, whereas AID contributes to diversification of antibody repertoires in jawed vertebrates via somatic hypermutation, gene conversion, and class switch recombination. In the extant jawless vertebrate, the lamprey, two members of the AID/APOBEC family are implicated in the generation of somatic diversity of the variable lymphocyte receptors (VLRs). Expression studies linked CDA1 and CDA2 genes to the assembly of VLRA/C genes in T-like cells and the VLRB genes in B-like cells, respectively. Here, we identify and characterize several CDA1-like genes in the larvae of different lamprey species and demonstrate that these encode active cytidine deaminases. Structural comparisons of the CDA1 variants highlighted substantial differences in surface charge; this observation is supported by our finding that the enzymes require different conditions and substrates for optimal activity in vitro. Strikingly, we also found that the number of CDA-like genes present in individuals of the same species is variable. Nevertheless, irrespective of the number of different CDA1-like genes present, all lamprey larvae have at least one functional CDA1-related gene encoding an enzyme with predicted structural and chemical features generally comparable to jawed vertebrate AID. Our findings suggest that, similar to APOBEC3 branch expansion in jawed vertebrates, the AID/APOBEC family has undergone substantial diversification in lamprey, possibly indicative of multiple distinct biological roles.
Asunto(s)
Desaminasas APOBEC-1/genética , Citidina Desaminasa/clasificación , Citidina Desaminasa/genética , Variaciones en el Número de Copia de ADN , Lampreas/genética , Linfocitos/inmunología , Receptores de Antígenos/genética , Desaminasas APOBEC-1/química , Desaminasas APOBEC-1/inmunología , Secuencia de Aminoácidos , Animales , Citidina Desaminasa/química , Citidina Desaminasa/inmunología , Secuenciación de Nucleótidos de Alto Rendimiento , Conformación Proteica , Receptores de Antígenos/clasificación , Homología de Secuencia , Secuenciación Completa del GenomaRESUMEN
Among different bacteria colonizing the bovine uterus, Trueperella pyogenes is found to be associated with clinical endometritis (CE). The ability of cows to defend against T. pyogenes infections depends on the virulence of invading bacteria and on the host's innate immunity. Therefore, to gain insights into bacterial factors contributing to the interplay of this host pathogen, two strains of T. pyogenes were included in this study: one strain (TP2) was isolated from the uterus of a postpartum dairy cow developing CE and a second strain (TP5) was isolated from a uterus of a healthy cow. The two strains were compared in terms of their metabolic fingerprints, growth rate, virulence gene transcription, and effect on bovine endometrial epithelial cells in vitro. In addition, the effect of the presence of peripheral blood mononuclear cells (PBMCs) on the response of endometrial epithelial cells was evaluated. TP2, the strain isolated from the diseased cow, showed a higher growth rate, expressed more virulence factors (cbpA, nanH, fimE, and fimG), and elicited a higher mRNA expression of pro-inflammatory factors (PTGS2, CXCL3, and IL8) in bovine endometrial epithelial cells compared with TP5, the strain isolated from the healthy cow. The presence of PBMCs amplified the mRNA expression of pro-inflammatory factors (PTGS2, CXCL3, IL1A, IL6, and IL8) in bovine endometrial epithelial cells co-cultured with live TP2 compared with untreated cells, especially as early as after 4 h. In conclusion, particular strain characteristics of T. pyogenes were found to be important for the development of CE. Furthermore, immune cells attracted to the site of infection might also play an important role in up-regulation of the pro-inflammatory response in the bovine uterus and thus significantly contribute to the host-pathogen interaction.
