Silencing of genes involved in Anaplasma marginale-tick interactions affects the pathogen developmental cycle in Dermacentor variabilis

<p>Abstract</p> <p>Background</p> <p>The cattle pathogen, <it>Anaplasma marginale</it>, undergoes a developmental cycle in ticks that begins in gut cells. Transmission to cattle occurs from salivary glands during a second tick feeding. At each site of development two forms of <it>A. marginale </it>(reticulated and dense) occur within a parasitophorous vacuole in the host cell cytoplasm. However, the role of tick genes in pathogen development is unknown. Four genes, found in previous studies to be differentially expressed in <it>Dermacentor variabilis </it>ticks in response to infection with <it>A. marginale</it>, were silenced by RNA interference (RNAi) to determine the effect of silencing on the <it>A. marginale </it>developmental cycle. These four genes encoded for putative glutathione S-transferase (GST), salivary selenoprotein M (SelM), H+ transporting lysosomal vacuolar proton pump (vATPase) and subolesin.</p> <p>Results</p> <p>The impact of gene knockdown on <it>A. marginale </it>tick infections, both after acquiring infection and after a second transmission feeding, was determined and studied by light microscopy. Silencing of these genes had a different impact on <it>A. marginale </it>development in different tick tissues by affecting infection levels, the densities of colonies containing reticulated or dense forms and tissue morphology. Salivary gland infections were not seen in any of the gene-silenced ticks, raising the question of whether these ticks were able to transmit the pathogen.</p> <p>Conclusion</p> <p>The results of this RNAi and light microscopic analyses of tick tissues infected with <it>A. marginale </it>after the silencing of genes functionally important for pathogen development suggest a role for these molecules during pathogen life cycle in ticks.</p

Caracterização morfobiológica, morfométrica e ultraestrutural de isolados silvestres de Trypanosoma cruzi do estado do Rio de Janeiro, Brasil

Triatoma vitticeps is a triatomine with geographic distribution restrict to Brazil, which exhibits high prevalence of Trypanosoma cruzi natural infection. Of special epidemiologic concern, this species often invades households in the states of Rio de Janeiro, Minas Gerais and Espírito Santo. The objective of this study was to evaluate morphological and ultrastructural parameters on three T. cruzi isolates obtained from wild T. vitticeps specimens. The growth and cell differentiation of the parasite was evaluated through epimastigote and trypomastigote forms obtained in the growth curves for three distinct isolates. The maximum growth showed differences at the 20th day of the curve. Our in vitro results show a heterogeneity, regarding these features for samples cultivated under the same conditions. Morphometric analyzes based on the shape of epimastigotes and trypomastigotes corroborated such differentiation. These results highlight the need of better understanding the meaning of this diversity under an eco-epidemiological perspective.Fil: Da Silva, C. Santos. Instituto Oswaldo Cruz; Brasil. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Carbajal de la Fuente, Ana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Almeida, C.E.. Universidade Estadual de Campinas; BrasilFil: Gonçalves, T.C.M.. Instituto Oswaldo Cruz; BrasilFil: Dos Santos Mallet, J. Reis. Instituto Oswaldo Cruz; Brasi

Caracterização morfobiológica, morfométrica e ultraestrutural de isolados silvestres de Trypanosoma cruzi do estado do Rio de Janeiro, Brasil

