RESUMO
BACKGROUND: Drosophila melanogaster activates a variety of immune responses against microbial infections. However, information on the Drosophila immune response to entomopathogenic nematode infections is currently limited. The nematode Heterorhabditis bacteriophora is an insect parasite that forms a mutualistic relationship with the gram-negative bacteria Photorhabdus luminescens. Following infection, the nematodes release the bacteria that quickly multiply within the insect and produce several toxins that eventually kill the host. Although we currently know that the insect immune system interacts with Photorhabdus, information on interaction with the nematode vector is scarce. RESULTS: Here we have used next generation RNA-sequencing to analyze the transcriptional profile of wild-type adult flies infected by axenic Heterorhabditis nematodes (lacking Photorhabdus bacteria), symbiotic Heterorhabditis nematodes (carrying Photorhabdus bacteria), and Photorhabdus bacteria alone. We have obtained approximately 54 million reads from the different infection treatments. Bioinformatic analysis shows that infection with Photorhabdus alters the transcription of a large number of Drosophila genes involved in translational repression as well in response to stress. However, Heterorhabditis infection alters the transcription of several genes that participate in lipidhomeostasis and metabolism, stress responses, DNA/protein synthesis and neuronal functions. We have also identified genes in the fly with potential roles in nematode recognition, anti-nematode activity and nociception. CONCLUSIONS: These findings provide fundamental information on the molecular events that take place in Drosophila upon infection with the two pathogens, either separately or together. Such large-scale transcriptomic analyses set the stage for future functional studies aimed at identifying the exact role of key factors in the Drosophila immune response against nematode-bacteria complexes.
Assuntos
Infecções Bacterianas/genética , Infecções Bacterianas/imunologia , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Infecções por Nematoides/genética , Infecções por Nematoides/imunologia , Photorhabdus/imunologia , Animais , Antibacterianos/imunologia , Biologia Computacional , RNA/genética , Análise de Sequência de RNA/métodos , Transcrição Gênica/genéticaRESUMO
The volatile compounds of five kind of cultivars of jackfruit (Artocarpus heterophyllus Lam.) grown in Nayarit, Mexico, was researched by using extraction and chromatographic methods such as headspace-solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). Eighty-six volatile compounds were identified. The most prominent compounds in the analyzed cultivars were alkyl esters of 3-methylbutanoic acid. Ethyl 3-methylbutanoate was the most abundant ester in FMC, JMC and RMC cultivars (190.7-961.2 µg/kg), whereas butyl 3-methylbutanoate (152.8-205.2 µg/kg) and pentyl 3-methylbutanoate (105.1-210.9 µg/kg) were predominant in DMC and BMC cultivars. By utilizing clustering statistical techniques such as principal component analysis was possible to identify certain esters compounds (number and concentration) to differentiate each cultivar.
Assuntos
Artocarpus , Compostos Orgânicos Voláteis , Cromatografia Gasosa-Espectrometria de Massas , México , Microextração em Fase Sólida , Compostos Orgânicos Voláteis/análiseRESUMO
Background: Alterations in the lipid profile are part of the acute phase response, this corresponds to the so-called lipemia of sepsis. Objective: To determine if the serum level of high density lipoprotein (HDL) is related to severity and mortality. Method: Retrospective, descriptive, cross-sectional study of patients diagnosed with abdominal sepsis. During the period from April 2016 to February 2017. The severity was determined by APACHE II, SOFA, Mannheim, CONUT, the presence of organic faults and mortality. Results: We included 154 cases, 59 female and 95 male; The main organ causing abdominal sepsis was the appendix 41.6%. The overall mortality was 14.3%. The presence of organic faults was 35.1%. The mean HDL level was 37.64 mg/dl (SD ± 16.16). The findings, subjected to statistical verification by Student's t-test, showed significance among the cases with SOFA > 4 (p = 0.01) and Mannheim > 26 (p = 0.001), CONUT > 6 (p = 0.001), presence of organic failures (p = 0.001), and mortality (p = 0.003). Conclusions: HDL levels are related to severity, with the development of organic failures and mortality in sepsis.
