Ocorrência de Escherichia coli 0157:H7 em vegetais e resistência aos agentes de desinfecção de verduras

Foi feito um estudo da ocorrência de E.coli O157:H7 em vegetais que são normalmente consumidos crus no Brasil e uma avaliação da sua resistência aos sanitizantes disponíveis no mercado para desinfecção de verduras, equipamentos e utensílios, incluindo compostos clorados e compostos de amônio quaternário. Na avaliação da ocorrência em vegetais foram analisadas 869 amostras, não sendo detectada a presença do patógeno. Os imunoensaios enzimáticos (ELISA) utilizados nas análises (Reveal E.coli O157 Neogem e EHEC Test Kit 3M Company) apresentaram uma taxa de falsos resultados presuntivos de 13,6 e 11,8%, respectivamente, não confirmados como E.coli O157 nos testes bioquímicos posteriores. Na avaliação da resistência aos sanitizantes pelo método 960.9 da AOAC, observou-se que os tratamentos com 100 e 200ppm de hipoclorito de sódio, dicloroisocianurato de sódio e cloreto de benzalcônio/30s se mostraram eficazes contra E.coli ATCC 11229 e E.coli O157:H7 ATCC 43890, promovendo mais de 5 reduções decimais nas populações alvo.

Prevalência, quantificação e resistência a antimicrobianos de sorovares de Salmonella isolados de lingüiça frescal suína

A Salmonella sp. é uma das principais causas mundiais de toxinfecção alimentar. Nos últimos anos, as preocupações têm se voltado para a carne e produtos suínos tanto no aspecto de saúde pública como na sua comercialização/exportação. O presente estudo tem como objetivos: 1) verificar a prevalência de sorovares de Salmonella sp. em lingüiças tipo frescal de matéria-prima suína comercializadas em Lages (SC), bem como o seu nível de contaminação; e 2) verificar o perfil de resistência aos antimicrobianos destes isolados. Para tanto, foram coletadas 200 amostras de nove marcas, em diferentes estabelecimentos comerciais. Foram isoladas Salmonella sp. em 27% (54), sendo o sorovar Derby o mais encontrado. Apenas uma amostra apresentou uma concentração de microorganismos maior que 1,100 NMP.g-1, valor normalmente tido como necessário para causar infecção por Salmonella do grupo não-tifóide. Posteriormente, os 60 isolados foram submetidos ao teste de susceptibilidade in vitro, frente a 14 antimicrobianos. Entre esses isolados, 56,67% apresentaram resistência a pelo menos um dos antimicrobianos testados e o perfil de multirresistência foi encontrado em 20%. A prevalência elevada de produtos positivos para Salmonella sp. pode representar um risco ao consumidor, principalmente considerando-se o alto número de isolados resistentes encontrado neste estudo.

Resistência antimicrobiana de Pseudomonas aeruginosa isolados de pescado e de cortes e de miúdos de frango

Pseudomonas aeruginosa isolados de peixes de água doce e de frangos foram submetidos ao teste de suscetibilidade aos antimicrobianos utilizando quatorze drogas, com o objetivo de determinar e confrontar os padrões de suscetibilidade deste microrganismo. As cepas oriundas de peixes pertenciam à coleção do Laboratório de Bacterioses/IV/UFRRJ. Para o isolamento das cepas, foram selecionados miúdos (fígado) e cortes (coxa e sobrecoxa) de frangos adquiridos em estabelecimentos comerciais no município do Rio de Janeiro. A metodologia de isolamento incluiu o enriquecimento em água peptonada, seguido de semeadura em Agar EMB e Agar GSP. Para as cepas oriundas de peixes, procedeu-se à reativação em água peptonada, seguida de reisolamento em Agar EMB. Colônias sugestivas foram transferidas para Agar TSI e LIA para avaliação das características metabólicas. A capacidade de produção de pigmento verde-azulado foi avaliada em Agar Mueller-Hinton e a da enzima citocromo-oxidase, em Agar Nutriente. O teste de suscetibilidade a antimicrobianos realizado nas 63 cepas revelou maiores percentuais de resistência para NAL e NIT (96,8%), TCY (93,6%), AMC (92,1%), CHL (90,5%) e SXT (85,7%), destacando-se a multirresistência dos isolados. A totalidade das cepas oriundas de frangos apresentou sensibilidade a CAZ e IPM e nos isolados de peixes a ATM, CAZ, IPM e AMK.

