Triacylglycerol biosynthesis occurs via the glycerol-3-phosphate pathway in the insect Rhodnius prolixus

Abstract Although triacylglycerol (TAG) stores play a critical role in organisms, mechanisms underlying TAG synthesis are poorly understood in invertebrates. In mammals, the synthesis of glycerolipids, including TAG, diacylglycerol (DAG) and phospholipids (PL), occurs predominantly by the glycerol-3-phosphate (G3P) pathway in most cell types, except for in enterocytes. In these cells, the monoacylglycerol (MAG) pathway accounts for the majority of glycerolipid production. The insect Rhodnius prolixus, a vector of Chagas' disease, exhibits a high capacity to produce glycerolipids in the midgut after a blood meal, providing substrates that are transferred to other organs, such as the fat body, which is specialized in TAG production and storage. In this report, the genes required for TAG synthesis were identified in the R. prolixus genome. The genomic data indicated that TAG is synthesized by the G3P pathway, which is the sole pathway for TAG synthesis in this organism. Furthermore, transcription of both the RpGpat and RpDgat genes were upregulated in a diverse number of organs at moments of highest lipid production. In the midgut and fat body, in vitro synthesis of glycerolipids required G3P, but not MAG, as the initial substrate. These results indicate that the G3P pathway is the only route for TAG synthesis in R. prolixus, and its regulation at the transcriptional level can be a determinant of glycerolipid synthesis and TAG formation in insect organs

Identification of uncoupling protein 4 from the blood-sucking insect Rhodnius prolixus and its possible role on protection against oxidative stress

Abstract Uncoupling proteins (UCPs) play a critical role in the control of the mitochondrial membrane potential (ΔΨm) due to their ability to dissipate the proton gradient, which results in the uncoupling of mitochondrial respiration from ATP production. Most reactive oxygen species generation in mitochondria occurs in complex III, due to an increase of semiquinone (Qradical dot−) half-life. When active, UCPs can account as a potential antioxidant system by decreasing ΔΨm and increasing mitochondrial respiration, thus reducing Qradical dot− life time. The hematophagous insect Rhodnius prolixus, a vector of Chagas disease, is exposed to a huge increase in oxidative stress after a blood meal because of the hydrolysis of hemoglobin and the release of the cytotoxic heme molecule. Although some protective mechanisms were already described for this insect and other hematophagous arthropods, the putative role of UCP proteins as antioxidants in this context has not been explored. In this report, two genes encoding UCP proteins (RpUcp4 and RpUcp5) were identified in the R. prolixus genome. RpUcp4 is the predominant transcript in most analyzed organs, and both mRNA and protein expression are upregulated (13- and 3-fold increase, respectively) in enterocytes the first day after the blood feeding. The increase in UCP4 expression is coincident with the decrease in hydrogen peroxide (H2O2) generation by midgut cells. Furthermore, in mitochondria isolated from enterocytes, the modulation of UCP activity by palmitic acid and GDP resulted in altered ΔΨm, as well as modulation of H2O2 generation rates. These results indicate that R. prolixus UCP4 may function in an antioxidation mechanism to protect the midgut cells against oxidative damage caused by blood digestion

Looking for reference genes for real-time quantitative PCR experiments in Rhodnius prolixus (Hemiptera: Reduviidae)

Abstract Quantitative real-time PCR (qPCR) has become one of the most used techniques to measure gene expression. However, normalization of gene expression data against reference genes is essential, although these are usually used without any kind of validation. The expression of seven genes was compared in organs of Rhodnius prolixus under diverse conditions, using published software to test gene expression stability. Rp18S and elongation factor 1 (RpEF -1) were the most reliable genes for normalization in qPCR when gene expression in different organs was compared. Moreover, both genes were found to be the best references when transcript levels were compared in the posterior midgut of insects infected with Trypanosoma cruzi. Rp18S was also the best reference gene in the fat bodies of unfed and fed insects. By contrast, RpEF-1 was found to be the best reference gene for comparison between posterior midguts, and RpMIP or RpActin should be used to compare gene expression in the ovaries. Although Rp18S is indicated here as the best reference in most cases, reports from the literature show that it is difficult to find an optimum reference gene. Nevertheless, validation of candidate genes to be taken as references is important when new experimental conditions are tested to avoid incorrect data interpretation

Long-chain acyl-CoA synthetase 2 knockdown leads to decreased fatty acid oxidation in fat body and reduced reproductive capacity in the insect Rhodnius prolixus

