Anopheline salivary protein genes and gene families: an evolutionary overview after the whole genome sequence of sixteen Anopheles species

Background: Mosquito saliva is a complex cocktail whose pharmacological properties play an essential role in blood feeding by counteracting host physiological response to tissue injury. Moreover, vector borne pathogens are transmitted to vertebrates and exposed to their immune system in the context of mosquito saliva which, in virtue of its immunomodulatory properties, can modify the local environment at the feeding site and eventually affect pathogen transmission. In addition, the host antibody response to salivary proteins may be used to assess human exposure to mosquito vectors. Even though the role of quite a few mosquito salivary proteins has been clarified in the last decade, we still completely ignore the physiological role of many of them as well as the extent of their involvement in the complex interactions taking place between the mosquito vectors, the pathogens they transmit and the vertebrate host. The recent release of the genomes of 16 Anopheles species offered the opportunity to get insights into function and evolution of salivary protein families in anopheline mosquitoes. Results: Orthologues of fifty three Anopheles gambiae salivary proteins were retrieved and annotated from 18 additional anopheline species belonging to the three subgenera Cellia, Anopheles, and Nyssorhynchus. Our analysis included 824 full-length salivary proteins from 24 different families and allowed the identification of 79 novel salivary genes and re-annotation of 379 wrong predictions. The comparative, structural and phylogenetic analyses yielded an unprecedented view of the anopheline salivary repertoires and of their evolution over 100 million years of anopheline radiation shedding light on mechanisms and evolutionary forces that contributed shaping the anopheline sialomes. Conclusions: We provide here a comprehensive description, classification and evolutionary overview of the main anopheline salivary protein families and identify two novel candidate markers of human exposure to malaria vectors worldwide. This anopheline sialome catalogue, which is easily accessible as hyperlinked spreadsheet, is expected to be useful to the vector biology community and to improve the capacity to gain a deeper understanding of mosquito salivary proteins facilitating their possible exploitation for epidemiological and/or pathogen-vector-host interaction studies

Sudden cardiac death in young athletes: Literature review of molecular basis

Intense athletic training and competition can rarely result in sudden cardiac death (SCD). Despite the introduction of pre-participation cardiovascular screening, especially among young competitive athletes, sport-related SCD remains a debated issue among medical personnel, sports communities and laypersons alike, and generates significant media attention. The most frequent cause of SCD is a hidden inherited cardiomyopathy, the athletes may not even be aware of. Predictive medicine, by searching the presence of pathogenic alterations in cardiac genes, may be an integrative tool, besides the conventional ones used in cardiology (mainly electro and echocardiogram), to reach a definitive diagnosis in athletes showing signs/symptoms, even borderline, of inherited cardiomyopathy/ channelopathy, and in athletes presenting family history of SCD and/or of hereditary cardiac disease. In this review, we revised the molecular basis of the major cardiac diseases associated to sudden cardiac death and the clinical molecular biology approach that can be used to perform risk assessment at DNA level of sudden cardiac death, contributing to the early implementation of adequate therapy. Alterations can occur in ion channel genes, in genes encoding desmosomal and junctional proteins, sarcomeric and Z-disc proteins, proteins for the cytoskeleton and the nuclear envelope. The advent of next generation sequencing (NGS) technology has provided the means to search for mutations in all these genes, at the same time. Therefore, this molecular approach should be the preferred methodology for the aforementioned purpose

Antigeni salivari quali strumenti epidemiologici per la valutazione dell'esposizione umana ad Aedes albopictus

Hematophagous arthropods during feeding inject into their hosts a cocktail of salivary proteins whose main role is to allow for an effective blood meal by counteracting host hemostasis, inflammation and immunity. However, saliva of blood feeders also evokes in vertebrates an antibody response that can be used to evaluate exposure to disease vectors. Salivary transcriptome studies carried out in different hematophagous species in the last fifteen years clarified the complexity of the salivary repertoires of blood feeding arthropods, pointing out that salivary proteins evolve at a fast evolutionary rate and highlighting the existence of family-, genus- and sometime even species-specific salivary proteins. Focusing on mosquitoes of the genera Anopheles and Aedes, which are important vectors of the human malaria parasite Plasmodium falciparum and of several arboviruses, we summarize here recent efforts to exploit genus-specific salivary proteins as biomarkers of human exposure to these vectors of large relevance for public health

MicroRNAs from saliva of anopheline mosquitoes mimic human endogenous miRNAs and may contribute to vector-host-pathogen interactions

During blood feeding haematophagous arthropods inject into their hosts a cocktail of salivary proteins whose main role is to counteract host haemostasis, inflammation and immunity. However, animal body fluids are known to also carry miRNAs. To get insights into saliva and salivary gland miRNA repertoires of the African malaria vector Anopheles coluzzii we used small RNA-Seq and identified 214 miRNAs, including tissue-enriched, sex-biased and putative novel anopheline miRNAs. Noteworthy, miRNAs were asymmetrically distributed between saliva and salivary glands, suggesting that selected miRNAs may be preferentially directed toward mosquito saliva. The evolutionary conservation of a subset of saliva miRNAs in Anopheles and Aedes mosquitoes, and in the tick Ixodes ricinus, supports the idea of a non-random occurrence pointing to their possible physiological role in blood feeding by arthropods. Strikingly, eleven of the most abundant An. coluzzi saliva miRNAs mimicked human miRNAs. Prediction analysis and search for experimentally validated targets indicated that miRNAs from An. coluzzii saliva may act on host mRNAs involved in immune and inflammatory responses. Overall, this study raises the intriguing hypothesis that miRNAs injected into vertebrates with vector saliva may contribute to host manipulation with possible implication for vector-host interaction and pathogen transmission