Antecedentes: Las alteraciones en el perfil de lípidos son parte de la respuesta de fase aguda, lo que corresponde a la denominada lipemia de la sepsis. Objetivo: Determinar si la concentración sérica de lipoproteínas de alta densidad (HDL) se relaciona con la gravedad y la mortalidad. Método: Estudio retrospectivo, descriptivo, transversal, de pacientes con diagnóstico de sepsis abdominal, durante el periodo de abril de 2016 a febrero de 2017. Se determinó la gravedad mediante APACHE II, SOFA, Mannheim, CONUT, la presencia de fallas orgánicas y la mortalidad. Resultados: Se incluyeron 154 casos, 59 mujeres y 95 hombres. El principal órgano causante de sepsis abdominal fue el apéndice (41.6%). La mortalidad global fue del 14.3%. La presencia de fallas orgánicas fue del 35.1%. El valor medio de HDL se situó en 37.64 mg/dl (desviación estándar: ± 16.16). Los hallazgos, sometidos a verificación estadística mediante la prueba t de Student, mostraron significancia entre los casos con SOFA > 4 (p = 0.01) y Mannheim > 26 (p = 0.001), CONUT > 6 (p = 0.001), presencia de fallas orgánicas (p = 0.001) y mortalidad (p = 0.003). Conclusión: Los valores de HDL se relacionan con la gravedad, con el desarrollo de fallas orgánicas y con la mortalidad en la sepsis.
Assuntos
Gastroenteropatias/sangue , Lipoproteínas HDL/sangue , Sepse/sangue , Índice de Gravidade de Doença , APACHE , Reação de Fase Aguda/sangue , Biomarcadores/sangue , Estudos Transversais , Feminino , Gastroenteropatias/etiologia , Gastroenteropatias/mortalidade , Humanos , Masculino , Pessoa de Meia-Idade , Estado Nutricional , Escores de Disfunção Orgânica , Peritonite/sangue , Peritonite/complicações , Peritonite/mortalidade , Estudos Retrospectivos , Sepse/etiologia , Sepse/mortalidadeRESUMO
BACKGROUND: Procalcitonin is a biomarker of sepsis, whose concentrations increase when some endotoxin enters the bloodstream. It is used, among other things, to discriminate the etiology of infections, increase or decrease the antibiotic spectrum, and predict mortality. OBJECTIVE: To determine the utility of the serum level of procalcitonin as a predictor of severity and mortality. METHOD: Retrospective, descriptive, cross-sectional study of patients diagnosed with abdominal sepsis during the period from April 2016 to February 2017. In all cases the severity was determined by APACHE II, SOFA, Mannheim and CONUT, and mortality. The sample was divided into those with procalcitonin > 10.1 and < 10. RESULTS: We included 99 cases (41 female and 58 male). The main organ causing abdominal sepsis was the appendix 56%. The mean of procalcitonin for the sample was 7.94 (standard deviation: ± 13.76). The findings, subjected to statistical verification by means of the Mann-Whitney U test, showed statistical significance among the cases with procalcitonin 10.1, with the Mannheim scores > 26 points (p = 0.003), CONUT > 6 points (p = 0.027) and presence of organic faults (p = 0.001), but not with APACHE, SOFA and mortality. CONCLUSIONS: Procalcitonin is related to the severity determined by the Mannheim index, CONUT and the development of organic faults.
INTRODUCCIÓN: La procalcitonina es un biomarcador de sepsis, cuyas concentraciones aumentan cuando alguna endotoxina entra en el torrente sanguíneo. Se emplea, entre otras cosas, para discriminar la etiología de las infecciones, escalar/desescalar antibióticos, aumentar o disminuir el espectro antibiotico, y predecir la mortalidad. OBJETIVO: Determinar si los valores de procalcitonina se relacionan con la gravedad y la mortalidad de los pacientes con sepsis abdominal. MÉTODO: Estudio retrospectivo, descriptivo y transversal de pacientes con diagnóstico de sepsis abdominal, de abril de 2016 a febrero de 2017. En todos los casos se determinó la gravedad mediante las escalas APACHE II, SOFA, Mannheim y CONUT, así como la mortalidad. Se dividió la muestra en pacientes con procalcitonina > 10.1 y < 10. RESULTADOS: Se incluyeron 99 casos (41 mujeres y 58 hombres). El principal órgano causante de sepsis abdominal fue el apéndice (56%). La media de procalcitonina para la muestra se situó en 7.94 (desviación estándar: ± 13.76). Los hallazgos, sometidos a verificación estadística mediante la prueba U de Mann-Whitney, mostraron una relación estadísticamente significativa en los casos con procalcitonina 10.1, puntajes de Mannheim > 26 puntos (p = 0.003), CONUT > 6 puntos (p = 0.027) y presencia de fallas orgánicas (p = 0.001); no hubo relación significativa entre los valores de procalcitonina y los puntajes de las escalas APACHE II y SOFA, ni con la mortalidad. CONCLUSIÓN: La gravedad medida por la procalcitonina se relaciona con la gravedad determinada mediante los índices de Mannheim y CONUT, y con el desarrollo de fallas orgánicas.