Qualidade fisiológica e resistência do recobrimento de sementes de milho

Este trabalho foi desenvolvido com o objetivo de avaliar a qualidade fisiológica de sementes de milho UENF 506-8 recobertas e a resistência do recobrimento utilizando diferentes proporções de cimentante. Foi utilizada no recobrimento das sementes uma mistura de calcário e meio de cultura. Como cimentante foi utilizada cola a base de acetato de polivinila (PVA). O recobrimento das sementes de milho foi realizado com uma mistura de 70% (p/p) de calcário, 10% (p/p) de meio de cultura semi-sólido JNFb e 20% (p/p) de água. Foram adicionadas a essa mistura diferentes proporções de cimentante: 0,5; 1; 2; 3; 4 e 5 % (p/p). Para avaliação da resistência do recobrimento foi simulado o transporte e a semeadura manual, e a semeadura mecânica das sementes recobertas. A avaliação da qualidade fisiológica das sementes foi realizada em sementes nuas (controle) e sementes recobertas com as proporções de cimentante: 0,5; 1; 2; 3; 4 e 5 %. O aumento da proporção de cimentante de 2 para 3% causou uma pequena redução na velocidade da germinação, na velocidade de emergência e na emergência, mas não afetou o número final de plântulas normais no teste de germinação e teste frio. Para a semeadura manual recomenda-se a utilização de 2% de cimentante e para a semeadura mecanizada 4 % de cimentante.

Qualidade fisiológica de sementes de soja tratadas com inseticidas sob efeito do armazenamento

Dentre os conceitos modernos de controle de pragas, o uso de inseticidas no tratamento de sementes constitui-se num dos métodos mais eficientes. Deve-se, entretanto, conhecer a influência desses produtos com relação à qualidade fisiológica das sementes tratadas. Objetivou-se através desta pesquisa avaliar a qualidade fisiológica de sementes de soja tratadas com inseticidas, sob quatro períodos de armazenamento (0, 15, 30 e 45 dias após o tratamento). O delineamento experimental foi o inteiramente casualizado, em esquema fatorial 4 x 7, com quatro repetições, onde as sementes do cultivar M-SOY 6101 foram tratadas com os inseticidas thiamethoxam (Cruiser 700 WS), na dose de 250 mL de produto comercial (p.c.)/100 kg de sementes; fipronil (Standak), na dose de 150 mL de p.c./100 kg de sementes; imidacloprid (Gaucho FS), na dose de 150 ml de p.c./100 kg de sementes; [imidacloprid + thiodicarb] (CropStar FS), na dose de 0,3 L. ha-1; carbofuran (Furadan 350 TS), na dose de 1,5 L/100 kg de sementes; acefato (Orthene 750 BR), na dose de 1 kg/ 100 kg de sementes e uma testemunha, sem tratamento. As variáveis analisadas foram: germinação, velocidade de emergência, comprimento de raiz e de plântula e porcentagem de plântulas normais no teste de envelhecimento acelerado. A aplicação dos inseticidas carbofuran e acefato é prejudicial à qualidade fisiológica das sementes de soja cultivar M-SOY 6101, por um período de armazenamento de até 45 dias. A redução da qualidade fisiológica das sementes, condicionada pelos inseticidas avaliados, intensifica-se com o prolongamento do armazenamento das sementes tratadas, recomendando-se, portanto, que o tratamento inseticida das sementes seja realizado próximo ao momento da semeadura.

Tratamento de sementes de sorgo com inseticidas

O tratamento de sementes com inseticidas é um método eficiente que auxilia o manejo integrado de pragas, porém, há escassez de informações de produtos que apresentem seletividade para a cultura do sorgo. O objetivo deste trabalho foi o de avaliar a influência do tratamento de sementes de sorgo com inseticidas, contendo um ou dois princípios ativos, sem e com o armazenamento por 30 dias, na fase inicial de desenvolvimento das plantas. O experimento foi conduzido na Fesurv-Universidade de Rio Verde, em Rio Verde-GO com dez inseticidas (thiodicarb, fipronil, acephate, thiametoxam, imidacloprid, [imidacloprid+thiodicarb], imidacloprid+thiodicarb, imidacloprid+fipronil, thiametoxam+thiodicarb, thiametoxam+fipronil, acrescido da testemunha sem inseticida) sem e com o armazenamento das sementes por 30 dias. Os resultados obtidos permitiram constatar que o armazenamento por 30 dias das sementes de sorgo tratadas com inseticidas diminui o percentual de germinação. A mistura imidaclopridrid+thiodicarb apresenta maior potencial para uso no tratamento de sementes de sorgo. O inseticida acephate causa fitotoxicidade às plântulas de sorgo ao contrário do fipronil, thiametoxam e a mistura thiametoxam+thiodicarb que não prejudicam o vigor das sementes de sorgo.