Triatoma vitticeps is a triatomine with geographic distribution restrict to Brazil, which exhibits high prevalence of Trypanosoma cruzi natural infection. Of special epidemiologic concern, this species often invades households in the states of Rio de Janeiro, Minas Gerais and Espirito Santo. The objective of this study was to evaluate morphological and ultrastructural parameters on three T. cruzi isolates obtained from wild T. vitticeps specimens. The growth and cell differentiation of the parasite was evaluated through epimastigote and trypomastigote forms obtained in the growth curves for three distinct isolates. The maximum growth showed differences at the 20th day of the curve. Our in vitro results show a heterogeneity, regarding these features for samples cultivated under the same conditions. Morphometric analyzes based on the shape of epimastigotes and trypomastigotes corroborated such differentiation. These results highlight the need of better understanding the meaning of this diversity under an eco-epidemiological perspective792294303COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO CARLOS CHAGAS FILHO DE AMPARO À PESQUISA DO ESTADO DO RIO DE JANEIRO - FAPERJTriatoma vitticeps é um triatomíneo com distribuição geográfica restrita ao território brasileiro, apresentando alta prevalência de infecção natural pelo Trypanosoma cruzi. Esta espécie é relevante sob o ponto de vista epidemiológico por invadir domicílios com frequência nos estados do Rio de Janeiro, Minas Gerais e Espírito Santo. O objetivo deste estudo foi avaliar parâmetros morfológicos e ultraestruturais, em três isolados de T. cruzi obtidos a partir de triatomíneos silvestres. O crescimento e a diferenciação celular do parasita foi avaliado através das formas epimastigotas e tripomastigotas obtidas nas curvas de crescimento para os três isolados. O crescimento máximo mostrou diferenças no 20º dia da curva. Nossos resultados in vitro mostram uma heterogeneidade, em relação a essas características para amostras cultivadas nas mesmas condições. As análises morfométricas baseadas na conformação de epimastigotas e trypomastigotes corroboraram essa diferenciação. Estes resultados ressaltam a necessidade de uma melhor compreensão do significado desta diversidade sob uma perspectiva eco-epidemiológicaThe authors thank “Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Instituto Oswaldo Cruz, Fundação Oswaldo Cruz (IOC-FIOCRUZ), Brasi

Interference With Quorum-Sensing Signal Biosynthesis as a Promising Therapeutic Strategy Against Multidrug-Resistant Pathogens

Faced with the global health threat of increasing resistance to antibiotics, researchers are exploring interventions that target bacterial virulence factors. Quorum sensing is a particularly attractive target because several bacterial virulence factors are controlled by this mechanism. Furthermore, attacking the quorum-sensing signaling network is less likely to select for resistant strains than using conventional antibiotics. Strategies that focus on the inhibition of quorum-sensing signal production are especially attractive because the enzymes involved are expressed in bacterial cells but are not present in their mammalian counterparts. We review here various approaches that are being taken to interfere with quorum-sensing signal production via the inhibition of autoinducer-2 synthesis, PQS synthesis, peptide autoinducer synthesis, and N-acyl-homoserine lactone synthesis. We expect these approaches will lead to the discovery of new quorum-sensing inhibitors that can help to stem the tide of antibiotic resistance

New species of Ehrlichia isolated from Rhipicephalus (Boophilus) microplus shows an ortholog of the E. canis major immunogenic glycoprotein gp36 with a new sequence of tandem repeats

Background: Ehrlichia species are the etiological agents of emerging and life-threatening tick-borne human zoonoses that inflict serious and fatal infections in companion animals and livestock. The aim of this paper was to phylogeneticaly characterise a new species of Ehrlichia isolated from Rhipicephalus (Boophilus) microplus from Minas Gerais, Brazil. Methods: The agent was isolated from the hemolymph of Rhipicephalus (B.) microplus engorged females that had been collected from naturally infested cattle in a farm in the state of Minas Gerais, Brazil. This agent was then established and cultured in IDE8 tick cells. The molecular and phylogenetic analysis was based on 16S rRNA, groEL, dsb, gltA and gp36 genes. We used the maximum likelihood method to construct the phylogenetic trees. Results: The phylogenetic trees based on 16S rRNA, groEL, dsb and gltA showed that the Ehrlichia spp isolated in this study falls in a clade separated from any previously reported Ehrlichia spp. The molecular analysis of the ortholog of gp36, the major immunoreactive glycoproteins in E. canis and ortholog of the E. chaffeensis gp47, showed a unique tandem repeat of 9 amino acids (VPAASGDAQ) when compared with those reported for E. canis, E. chaffeensis and the related mucin-like protein in E. ruminantium. Conclusions: Based on the molecular and phylogenetic analysis of the 16S rRNA, groEL, dsb and gltA genes we concluded that this tick-derived microorganism isolated in Brazil is a new species, named E. mineirensis (UFMG-EV), with predicted novel antigenic properties in the gp36 ortholog glycoprotein. Further studies on this new Ehrlichia spp should address questions about its transmissibility by ticks and its pathogenicity for mammalian hosts