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Actinomycetaceae/patogenicidad , Endometritis/veterinaria , Endometrio/inmunología , Endometrio/microbiología , Células Epiteliales/inmunología , Células Epiteliales/microbiología , Útero/inmunología , Útero/microbiología , Actinomycetaceae/genética , Actinomycetaceae/inmunología , Actinomycetaceae/aislamiento & purificación , Infecciones por Actinomycetales/inmunología , Infecciones por Actinomycetales/microbiología , Infecciones por Actinomycetales/veterinaria , Animales , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/microbiología , Técnicas de Cultivo de Célula , Supervivencia Celular , ADN Bacteriano , Endometritis/inmunología , Endometritis/microbiología , Femenino , Genes Bacterianos/genética , Interacciones Huésped-Patógeno/inmunología , Inmunidad Innata , Leucocitos Mononucleares , Periodo Posparto , ARN Mensajero/biosíntesis , Especificidad de la Especie , Regulación hacia Arriba , Virulencia/genética , Factores de Virulencia/genética , Factores de Virulencia/metabolismoRESUMEN
Endometrial epithelium plays a crucial role in the first immune response to invading bacteria by producing cytokines and chemokines. The aim of this study was to investigate the first inflammatory response of the endometrium in vivo and in vitro. Gene expression of several pro-inflammatory factors and Toll-like receptors (TLR2, -4, -6) was determined in endometrial cytobrush samples obtained from healthy cows and cows with clinical or subclinical endometritis. Endometrial epithelial cells were co-cultured with an isolated autochthonous uterine bacterial strain Bacillus pumilus. Total RNA was extracted from in vivo and in vitro samples and subjected to real-time reverse transcription polymerase chain reaction. CXC ligands (CXCL) 1/2 and CXC chemokine receptor (CXCR) 2 mRNA expression was higher in cows with subclinical endometritis and CXCL3 mRNA expression was higher in cows with clinical endometritis compared with healthy cows. B. pumilus induced cell death of epithelial cells within 24h of co-culturing. The presence of B. pumilus resulted in significantly higher mRNA expression of interleukin 1α (IL1A), IL6, IL8, CXCL1-3 and prostaglandin-endoperoxide synthase 2 in co-cultured cells compared with untreated controls. The maximum increase was mainly detected after 2h. These results support the hypothesis that bacterial infection of endometrial cells might induce prompt synthesis of pro-inflammatory cytokines resulting in a local inflammatory reaction.
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Bacillus pumilus , Bovinos , Endometrio/inmunología , Células Epiteliales/inmunología , ARN Mensajero/metabolismo , Animales , Células Cultivadas , Quimiocinas/inmunología , Citocinas/inmunología , Endometritis/inmunología , Endometrio/citología , Células Epiteliales/microbiología , Femenino , Receptores de Quimiocina/inmunologíaRESUMEN
Polycomb-mediated chromatin repression modulates gene expression during development in metazoans. Binding of multiple sequence-specific factors at discrete Polycomb response elements (PREs) is thought to recruit repressive complexes that spread across an extended chromatin domain. To dissect the structure of PREs, we applied high-resolution mapping of nonhistone chromatin proteins in native chromatin of Drosophila cells. Analysis of occupied sites reveal interactions between transcription factors that stabilize Polycomb anchoring to DNA, and implicate the general transcription factor ADF1 as a novel PRE component. By comparing two Drosophila cell lines with differential chromatin states, we provide evidence that repression is accomplished by enhanced Polycomb recruitment both to PREs and to target promoters of repressed genes. These results suggest that the stability of multifactor complexes at promoters and regulatory elements is a crucial aspect of developmentally regulated gene expression.
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Ensamble y Desensamble de Cromatina , Drosophila/genética , Proteínas del Grupo Polycomb/metabolismo , Elementos de Respuesta/genética , Animales , Células Cultivadas , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Unión Proteica , Factores de Transcripción/metabolismoRESUMEN
Although it is thought that changes in gene expression play an important role in adaptation, the identification of gene-regulatory sequences that have been targets of positive selection has proved difficult. Here, we identify a cis-regulatory element of the Drosophila melanogaster CG9509 gene that is associated with a selective sweep in a derived non-African population of the species. Expression analyses indicate that CG9509 consistently shows greater expression in non-African than in African strains of D. melanogaster. We find that a 1.8 kb region located just upstream of the CG9509 coding region is devoid of DNA sequence polymorphism in a European population sample and that this is best explained by the recent action of positive selection (within the past 4,000-10,000 years). Using a reporter gene construct and phiC31-mediated site-specific integration, we show that the European version of the CG9509 upstream region drives 2-3 times greater expression than the African version in an otherwise identical genetic background. This expression difference corresponds well to that of the native gene and indicates that sequence variation within the CG9509 upstream region can completely account for its high expression in the European population. Selection appears to have favored a quantitative increase in gene expression in the Malphigian tubule, the tissue where CG9509 is predominantly expressed.