Long-chain acyl-CoA esters are important intermediates in lipid metabolism and are synthesized from fatty acids by long-chain acyl-CoA synthetases (ACSL). The hematophagous insect Rhodnius prolixus, a vector of Chagas' disease, produces glycerolipids in the midgut after a blood meal, which are stored as triacylglycerol in the fat body and eggs. We identified twenty acyl-CoA synthetase genes in R. prolixus, two encoding ACSL isoforms (RhoprAcsl1 and RhoprAcsl2). RhoprAcsl1 transcripts increased in posterior midgut on the second day after feeding, and RhoprAcsl2 was highly transcribed on the tenth day. Both enzymes were expressed in Escherichia coli. Recombinant RhoprACSL1 and RhoprACSL2 had broad pH optima (7.5-9.5 and 6.5-9.5, respectively), were inhibited by triacsin C, and were rosiglitazone-insensitive. Both showed similar apparent Km for palmitic and oleic acid (2-6 μM), but different Km for arachidonic acid (0.5 and 6 μM for RhoprACSL1-Flag and RhoprACSL2-Flag, respectively). The knockdown of RhoprAcsl1 did not result in noticeable phenotypes. However, RhoprACSL2 deficient insects exhibited a 2.5-fold increase in triacylglycerol content in the fat body, and 90% decrease in fatty acid β-oxidation. RhoprAcsl2 knockdown also resulted in 20% increase in lifespan, delayed digestion, 30% reduced oviposition, and 50% reduction in egg hatching. Laid eggs and hatched nymphs showed remarkable alterations in morphology. In summary, R. prolixus ACSL isoforms have distinct roles on lipid metabolism. Although RhoprACSL1 functions remain unclear, we propose that RhoprACSL2 is the main contributor for the formation of the intracellular acyl-CoA pool channeled for β-oxidation in the fat body, and is also required for normal reproduction

Serotonin regulates an acyl-CoA-binding protein (ACBP) gene expression in the midgut of Rhodnius prolixus

Abstract Acyl-CoA esters have many intracellular functions, acting as energy source, substrate for metabolic processes and taking part in cell signaling. The acyl-CoA-binding protein (ACBP), a highly conserved 10 kDa intracellular protein, binds long- and medium-chain acyl-CoA esters with very high affinity, directing them to specific metabolic routes and protecting them from hydrolysis. An ACBP gene sequence was identified in the genome of Rhodnius prolixus. This ACBP gene (RpACBP-1) was expressed in all analyzed tissues and quantitative PCR showed that expression was highest in posterior midgut. In this tissue, ACBP gene expression increased in the first day after blood meal (∼10-fold) and then decreased to unfed levels in the seventh day after meal. Injection of serotonin (5-hydroxytryptamine; 5-HT), a neuroamine released in the hemolymph after the start of feeding, increased the expression of this gene in the midgut of unfed females, reaching levels similar to those observed in fed insects. This effect of injected 5-HT was inhibited by spiperone, an antagonist of 5-HT mammalian receptors, that was also able to block the physiological increase in RpACBP-1 expression observed after feeding. Injection of cholera toxin or dibutyryl-cAMP also resulted in the stimulation of this gene expression. These data reveal a transcriptional regulatory mechanism in R. prolixus, that is triggered by 5-HT. In this way, a novel role for 5-HT is proposed, as a regulator of ACBP gene expression and, consequently, taking part in the control of lipid metabolism

Metabolic syndrome in elderly from a northeastern brazilian city

Introduction: Population aging is a global reality. In Brazil, it is so expressive and in 2050 is estimated that the number of men and women over 80 years old can overcome the 20-24 years old population and also children under 14 years old. Metabolic syndrome is defined as a set of cardiovascular risk factors, detection and intervention in this age group may decrease cardiovascular mortality. We aimed to estimate the prevalence of metabolic syndrome (MS) in elderly patients in a northeastern Brazilian city. Method: This is a quantitative, observational, cross-sectional and population-based study. The population consisted of non-institutionalized individuals aged> 60 years old, attended in the Family Health Strategy (FHS) and residents in Cajazeiras, PB, Brazil. For the diagnosis of metabolic syndrome we considered the criteria recommended by the International Federation of Diabetes. Results: We studied 351 elderly patients with 72.4+8.7 years old and 66.1% were female. The prevalence of MS was 69.8% and the most frequent criterion was increased waist circumference (82.6%), followed by high triglycerides (81.5%) and low HDL-cholesterol (80.6%). We observed a statistically significant positive association between MS and women [OR: 1.38 (95% CI: 1.18 to 1.63)] and housing in urban areas [OR: 1.23 (1.02 to 1.48)]. Conclusion: There is a high prevalence of MS among the elderly mainly in women and those living in urban areas