Assuntos
Pró-Calcitonina/sangue , Sepse/sangue , Sepse/mortalidade , Abdome , Adulto , Biomarcadores/sangue , Estudos Transversais , Feminino , Humanos , Masculino , Peritonite/sangue , Peritonite/complicações , Estudos Retrospectivos , Sepse/etiologia , Índice de Gravidade de DoençaRESUMO
Advanced age is associated with immune system deficits that result in an increased susceptibility to infectious diseases; however, specific mediators of age-dependent immune dysfunction have not been fully elucidated. Here we demonstrated that aged mice exhibit poor effector CD8+ T cell polyfunctionality, primarily due to CD8+ T cell-extrinsic deficits, and that reduced CD8+ T cell polyfunctionality correlates with increased susceptibility to pathogenic diseases. In aged animals challenged with the parasite Encephalitozoon cuniculi, effector CD8+ T cell survival and polyfunctionality were suppressed by highly elevated TGF-ß1. Furthermore, TGF-ß depletion reduced effector CD8+ T cell apoptosis in both young and aged mice and enhanced effector CD8+ T cell polyfunctionality in aged mice. Surprisingly, intrinsic blockade of TGF-ß signaling in CD8+ T cells was sufficient to rescue polyfunctionality in aged animals. Together, these data demonstrate that low levels of TGF-ß1 promote apoptosis of CD8+ effector T cells and high TGF-ß1 levels associated with age result in both CD8+ T cell apoptosis and an altered transcriptional profile, which correlates with loss of polyfunctionality. Furthermore, elevated TGF-ß levels are observed in the elderly human population and in aged Drosophila, suggesting that TGF-ß represents an evolutionarily conserved negative regulator of the immune response in aging organisms.
Assuntos
Envelhecimento/imunologia , Linfócitos T CD8-Positivos/imunologia , Fator de Crescimento Transformador beta1/imunologia , Idoso , Envelhecimento/patologia , Animais , Apoptose/imunologia , Antígenos CD8/genética , Antígenos CD8/imunologia , Linfócitos T CD8-Positivos/classificação , Linfócitos T CD8-Positivos/patologia , Diferenciação Celular/imunologia , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Encephalitozoon cuniculi/imunologia , Encephalitozoon cuniculi/patogenicidade , Feminino , Humanos , Lectinas Tipo C , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Knockout , Modelos Imunológicos , Receptores Imunológicos/metabolismo , Transdução de Sinais/imunologiaRESUMO
Insects have evolved obligate, mutualistic interactions with bacteria without further transmission to other eukaryotic organisms. Such long-term obligate partnerships between insects and bacteria have a profound effect on various physiological functions of the host. Here we provide an overview of the effects of endosymbiotic bacteria on the insect immune system as well as on the immune response of insects to pathogenic infections. Potential mechanisms through which endosymbionts can affect the ability of their host to resist an infection are discussed in the light of recent findings. We finally point out unresolved questions for future research and speculate how the current knowledge can be employed to design and implement measures for the effective control of agricultural insect pests and vectors of diseases.
RESUMO
The cellular arm of the insect immune response is mediated by the activity of hemocytes. While hemocytes have been well-characterized morphologically and functionally in model insects, few studies have characterized the hemocytes of non-model insects. Further, the role of ontogeny in mediating immune response is not well understood in non-model invertebrate systems. The goals of the current study were to (1) determine the effects of caterpillar size (and age) on hemocyte density in naïve caterpillars and caterpillars challenged with non-pathogenic bacteria, and (2) characterize the hemocyte activity and diversity of cell types present in two forest caterpillars: Euclea delphinii and Lithacodes fasciola (Limacodidae). We found that although early and late instar (small and large size, respectively) naïve caterpillars had similar constitutive hemocyte densities in both species, late instar Lithacodes caterpillars injected with non-pathogenic E. coli produced more than a twofold greater density of hemocytes than those in early instars. We also found that both caterpillar species contained plasmatocytes, granulocytes and oenocytoids, all of which are found in other lepidopteran species, but lacked spherulocytes. Granulocytes and plasmatocytes were found to be strongly phagocytic in both species, but granulocytes exhibited a higher phagocytic activity than plasmatocytes. Our results strongly suggest that for at least one measure of immunological response, the production of hemocytes in response to infection, response magnitudes can increase over ontogeny. While the underlying raison d' être for this improvement remains unclear, these findings may be useful in explaining natural patterns of stage-dependent parasitism and pathogen infection.
