Validation of reference genes for expression analysis in the salivary gland and the intestine of Rhodnius prolixus (Hemiptera, Reduviidae) under different experimental conditions by quantitative real-time PCR

<p>Abstract</p> <p>Background</p> <p><it>Rhodnius prolixus </it>is a blood-feeding insect that can transmit <it>Trypanosoma cruzi </it>and <it>Trypanosoma rangeli </it>to vertebrate hosts. Recently, genomic resources for invertebrate vectors of human pathogens have increased significantly, and <it>R. prolixus </it>has been one of the main species studied among the triatomines. However, the paucity of information on many of the fundamental molecular aspects of this species limits the use of the available genomic information. The present study aimed to facilitate gene expression studies by identifying the most suitable reference genes for the normalization of mRNA expression data from qPCR.</p> <p>Results</p> <p>The expression stability of five candidate reference genes (<it>18S </it>rRNA, <it>GAPDH</it>, β-actin, α-tubulin and ribosomal protein <it>L26</it>) was evaluated by qPCR in two tissues (salivary gland and intestine) and under different physiological conditions: before and after blood feeding and after infection with <it>T. cruzi </it>or <it>T. rangeli</it>. The results were analyzed with three software programs: geNorm, NormFinder and BestKeeper. All of the evaluated candidate genes proved to be acceptable as reference genes, but some were found to be more appropriate depending on the experimental conditions. <it>18S</it>, <it>GAPDH </it>and α-tubulin showed acceptable stability for studies in all of the tissues and experimental conditions evaluated. β-actin, one of the most widely used reference genes, was confirmed to be one of the most suitable reference genes in studies with salivary glands, but it had the lowest expression stability in the intestine after insect blood feeding. <it>L26 </it>was identified as the poorest reference gene in the studies performed.</p> <p>Conclusions</p> <p>The expression stability of the genes varies in different tissue samples and under different experimental conditions. The results provided by three statistical packages emphasize the suitability of all five of the tested reference genes in both the crop and the salivary glands with a few exceptions. The results emphasise the importance of validating reference genes for qRT-PCR analysis in <it>R. prolixus </it>studies.</p

Application of RNA interference in triatomine (Hemiptera: Reduviidae) studies.

Triatomines (Hemiptera: Reduviidae) are obligate hematophagous insects. They are of medical importance because they are vectors of Trypanosoma cruzi, the causative agent of Chagas disease in the Americas. In recent years, the RNA interference (RNAi) technology has emerged as a practical and useful alternative means of studying gene function in insects, including triatomine bugs. RNAi research in triatomines is still in its early stages, several issues still need to be elucidated, including the description of the molecules involved in the RNAi machinery and aspects related to phenotype evaluation and persistence of the knockdown in different tissues and organs. This review considers recent applications of RNAi to triatomine research, describing the major methods that have been applied during the knockdown process such as the double-stranded RNA delivery mechanism (injection, microinjection, or ingestion) and the phenotype characterization (mRNA and target protein levels) in studies conducted with the intent to provide greater insights into the biology of these insects. In addition to the characterization of insect biomolecules, some with biopharmacological potential, RNAi may provide a new view of the interaction between triatomine and trypanosomatids, enabling the development of new measures for vector control and transmission of the parasite

Paradigmas em psicologia: compreensões acerca da saúde e dos estudos epidemiológicos Paradigms in psychology: comprehension about health and epidimiology studies

O presente artigo tem como objetivo investigar de que forma a saúde é entendida pelos diferentes paradigmas em psicologia e como são avaliados os estudos epidemiológicos. Para tanto, entrevistou-se sete pesquisadores e profissionais da área da psicologia do Brasil, Espanha e Argentina, que orientam suas práticas pelos seguintes referenciais teóricos e metodológicos: social-crítico, clínico (enfoque psicodinâmico e cognitivo-comportamental), ecológico-contextual e comunitário. A partir dos dados obtidos na investigação empírica observou-se que há uma diversidade de compreensões do conceito de saúde. O conceito de saúde apresentado pelos diferentes paradigmas está relacionados à elementos como a concepção de homem, mundo, realidade, influência do contexto social e possibilidade de mudança. Em relação aos estudos epidemiológicos nota-se que alguns participantes voltam-se a uma descrição superficial e ampla, vinculada à concepção médica do conceito de epidemiologia; outros ressaltam a relevância de se considerar os contextos em que são realizados estes estudos. Apenas no um participante mencionou o papel da epidemiologia especificamente para psicologia. Concluímos que é de fundamental importância que o profissional da saúde tenha consciência do paradigma que orienta sua prática para poder avaliar as conseqüências de sua atuação como profissional e cidadão.<br>This article aims is investigate how health is understood by differents paradigms in psychology and how they evaluated epidemiology studies. Seven investigators and psychology health professional from Brazil, Spain and Argentina were interviews, this professionals guide their practices by differents conceptual theories: Social Critique, Clinical (Psychodynamic and Cognitive Behavior), Ecological Contextual and Community. After the evaluation of those interviews it was possible identify that exist a diversity of health compression. The health concepts are related with the human, world, reality, social context and possibility of change compression concepts. In relation to the epidemiology studies we realizes that some participations use a superficial and large description, related to the medical conception about the epidemiology; others emphasizes the relevance of to consider the contexts that the studies are achieves. Only one participant mentioned the role of epidemiology specifically for psychology. We conclude that is of fundamental importance that the professional of health has conscience of the paradigm that directs your practice can to evaluate the consequences of your action like professional and citizen

Inflammatory Response and Activation of Coagulation after COVID-19 Infection

SARS-CoV-2 (COVID-19) infection is responsible for causing a disease with a wide spectrum of clinical presentations. Predisposition to thromboembolic disease due to excessive inflammation is also attributed to the disease. The objective of this study was to characterize the clinical and laboratory aspects of hospitalized patients, in addition to studying the pattern of serum cytokines, and associate them with the occurrence of thromboembolic events. Methodology: A retrospective cohort study with 97 COVID-19 patients hospitalized from April to August 2020 in the Triângulo Mineiro macro-region was carried out. A review of medical records was conducted to evaluate the clinical and laboratory aspects and the frequency of thrombosis, as well as the measurement of cytokines, in the groups that presented or did not present a thrombotic event. Results: There were seven confirmed cases of thrombotic occurrence in the cohort. A reduction in the time of prothrombin activity was observed in the group with thrombosis. Further, 27.8% of all patients had thrombocytopenia. In the group that had thrombotic events, the levels of IL1b, IL-10, and IL2 were higher (p < 0.05). Conclusions: In the studied sample, there was an increase in the inflammatory response in patients with thrombotic events, confirmed by the increase in cytokines. Furthermore, in this cohort, a link was observed between the IL-10 percentage and an increased chance of a thrombotic event