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Drosophila melanogaster/genética , Genes de Insecto , Secuencias Reguladoras de Ácidos Nucleicos , Selección Genética/genética , Adaptación Biológica , Animales , Proteínas de Drosophila , Evolución Molecular , Femenino , Regulación de la Expresión Génica , Genética de Población , Masculino , Países Bajos , Polimorfismo Genético , ZimbabweRESUMEN
BACKGROUND: Variation at the level of gene expression is abundant in natural populations and is thought to contribute to the adaptive divergence of populations and species. Gene expression also differs considerably between males and females. Here we report a microarray analysis of gene expression variation among females of 16 Drosophila melanogaster strains derived from natural populations, including eight strains from the putative ancestral range in sub-Saharan Africa and eight strains from Europe. Gene expression variation among males of the same strains was reported previously. RESULTS: We detected relatively low levels of expression polymorphism within populations, but much higher expression divergence between populations. A total of 569 genes showed a significant expression difference between the African and European populations at a false discovery rate of 5%. Genes with significant over-expression in Europe included the insecticide resistance gene Cyp6g1, as well as genes involved in proteolysis and olfaction. Genes with functions in carbohydrate metabolism and vision were significantly over-expressed in the African population. There was little overlap between genes expressed differently between populations in females and males. CONCLUSIONS: Our results suggest that adaptive changes in gene expression have accompanied the out-of-Africa migration of D. melanogaster. Comparison of female and male expression data indicates that the vast majority of genes differing in expression between populations do so in only one sex and suggests that most regulatory adaptation has been sex-specific.
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Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Animales , Femenino , Perfilación de la Expresión Génica , Reacción en Cadena de la PolimerasaRESUMEN
The detection of selection, both positive and negative, acting on a DNA sequence or class of nucleotide sites requires comparison with a reference sequence that is unaffected by selection. In Drosophila, recent findings of widespread selective constraint, as well as adaptive evolution, in both coding and noncoding regions highlight the difficulties in choosing such a reference sequence. Here, we investigate the utility of short intron sequences as a reference for the detection of selection. For a set of 119 Drosophila melanogaster genes containing 195 short introns (Asunto(s)
Drosophila/genética
, Evolución Molecular
, Intrones
, Animales
, Bases de Datos Genéticas
, Ligamiento Genético
, Genética de Población
, Mutación INDEL
, Polimorfismo Genético
, Selección Genética
, Estadísticas no Paramétricas
RESUMEN
BACKGROUND: Bacterial and fungal infections induce a potent immune response in Drosophila melanogaster, but it is unclear whether viral infections induce an antiviral immune response. Using microarrays, we examined the changes in gene expression in Drosophila that occur in response to infection with the sigma virus, a negative-stranded RNA virus (Rhabdoviridae) that occurs in wild populations of D. melanogaster. PRINCIPAL FINDINGS: We detected many changes in gene expression in infected flies, but found no evidence for the activation of the Toll, IMD or Jak-STAT pathways, which control immune responses against bacteria and fungi. We identified a number of functional categories of genes, including serine proteases, ribosomal proteins and chorion proteins that were overrepresented among the differentially expressed genes. We also found that the sigma virus alters the expression of many more genes in males than in females. CONCLUSIONS: These data suggest that either Drosophila do not mount an immune response against the sigma virus, or that the immune response is not controlled by known immune pathways. If the latter is true, the genes that we identified as differentially expressed after infection are promising candidates for controlling the host's response to the sigma virus.
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Drosophila melanogaster/genética , Rhabdoviridae/patogenicidad , Animales , Secuencia de Bases , Cartilla de ADN , Regulación hacia Abajo , Drosophila melanogaster/inmunología , Drosophila melanogaster/virología , Femenino , Masculino , Reacción en Cadena de la Polimerasa , Regulación hacia ArribaRESUMEN
BACKGROUND: Differences in levels of gene expression among individuals are an important source of phenotypic variation within populations. Recent microarray studies have revealed that expression variation is abundant in many species, including Drosophila melanogaster. However, previous expression surveys in this species generally focused on a small number of laboratory strains established from derived populations. Thus, these studies were not ideal for population genetic analyses. RESULTS: We surveyed gene expression variation in adult males of 16 D. melanogaster strains from two natural populations, including an ancestral African population and a derived European population. Levels of expression polymorphism were nearly equal in the two populations, but a higher number of differences was detected when comparing strains between populations. Expression variation was greatest for genes associated with few molecular functions or biological processes, as well as those expressed predominantly in males. Our analysis also identified genes that differed in expression level between the European and African populations, which may be candidates for adaptive regulatory evolution. Genes involved in flight musculature and fatty acid metabolism were over-represented in the list of candidates. CONCLUSION: Overall, stabilizing selection appears to be the major force governing gene expression variation within populations. However, positive selection may be responsible for much of the between-population expression divergence. The nature of the genes identified to differ in expression between populations may reveal which traits were important for local adaptation to the European and African environments.