Analysis of a temperature sensitive mutation affecting aldehyde oxidase activity in Drosophila melanogaster

A strain of Drosophila melanogaster (mid america stock culture no. hl16) has been reported to be deficient in aldehyde oxidase activity (Hickey and Singh 1982). This strain was characterized during the course of this study and compared to other mutant strains known to be deficient in aldehyde oxidase activity. During the course of this investigation, the hl16 strain was found to be temperature sensitive in its viability. It was found that the two phenotypes, the enzyme deficiency, and the temperature sensitive lethality were the result of two different mutations, both mapping to the X-chromosome. These two mutations were found to be separable by recombination. The enzyme deficiency was found to map to the same locus as the cinnamon mutation, another mutation which affects aldehyde oxidase production. The developmental profile of aldehyde oxidase in the hl16 strain was compared to the developmental profile in the Canton S wild type strain. The aldehyde oxidase activity in adult hl16 individuals was also compared to that of various other strains. It was also found that the aldehyde oxidase activity was temperature sensitive in the adult flies. The temperature sensitive lethality mutation was mapped to position 1-0.1.

Viability interactions between the left and right arms of the second chromosome of Drosophila melanogaster

Inter and intrachromosomal viability interactions have been detected in a few experimental studies. Computer simulations and analytical models have led to postulation of nonadditivity of gene action. This study reports evidence of strong nonadditive interactions between the arms of the metacentric second chromosome of Drosophila melanogaster. Mean viability for 40 homozygous lines of the second chromosomes was 0.720+0.265 • Mean viability for 40 half homozygous second chromosomes was 0.928!O.)10 • Significant heterogeneity among and within lines was found in both groups of chromosomes, as well as a highly significant viability difference between the two groups. Comparison of observed viabilities with the expected values, according to the theories of additive and multi - plicative gene action. was made for both groups. Highly significant departures from the expected values were found for over 90% of the lines in both groups of chromosomes, for both additive and multiplicative models of gene action.

Frequency-dependent selection at the amylase locus in Drosophila melanogaster

A. strain of Drosophila melanog-aster deficient in null amylase activity (Amylase ) was isolated from a wild null population of flies. The survivorship of Amylase homozygous flies is very low when the principal dietary carbohydrate source is starch. However, the survivorship of the null Amylase genotype is comparable to the wild type when the dietary starch is replaced by glucose. In addition, the null viability of the amylase-producing and Amylase strains is comparable v and very lm<] f on a medium with no carbohydrates . Furthermore, amylase-producing genotypes were shovm to excrete enzymatically active amylase protein into the food medium. The excreted amylase causes the external breakdown of dietary starch to sugar. These results led to the following null prediction: the viability of the A.mvlase genotype (fed on a starch rich diet) might increase in the presence of individuals which were amylase-producing. It was shown experimentally that such an increase in viability did in fact occur and that this increase v\Tas proportional to the number of mnylase..::producing fli.es present. These results provide a unique example of a non-"competi ti ve inter-genotype interaction, and one where the underlying physio~ logical and biochemical mechanism has been fully understood.

Action of octopamine and tyramine on muscles of Drosophila melanogaster larvae

Octopamine (OA) and tyramine (TA) play important roles in homeostatic mechanisms, behavior, and modulation of neuromuscular junctions in arthropods. However, direct actions of these amines on muscle force production that are distinct from effects at the neuromuscular synapse have not been well studied. We utilize the technical benefits of the Drosophila larval preparation to distinguish the effects of OA and TA on the neuromuscular synapse from their effects on contractility of muscle cells. In contrast to the slight and often insignificant effects of TA, the action of OA was profound across all metrics assessed. We demonstrate that exogenous OA application decreases the input resistance of larval muscle fibers, increases the amplitude of excitatory junction potentials (EJPs), augments contraction force and duration, and at higher concentrations (10−5 and 10−4 M) affects muscle cells 12 and 13 more than muscle cells 6 and 7. Similarly, OA increases the force of synaptically driven contractions in a cell-specific manner. Moreover, such augmentation of contractile force persisted during direct muscle depolarization concurrent with synaptic block. OA elicited an even more profound effect on basal tonus. Application of 10−5 M OA increased synaptically driven contractions by ∼1.1 mN but gave rise to a 28-mN increase in basal tonus in the absence of synaptic activation. Augmentation of basal tonus exceeded any physiological stimulation paradigm and can potentially be explained by changes in intramuscular protein mechanics. Thus we provide evidence for independent but complementary effects of OA on chemical synapses and muscle contractility.