Star Cluster Formation and Disruption Time-Scales -- I. An empirical determination of the disruption time of star clusters in four galaxies

We present a new method to derive the cluster disruption time in selected regions of galaxies from the mass or age distribution of magnitude-limited cluster samples. If the disruption time of clusters in a region of a galaxy depends on their initial mass as t_4 x (M_cluster/10^4 M_sun)^gamma and if the cluster formation rate is constant, then the mass and age distributions of the observed clusters will each show two powerlaw relations. The values of t_4 and gamma can be derived from these relations. We used this method to derive the cluster disruption time in specific regions in four galaxies: the inner region of M51, a region of M33, the SMC and the solar neighbourhood. The values of gamma are the same in the four galaxies within the uncertainty and the mean value is gamma= 0.62 +- 0.06. However the disruption time t_4 of a cluster of 10^4 M_sun is very different in the different galaxies. The clusters in the SMC have the longest disruption time, t_4 = 8 Gyr, and the clusters at 1 to 3 kpc from the nucleus of M51 have the shortest disruption time of t_4 = 0.04 Gyr. The disruption time of clusters 1 to 5 kpc from the nucleus of M33 is t_4 = 0.13 Gyr and for clusters within 1 kpc from the Sun we find t_4 = 1.0 Gyr.Comment: 18 pages, 18 figures. Accepted for publication by Monthly Notice

Tratamiento de la fragilidad y apatía a través de actividades psicomotrices en la enfermedad de Alzheimer

Objective: To implement an Occupational Therapy treatment in elderly people with dementia and frailty Method: A longitudinal study of a sample of 23 users, 14 women and 9 men aged between 71 and 92 is performed, the average age 84.66 years, users of a residence, presenting frailty and dementia. Carried out an initial assessment and final tests with Short Physical Performance Battery (SPPB) and Dementia Apathy Interview and Rating (DAIR), participants attend occupational therapy sessions. Statistical analysis: it was performed nonparametric Wilcoxon, Spearman correlation and Mann Whitney U test. Results: We observed that the levels of fragility do not improve, there is no relationship of age with frailty, the level of apathy does not decrease with the intervention, there is no difference in the score of apathy or fragility by sex. Conclusions: the results indicate a maintenance of vital functions on both scales, which can be considered beneficial when dealing with degenerative processes.Objetivo: Implementar un tratamiento de Terapia Ocupacional en personas mayores con demencia y fragilidad.                                                                                                Método: Se realiza un estudio longitudinal sobre una muestra de 23 usuarios, 14 mujeres y 9 hombres con edades comprendidas entre 71 y 92, siendo la edad media 84.66 años, usuarios de una residencia, que presentan fragilidad y demencia. Se lleva a cabo una evaluación inicial y otra final con las pruebas Batería Corta del Desempeño Físico (SPPB) y Dementia Apathy Interview and Rating (DAIR), los participantes asisten a sesiones de Terapia Ocupacional.        Análisis estadístico: se llevan a cabo pruebas no paramétricas de Wilcoxon, Correlación de Spearman y U de Mann Whitney.                                                                                    Resultados: se observa que los niveles de fragilidad no mejoran, que no hay relación de la edad con la fragilidad, el nivel de apatía no disminuye con la intervención, no existen diferencias en la puntuación de apatía ni fragilidad según el sexo.                                    Conclusiones. los resultados indican un mantenimiento de las funciones vitales en ambas escalas, lo que se puede considerar beneficioso al tratarse de procesos degenerativos