Assuntos
Borboletas/crescimento & desenvolvimento , Hemócitos/citologia , Larva/crescimento & desenvolvimento , Estágios do Ciclo de Vida , Animais , Contagem de Células Sanguíneas , Tamanho Corporal , Borboletas/citologia , Borboletas/imunologia , Contagem de Células , Tamanho Celular , Larva/citologia , Larva/imunologia , Estágios do Ciclo de Vida/imunologia , Fagocitose/fisiologia , Árvores/parasitologiaRESUMO
Insect hemocytes (equivalent to mammalian white blood cells) play an important role in several physiological processes throughout an insect's life cycle. In larval stages of insects belonging to the orders of Lepidoptera (moths and butterflies) and Diptera (true flies), hemocytes are formed from the lymph gland (a specialized hematopoietic organ) or embryonic cells and can be carried through to the adult stage. Embryonic hemocytes are involved in cell migration during development and chemotaxis regulation during inflammation. They also take part in cell apoptosis and are essential for embryogenesis. Hemocytes mediate the cellular arm of the insect innate immune response that includes several functions, such as cell spreading, cell aggregation, formation of nodules, phagocytosis and encapsulation of foreign invaders. They are also responsible for orchestrating specific insect humoral defenses during infection, such as the production of antimicrobial peptides and other effector molecules. Hemocyte morphology and function have mainly been studied in genetic or physiological insect models, including the fruit fly, Drosophila melanogaster, the mosquitoes Aedes aegypti and Anopheles gambiae and the tobacco hornworm, Manduca sexta. However, little information currently exists about the diversity, classification, morphology and function of hemocytes in non-model insect species, especially those collected from the wild. Here we describe a simple and efficient protocol for extracting hemocytes from wild caterpillars. We use penultimate instar Lithacodes fasciola (yellow-shouldered slug moth) (Figure 1) and Euclea delphinii (spiny oak slug) caterpillars (Lepidoptera: Limacodidae) and show that sufficient volumes of hemolymph (insect blood) can be isolated and hemocyte numbers counted from individual larvae. This method can be used to efficiently study hemocyte types in these species as well as in other related lepidopteran caterpillars harvested from the field, or it can be readily combined with immunological assays designed to investigate hemocyte function following infection with microbial or parasitic organisms.
Assuntos
Hemócitos/citologia , Lepidópteros/química , Animais , Hemócitos/química , Lepidópteros/metabolismo , Microscopia/métodosRESUMO
The aim of this study was to evaluate whether digitized images obtained from occlusal radiographs taken with low or over dose of radiation could be improved with the aid of computer software for digital treatment. Thirteen occlusal radiographs of a dry skull were taken employing 13 different exposure times. The radiographs were digitized and then manipulated with the program for image editing. 143 evaluations were performed by specialists in dental radiology who classified radiographs as appropriate or not appropriate for interpretation. Test Z was used for statistical analysis of the data and the results showed that it is possible to manipulate digitized radiographic images taken with 75% of the ideal exposure time and to make them suitable for interpretation and diagnosis. Conversely, it was concluded that the over exposed images, 57.50% above the standard exposure time, were inadequate.
O objetivo do presente estudo foi verificar a possibilidade de se manipular imagens radiograficas digitalizadas sub e superexpostas e fazer com que as mesmas apresentem boa qualidade para interpretacao e elaboracao do diagnostico radiografico. Foram feitas 13 radiografias intrabucais oclusais totais da maxila em um cranio seco submetido a 13 diferentes tempos de exposicao. As radiografias foram digitalizadas e posteriormente manipuladas em programa de edicao de imagens. Foram realizadas 143 avaliacoes por especialistas em radiologia odontologica que classificaram as imagens em adequadas ou nao para interpretacao. Os resultados mostraram que as imagens sub-expostas quando manipuladas foram consideradas adequadas para interpretacao e elaboracao de diagnostico, o que nao ocorreu com as imagens super-expostas. Sendo assim, concluiu-se que e possivel manipular imagens radiograficas digitalizadas com 75% de reducao do tempo de exposicao ideal e fazer com que as mesmas apresentem qualidade adequada para interpretacao. No entanto, concluiu-se que imagens superexpostas a partir de 57,50% acima do tempo de exposicao padrao ficaram inadequadas para interpretacao e elaboracao do diagnostico.