The action of octopamine on muscles of Drosophila melanogaster larvae

Octopamine (OA) and tyramine (TA) play important roles in homeostatic mechanisms, behavior, and modulation of neuromuscular junctions in arthropods. However, direct actions of these amines on muscle force production that are distinct from effects at the neuromuscular synapse have not been well studied. We utilize the technical benefits of the Drosophila larval preparation to distinguish the effects of OA and TA on the neuromuscular synapse from their effects on contractility of muscle cells. In contrast to the slight and often insignificant effects of TA, the action of OA was profound across all metrics assessed. We demonstrate that exogenous OA application decreases the input resistance of larval muscle fibers, increases the amplitude of excitatory junction potentials (EJPs), augments contraction force and duration, and at higher concentrations (10(-5) and 10(-4) M) affects muscle cells 12 and 13 more than muscle cells 6 and 7. Similarly, OA increases the force of synaptically driven contractions in a cell-specific manner. Moreover, such augmentation of contractile force persisted during direct muscle depolarization concurrent with synaptic block. OA elicited an even more profound effect on basal tonus. Application of 10(-5) M OA increased synaptically driven contractions by ≈ 1.1 mN but gave rise to a 28-mN increase in basal tonus in the absence of synaptic activation. Augmentation of basal tonus exceeded any physiological stimulation paradigm and can potentially be explained by changes in intramuscular protein mechanics. Thus we provide evidence for independent but complementary effects of OA on chemical synapses and muscle contractility.

Characterization of the mammalian homologs of the Drosophila Melanogaster Endocytic protein lethal (2) giant discs 1

Endocytose joue un rôle dans l'activation du récepteur Notch. Des mutations dans le gène drosophilien lethal giant discs (lgd), provoque une prolifération cellulaire en perturbant l'endocytose de Notch. Les orthologues murins mlgd1 et 2 peuvent sauver ce phénotype, démontrant une fonction conservée. Cependant, des publications récentes suggèrent que les orthologs humains de lgd (hgd1/2) sont nucléaires. Dans cette étude, il est démontré que chez la Drosophile, le mutant dlgd(08) provoque l'accumulation de Notch dans des vésicules et une surprolifération de neuroblastes . Ceci suggère que Notch est activé a l'intérieur des endosomes dans les neuroblastes. L'immunohistochimie de cellules Hela indique que hlgd1 et 2 ne sont pas nucléaires, mais associés à des strctures endosomales. Enfin, la baisse d'expression par shRNA des gènes murins mlgd1 et mlgd2 provoque une différenciation accélérée des cellules souches hématopoïétiques dans la lignée lymphopoïèse T et bloque la transition DN3 / CD4+CD8+, suggérant une suractivation de Notch.

Functional genomics of the Drosophila melanogaster X chromosome and the role of DWnt5 during development

The collection of X chromosome insertions (PX) lethal lines, which was isolated from a screen for essential genes on the X chromosome, was characterized by means of cloning the insertion sites, mapping the sites within genomic DNA and determination of the associated reporter gene expresssion patterns. The established STS flanking the P element insertion sites were submitted to EMBL nucleotide databases and their in situ data together with the enhancer trap expression patterns have been deposited in the FlyView database. The characterized lines are now available to be used by the scientific community for a detailed analysis of the newly established lethal gene functions. One of the isolated genes on the X chromosome was the Drosophila gene Wnt5 (DWnt5). From two independent screens, one lethal and three homozygous viable alleles were recovered, allowing the identification of two distinct functions for DWnt5 in the fly. Observations on the developing nervous system of mutant embryos suggest that DWnt5 activity affects axon projection pattern. Elevated levels of DWNT5 activity in the midline cells of the central nervous system causes improper establishment and maintenance of the axonal pathways. Our analysis of the expression and mutant phenotype indicates that DWnt5 function in a process needed for proper organization of the nervous system. A second and novel function of DWnt5 is the control of the body size by regulation of the cell number rather than affecting the size of cells. Moreover, experimentally increased DWnt5 levels in a post-mitotic region of the eye imaginal disc causes abnormal cell cycle progression, resulting in additional ommatidia in the adult eye when compared to wild type. The increased cell number and the effects on the cell cycle after exposure to high DWNT5 levels is the result of a failure to downregulate cyclin B and therefore the unsuccessful establishment of a G1 arrest.