INTERVENÇÕES EFICAZES NA REDUÇÃO DA MORTALIDADE MATERNO-INFANTIL: UMA ANÁLISE ABRANGENTE

ABSTRACT Introduction: Maternal and child mortality remains a significant public health challenge around the world, especially in developing countries. The main causes include complications during pregnancy, childbirth and postpartum, such as bleeding, infections and hypertension. Objectives: This study will review the interventions used to reduce maternal and child mortality and describe their effectiveness and importance. Methodology: Research carried out in March 2024, based on an integrative review of scientific literature in the SciELo database. Using the descriptors: "Maternal and child mortality" and "Reduction in mortality". Results and discussion: Interventions used to reduce maternal and child mortality include adequate prenatal care, birth assistance by qualified health professionals, access to effective contraceptive methods, childhood immunization, promotion of breastfeeding and improved access to health services. health. Conclusion: The reviewed interventions offer effective and fundamental strategies for reducing maternal and child mortality. Implementation of these measures, combined with ongoing efforts to promote equitable access to health services, is essential. With targeted investments, we can advance the protection of the health of women and children around the world, ensuring that every pregnancy and birth is safe and healthy.  Introdução: A mortalidade materno-infantil continua sendo um desafio significativo de saúde pública em todo o mundo, especialmente em países em desenvolvimento. As principais causas incluem complicações durante a gravidez, parto e pós-parto, como hemorragias, infecções e hipertensão. Objetivos: Esse estudo vida revisar as intervenções utilizadas na redução da mortalidade materno-infantil e descrever a eficácia e importância das mesmas. Metodologia: Pesquisa realizada em março de 2024, com base em uma revisão integrativa da literatura científica na base de dados SciELo. Utilizando os descritores: "Mortalidade materno-infantil" e "Redução da mortalidade". Resultados e discussão: As intervenções utilizadas para reduzir a mortalidade materno-infantil incluem cuidados pré-natais adequados, assistência ao parto por profissionais de saúde qualificados, acesso a métodos contraceptivos eficazes, imunização infantil, promoção do aleitamento materno e melhoria do acesso a serviços de saúde. Conclusão: As intervenções revisadas oferecem estratégias eficazes e fundamentais na redução da mortalidade materno-infantil. A implementação dessas medidas, combinada com esforços contínuos para promover acesso equitativo aos serviços de saúde, é essencial. Com investimentos direcionados, podemos avançar na proteção da saúde de mulheres e crianças em todo o mundo, garantindo que cada gravidez e nascimento seja seguro e saudável

Pais/responsáveis como coadjuvantes na saúde bucal de seus filhos

To verify the level of knowledge of parents and guardians about dentistry in early childhood, and to analyze sociodemographic variables. This was a field research, observational and cross-sectional with quantitative approach and descriptive and analytical analysis of the data, through questionnaires, which were applied to parents / guardians of children from 0 to 3 years old from municipal daycare centers a city of the Paraíba hinterland. 171 parents or guardians of children enrolled in municipal public day care centers were interviewed. Most of them were predominant females (91.2%), aged 15 to 30 years (63.2%), with monthly family income of up to R $780.00 (67.3$ 780,00 (67,3%), y tenía un nivel de educación predominante en la escuela primaria I (33.9%). La mayoría de los padres / tutores (55.6%) consideraron que la salud oral significaba higiene y eran conscientes de la existencia de odontología para bebés (64.9%). Para el 52.6% de los encuestados, el momento ideal para comenzar la higiene de la cavidad bucal del bebé es desde el nacimiento del primer diente y en el método utilizado para realizar esta higiene, el 53.8% indicó gasa o pañal empapado en agua filtrada. Se observó que el 53.8% informó que la importancia de la higiene oral está relacionada con la salud general del bebé. Los padres y / o tutores de los niños de guarderías públicas demostraron un conocimiento razonable sobre la salud bucal de los niños.Verificar o nível de conhecimento dos pais e responsáveis sobre a odontologia na primeira infância, além de analisar variáveis sociodemográficas. Tratou-se de uma pesquisa de campo, observacional e transversal com abordagem quantitativa e análise descritiva e analítica dos dados, por meio de questionários, que foram aplicados a pais/responsáveis por crianças de 0 a 3 anos de idades de creches municipais de uma cidade do sertão paraíbano. Foram entrevistados 171 pais ou responsáveis por crianças matriculadas em creches públicas de um município do sertão paraibano. Observou-se que a maioria era predominante do gênero feminino (91,2%), com 15 à 30 anos de idade (63,2%), possuíam renda mensal familiar de até R$ 780,00 (67,3%), e tinham nível de escolaridade predominante o fundamental I (33,9%). A maioria dos pais/ responsáveis (55,6%), consideraram que saúde bucal significava higiene e tinham conhecimento sobre a existência da Odontologia para bebês (64,9%). Para 52,6% dos entrevistados, o momento ideal iniciar a higiene da cavidade bucal do bebê é a partir do nascimento do primeiro dente e sobre o método empregado para realizar essa higienização, 53,8% indicaram gaze ou fralda embebida em água filtrada. Foi observado que 53,8% relataram que a importância da higiene bucal está relacionada saúde geral do bebê. Pais e/ou responsáveis por bebês das creches públicas demonstraram um conhecimento razoável, em relação à saúde bucal infantil