Fine-scale differences in diel activity among nocturnal freshwater planarias (Platyhelminthes: Tricladida)

<p>Abstract</p> <p>Background</p> <p>Although most freshwater planarias are well known photonegative organisms, their diel rhythms have never been quantified. Differences in daily activity rhythms may be particularly important for temperate-climate, freshwater planarias, which tend to overlap considerably in spatial distribution and trophic requirements.</p> <p>Methods</p> <p>Activity of stress-free, individually tested young adults of three common planarian species was recorded at 3-h intervals in a 10-d experiment under natural sunlight and photoperiod during autumnal equinox (D:L ~12:12). Individual activity status was averaged over the 10-d experiment, each tested individual thus serving as a true replicate. Twelve individuals per species were tested. Food was provided every 36 h, resulting in alternating day- and nighttime feeding events. Activity during the first post-feeding h was recorded and analyzed separately. Statistical procedures included ANOVAs, correlations, and second-order analyses of angles.</p> <p>Results</p> <p><it>Dugesia (= Girardia) tigrina </it>Girard 1850 exhibited clear nocturnal behavior, <it>Dugesia (= Schmidtea) polychroa </it>Schmidt 1861 was predominantly but not exclusively nocturnal, and <it>Polycelis tenuis </it>Ijima 1884 was relatively more active from midnight through noon. Species-specific activity peaks were statistically similar, with peaks at dawn for <it>P. tenuis </it>and just before midnight for the two dugesiids; however, <it>D. tigrina </it>was comparatively more active in the early night hours, while <it>D. polychroa </it>was more active than <it>D. tigrina </it>during daytime. <it>D. tigrina </it>also responded less readily to daytime food addition. <it>P. tenuis </it>remained poorly active and unresponsive throughout the experiment. Individual variability in diel behavior was highest for <it>D. polychroa </it>and lowest for <it>D. tigrina. P. tenuis</it>'s general low degree of activity and late activity peak in the experiment may be related to a strong reliance on external stimuli.</p> <p>Conclusions</p> <p>The tested species are mainly nocturnal, consistent with their photonegative characteristics. The fine-scale differences in diel behavior among these three triclad species may not be sufficient to allow coexistence in the wild, with the nonnative <it>D. tigrina </it>eventually displacing <it>D. polychroa </it>and <it>P. tenuis </it>in many European waters. The link between planarian diel rhythms and ecological characteristics are worth of further, detailed investigation.</p

Recital de grado consideraciones técnicas para un recital de flauta

El objetivo de este trabajo es brindar información concreta sobre diversos temas concernientes a la flauta, comenzado por la importancia de elegir un instrumento y que factores intervienen al momento de tomar esta decisión. Por otro lado, el documento expone las obras que fueron interpretadas durante el recital. En este aspecto, se investigó sobre la vida de sus compositores a fin de conocer su contexto para una ejecución históricamente informada. Además, se hizo un análisis musical que abarca aspectos como la rítmica, las formas musicales, las escalas etc. con la finalidad de nutrir el conocimiento de estas. Por último, se encuentra una serie de recomendaciones sobre dichas obras con el objetivo de aportar las experiencias conseguidas a través de esta investigación para que el lector consiga una orientación (qué tipo de orientación) de dichos temas

Analysis of apyrase 5' upstream region validates improved Anopheles gambiae transformation technique

<p>Abstract</p> <p>Background</p> <p>Genetic transformation of the malaria mosquito <it>Anopheles gambiae </it>has been successfully achieved in recent years, and represents a potentially powerful tool for researchers. Tissue-, stage- and sex-specific promoters are essential requirements to support the development of new applications for the transformation technique and potential malaria control strategies. During the <it>Plasmodium </it>lifecycle in the invertebrate host, four major mosquito cell types are involved in interactions with the parasite: hemocytes and fat body cells, which provide humoral and cellular components of the innate immune response, midgut and salivary glands representing the epithelial barriers traversed by the parasite during its lifecycle in the mosquito.</p> <p>Findings</p> <p>We have analyzed the upstream regulatory sequence of the <it>An. gambiae </it>salivary gland-specific <it>apyrase </it>(<it>AgApy</it>) gene in transgenic <it>An. gambiae </it>using a <it>piggyBac </it>transposable element vector marked by a <it>3xP3 </it>promoter:<it>DsRed </it>gene fusion. Efficient germ-line transformation in <it>An. gambiae </it>mosquitoes was obtained and several integration events in at least three different G₀families were detected. <it>LacZ </it>reporter gene expression was analyzed in three transgenic lines/groups, and in only one group was tissue-specific expression restricted to salivary glands.</p> <p>Conclusion</p> <p>Our data describe an efficient genetic transformation of <it>An. gambiae </it>embryos. However, expression from the selected region of the <it>AgApy </it>promoter is weak and position effects may mask tissue- and stage- specific activity in transgenic mosquitoes.</p