The role of Ken&Barbie in JAK/STAT signalling during development of Drosophila melanogaster

Cell-cell interactions during embryonic development are crucial in the co-ordination of growth, differentiation and maintenance of many different cell types. To achieve this co-ordination each cell must properly translate signals received from neighbouring cells, into spatially and temporally appropriate developmental responses. A surprisingly limited number of signal pathways are responsible for the differentiation of enormous variety of cell types. As a result, pathways are frequently 'reused' during development. Thus, in mammals the JAK/STAT pathway is required during early embryogenesis, mammary gland formation, hematopoiesis and, finally, plays a pivotal role in immune response. In the canonical way, the JAK/STAT pathway is represented by a transmembrane receptor associated with a Janus kinase (JAK), which upon stimulation by an extra-cellular ligand, phosphorylates itself, the receptor and, finally, the signal transducer and activator of transcription (STAT) molecules. Phosphorylated STATs dimerise and translocate to the nucleus where they activate transcription of target genes. The JAK/STAT pathway has been conserved throughout evolution, and all known components are present in the genome of Drosophila melanogaster. Besides hematopoietic and immunity functions, the pathway is also required during development for processes including embryonic segmentation, tracheal morphogenesis, posterior spiracle formation etc. This study describes Drosophila Ken&Barbie (Ken) as a selective regulator of JAK/STAT signalling. ken mutations identified in a screen for modulators of an eye overgrowth phenotype, caused by over-expression of the pathway ligand unpaired, also interact genetically with the pathway receptor domeless (dome) and the transcription factor stat92E. Over-expression of Ken can phenocopy developmental defects known to be caused by the loss of JAK/STAT signalling. These genetic interactions suggest that Ken may function as a negative regulator of the pathway. Ken has C-terminal Zn-finger domain, presumably for DNA binding, and N-terminal BTB/POZ domain, often found in transcriptional repressors. Using EGFP-fused construct expressed in vivo revealed nuclear accumulation of Ken. Therefore, it is proposed that Ken may act as a suppresser of STAT92E target genes. An in vitro assay, termed SELEX, determined that Ken specifically binds to a DNA sequence, with the essential for DNA recognition core overlapping that of STAT92E. This interesting observation suggests that not all STAT92E sites may also allow Ken binding. Strikingly, when effects of ectopic Ken on the expression of putative JAK/STAT pathway target genes were examined, only a subset of the genes tested, namely vvl, trh and kni, were down-regulated by Ken, whereas some others, such as eve and fj, appeared to be unresponsive. Further analysis of vvl, one of the genes susceptible to ectopic Ken, was undertaken. In the developing hindgut, expression of vvl is JAK/STAT pathway dependent, but remains repressed in the posterior spiracles, despite the stimulation of STAT92E by Upd in their primordia. Importantly, ken is also expressed in the developing posterior spiracles. Strikingly, up-regulation of vvl is observed in these tissues in ken mutant embryos. These imply that while ectopic Ken is sufficient to repress the expression of vvl in the hindgut, endogenous Ken is also necessary to prevent its activation in the posterior spiracles. It is therefore conceivable that ectopic vvl expression in the posterior spiracles of the ken mutants may be the result of de-repression of endogenous STAT92E activity. Another consequence of these observations is a fine balance that must exist between STAT92E and Ken activities. Apparently, endogenous level of Ken is sufficient to repress vvl, but not other, as yet unidentified, JAK/STAT pathway targets, whose presumable activation by STAT92E is required for posterior spiracle development as the embryos mutant for dome, the receptor of the pathway, show severe spiracle defects. These defects are also observed in the embryos mis-expressing Ken. Though it is possible that the posterior spiracle phenotype caused by higher levels of Ken results from a JAK/STAT pathway independent activity, it seems to be more likely that Ken acts in a dosage dependent manner, and extra Ken is able to further antagonise JAK/STAT pathway target genes. While STAT92E binding sites required for target gene expression have been poorly characterised, the existence of genome data allows the prediction of candidate STAT92E sites present in target genes promoters to be attempted. When a 6kb region containing the putative regulatory domains flanking the vvl locus are examined, only a single potential STAT92E binding site located 825bp upstream of the translational start can be detected. Strikingly, this site also includes a perfect Ken binding sequence. Such an in silico observation, though consistent with both Ken DNA binding assay in vitro and regulation of STAT92E target genes in vivo, however, requires further analysis. The JAK/STAT pathway is implicated in a variety of processes during embryonic and larval development as well as in imago. In each case, stimulation of the same transcription factor results in different developmental outcomes. While many potential mechanisms have been proposed and demonstrated to explain such pleiotropy, the present study indicates that Ken may represent another mechanism, with which signal transduction pathways are controlled. Ken selectively down-regulates a subset of potential target genes and so modifies the transcriptional profile generated by activated STAT92E - a mechanism, which may be partially responsible for differences in the morphogenetic processes elicited by JAK/STAT signalling during development.