Syzygium jambolanum treatment improves survival in lethal sepsis induced in mice

<p>Abstract</p> <p>Background</p> <p>The leaves and the fruits from <it>Syzygium jambolanum </it>DC.(Myrtaceae), a plant known in Brazil as sweet olive or 'jambolão', have been used by native people to treat infectious diseases, diabetes, and stomachache. Since the bactericidal activity of <it>S. jambolanum </it>has been confirmed <it>in vitro</it>, the aim of this work was to evaluate the effect of the prophylactic treatment with <it>S. jambolanum </it>on the <it>in vivo </it>polymicrobial infection induced by cecal ligation and puncture (CLP) in mice.</p> <p>Methods</p> <p>C57Bl/6 mice were treated by the subcutaneous route with a hydroalcoholic extract from fresh leaves of <it>S. jambolanum </it>(HCE). After 6 h, a bacterial infection was induced in the peritoneum using the lethal CLP model. The mice were killed 12 h after the CLP induction to evaluate the cellular influx and local and systemic inflammatory mediators' production. Some animals were maintained alive to evaluate the survival rate.</p> <p>Results</p> <p>The prophylactic HCE treatment increased the mice survival, the neutrophil migration to infectious site, the spreading ability and the hydrogen peroxide release, but decreased the serum TNF and nitrite. Despite the increased migration and activation of peritoneal cells the HCE treatment did not decrease the number of CFU. The HCE treatment induced a significant decrease on the bone marrow cells number but did not alter the cell number of the spleen and lymph node.</p> <p>Conclusion</p> <p>We conclude that the treatment with <it>S. jambolanum </it>has a potent prophylactic anti-septic effect that is not associated to a direct microbicidal effect but it is associated to a recruitment of activated neutrophils to the infectious site and to a diminished systemic inflammatory response.</p

Relato de experiência sobre consulta de enfermagem realizada por acadêmicos como estratégia de promoção de saúde / Experience report on the nursing consultation performed by students as a health promotion strategy

Objetivo: Apresentar as experiências e as atividades vividas por acadêmicos de enfermagem sob supervisão de um professor, compartilhando aspectos teóricos e práticos do momento vivido pelos autores neste singular ambiente de assistência de enfermagem como forma de promoção em saúde para prevenção de diabetes mellitus e hipertensão arterial sistêmica. Métodos: Trata-se de um relato de experiência, com característica qualitativa a partir de métodos descritivos e observacionais. Foi realizada uma ação educativa pelos acadêmicos de enfermagem juntamente de um professor, com 93 participantes, em um Centro Universitário localizado na cidade de Niterói/RJ, em maio de 2022. Criou-se um instrumento para aplicação na consulta de enfermagem. Resultados e discussão: A realização das consultas de enfermagem oportunizou o amadurecimento profissional dos graduandos, visto que estes exerceram o raciocínio clínico ao pôr em prática seus conhecimentos teóricos e assistenciais previamente adquiridos no curso. Além disso, o público assistido ressaltou a importância do acolhimento recebido nas consultas, sendo um diferencial na prática do autocuidado. Conclusão: Os acadêmicos identificaram a potencialidade do enfermeiro diante do seu papel como educador em saúde. Diante disso, sugere-se construir consultórios de enfermagem nas universidades ao entender a sua importância na formação do profissional enfermeiro, oportunizando vivências práticas ricas em aprendizado ao reconhecer a realidade do aumento das doenças crônicas não transmissíveis no país