Functional genomics of the horn fly, Haematobia irritans (Linnaeus, 1758)

<p>Abstract</p> <p>Background</p> <p>The horn fly, <it>Haematobia irritans </it>(Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of pastured cattle. Horn flies infestations reduce cattle weight gain and milk production. Additionally, horn flies are mechanical vectors of different pathogens that cause disease in cattle. The aim of this study was to conduct a functional genomics study in female horn flies using Expressed Sequence Tags (EST) analysis and RNA interference (RNAi).</p> <p>Results</p> <p>A cDNA library was made from whole abdominal tissues collected from partially fed adult female horn flies. High quality horn fly ESTs (2,160) were sequenced and assembled into 992 unigenes (178 contigs and 814 singlets) representing molecular functions such as serine proteases, cell metabolism, mitochondrial function, transcription and translation, transport, chromatin structure, vitellogenesis, cytoskeleton, DNA replication, cell response to stress and infection, cell proliferation and cell-cell interactions, intracellular trafficking and secretion, and development. Functional analyses were conducted using RNAi for the first time in horn flies. Gene knockdown by RNAi resulted in higher horn fly mortality (protease inhibitor functional group), reduced oviposition (vitellogenin, ferritin and vATPase groups) or both (immune response and 5'-NUC groups) when compared to controls. Silencing of ubiquitination ESTs did not affect horn fly mortality and ovisposition while gene knockdown in the ferritin and vATPse functional groups reduced mortality when compared to controls.</p> <p>Conclusions</p> <p>These results advanced the molecular characterization of this important ectoparasite and suggested candidate protective antigens for the development of vaccines for the control of horn fly infestations.</p

Geometric deep learning as a potential tool for antimicrobial peptide prediction

Antimicrobial peptides (AMPs) are components of natural immunity against invading pathogens. They are polymers that fold into a variety of three-dimensional structures, enabling their function, with an underlying sequence that is best represented in a non-flat space. The structural data of AMPs exhibits non-Euclidean characteristics, which means that certain properties, e.g., differential manifolds, common system of coordinates, vector space structure, or translation-equivariance, along with basic operations like convolution, in non-Euclidean space are not distinctly established. Geometric deep learning (GDL) refers to a category of machine learning methods that utilize deep neural models to process and analyze data in non-Euclidean settings, such as graphs and manifolds. This emerging field seeks to expand the use of structured models to these domains. This review provides a detailed summary of the latest developments in designing and predicting AMPs utilizing GDL techniques and also discusses both current research gaps and future directions in the field

N-body Models of Extended Clusters

We use direct N-body simulations to investigate the evolution of star clusters with large size-scales with the particular goal of understanding the so-called extended clusters observed in various Local Group galaxies, including M31 and NGC6822. The N-body models incorporate a stellar mass function, stellar evolution and the tidal field of a host galaxy. We find that extended clusters can arise naturally within a weak tidal field provided that the tidal radius is filled at the start of the evolution. Differences in the initial tidal filling-factor can produce marked differences in the subsequent evolution of clusters and the size-scales that would be observed. These differences are more marked than any produced by internal evolution processes linked to the properties of cluster binary stars or the action of an intermediate-mass black hole, based on models performed in this work and previous work to date. Models evolved in a stronger tidal field show that extended clusters cannot form and evolve within the inner regions of a galaxy such as M31. Instead our results support the suggestion many extended clusters found in large galaxies were accreted as members of dwarf galaxies that were subsequently disrupted. Our results also enhance the recent suggestion that star clusters evolve to a common sequence in terms of their size and mass.Comment: 12 pages, 8 figures, accepted by MNRA