Studies of cap-independent mRNA translation in Drosophila melanogaster

Control of protein synthesis is a key step in the regulation of gene expression during apoptosis and the heat shock response. Under such conditions, cap-dependent translation is impaired and Internal Ribosome Entry Site (IRES)-dependent translation plays a major role in mammalian cells. Although the role of IRES-dependent translation during apoptosis has been mainly studied in mammals, its role in the translation of Drosophila apoptotic genes has not been yet studied. The observation that the Drosophila mutant embryos for the cap-binding protein, the eukaryotic initiation factor eIF4E, exhibits increased apoptosis in correlation with up-regulated proapoptotic gene reaper (rpr) transcription constitutes the first evidence for the existence of a cap-independent mechanism for the translation of Drosophila proapoptotic genes. The mechanism of translation of rpr and other proapoptotic genes was investigated in this work. We found that the 5 UTR of rpr mRNA drives translation in an IRES-dependent manner. It promotes the translation of reporter RNAs in vitro either in the absence of cap, in the presence of cap competitors, or in extracts derived from heat shocked and eIF4E mutant embryos and in vivo in cells transfected with reporters bearing a non functional cap structure, indicating that cap recognition is not required in rpr mRNA for translation. We also show that rpr mRNA 5 UTR exhibits a high degree of similarity with that of Drosophila heat shock protein 70 mRNA (hsp70), an antagonist of apoptosis, and that both are able to conduct IRES-mediated translation. The proapoptotic genes head involution defective (hid) and grim, but not sickle, also display IRES activity. Studies of mRNA association to polysomes in embryos indicate that both rpr, hsp70, hid and grim endogenous mRNAs are recruited to polysomes in embryos in which apoptosis or thermal stress was induced. We conclude that hsp70 and, on the other hand, rpr, hid and grim which are antagonizing factors during apoptosis, use a similar mechanism for protein synthesis. The outcome for the cell would thus depend on which protein is translated under a given stress condition. Factors involved in the differential translation driven by these IRES could play an important role. For this purpose, we undertook the identification of the ribonucleoprotein (RNP) complexes assembled onto the 5 UTR of rpr mRNA. We established a tobramycin-affinity-selection protocol that allows the purification of specific RNP that can be further analyzed by mass spectrometry. Several RNA binding proteins were identified as part of the rpr 5 UTR RNP complex, some of which have been related to IRES activity. The involvement of one of them, the La antigen, in the translation of rpr mRNA, was established by RNA-crosslinking experiments using recombinant protein and rpr 5 UTR and by the analysis of the translation efficiency of reporter mRNAs in Drosophila cells after knock down of the endogenous La by RNAi experiments. Several uncharacterized proteins were also identified, suggesting that they might play a role during translation, during the assembly of the translational machinery or in the priming of the mRNA before ribosome recognition. Our data provide evidence for the involvement of La antigen in the translation of rpr mRNA and set a protocol for purification of tagged-RNA-protein complexes from cytoplasmic extracts. To further understand the mechanisms of translation initiation in Drosophila, we analyzed the role of eIF4B on cap-dependent and cap-independent translation. We showed that eIF4B is mostly involved in cap-, but not IRES-dependent translation as it happens in mammals.