RESUMO
Entamoeba invadens is the protozoan which causes multiple damages in reptiles and is considered a prototype for the study of the Entamoeba encystment/excystment in vitro. Here we report that EinCerS2 knockdown promoted decrease in sphingomyelin (SM) subspecies with long-chain fatty acids (24:0) down to 50% but increase sphingolipids with short-chain fatty acids (16:0) up to three times in both trophozoites and cysts of E. invadens. EinCerS2 silencing also resulted in decreased trophozoites' movement, proliferation, cysts formation, and trophozoites hatched after excystment. By immunofluorescence assays, a polyclonal antibody against EinCerS2 detected the enzyme in the cytoplasm of E. invadens trophozoites, colocalizing with Endoplasmic Reticulum-resident cognate EiSERCA. Interestingly, EinCerS2 was redistributed close to the plasma membrane during encystation, suggesting that the generation of diacylglycerol (DAG) via synthesis of sphingolipids and the activation protein kinase C might participate in the encystment process of E. invadens.
Assuntos
Movimento Celular , Entamoeba/citologia , Entamoeba/enzimologia , Técnicas de Silenciamento de Genes , Oxirredutases/metabolismo , Trofozoítos/enzimologia , Trofozoítos/crescimento & desenvolvimento , Proliferação de Células , Sobrevivência Celular , Regulação para Baixo/genética , Entamoeba/genética , Amplificação de Genes , Estágios do Ciclo de Vida , Oxirredutases/genética , Transporte Proteico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Esfingomielinas/metabolismoRESUMO
Sphingolipids (SLs) synthesis involves a complex metabolic pathway occurring between the endoplasmic reticulum (ER) and Golgi apparatus, generating ceramide synthesis and complex lipids, respectively. Here we show that E. histolytica, apparently lacking cellular organelles (ER and Golgi apparatus), synthesizes a wide variety of sphingolipid subspecies, being particularly abundant those of long-chain fatty acids. In silico analysis showed five putative genes coding for ceramide synthases (CerS), all of them coding for proteins containing the TLC domain, a region conserved in CerS of multiple organisms. These genes are abundantly expressed in different growth phases. Silencing and overexpression of CerS C4M4U4 (the closest homolog of human CerS 2 and 3) demonstrated its involvement in the synthesis of ceramide. Additionally, we identify C4M4U4, SMS2 and PKC (α, ßII) proteins and their subcellular localization of E. histolytica, suggesting that these subcellular compartments might be involved in the biosynthesis and signaling pathway of sphingolipids, and evidencing different sphingolipid synthesis pathways in Entamoeba.
Assuntos
Entamoeba histolytica/metabolismo , Esfingolipídeos/metabolismo , Vias Biossintéticas , Entamoeba histolytica/genética , Entamebíase/parasitologia , Humanos , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Transdução de Sinais , Esfingolipídeos/genética , Esfingosina N-Aciltransferase/genética , Esfingosina N-Aciltransferase/metabolismoRESUMO
Entamoeba invadens is a protozoan, which causes multiple damages in reptiles and is considered a prototype for the study of the Entamoeba encystment in vitro. Here we report for the first time the role of the de novo synthesis pathway of sphingolipids during the encystment of E. invadens. In silico analysis showed that this parasite has six putative genes coding for ceramide synthases (CerS), all of them coding for proteins containing the Lag1p motif, a region conserved in the ceramide synthases of multiple organisms, suggesting that they might be bona fide CerS. The six genes of E. invadens are differentially expressed at different time intervals in both stages trophozoite and cyst, based on the results obtained through qRT-PCR assays, the genes involved in the synthesis of sphingolipids with long-chain fatty acids CerS 2,3,4 (EIN_046610, EIN_097030, EIN_130350) have maximum points of relative expression in both stages of the E. invadens life cycle, which strongly suggest that the signaling exerted from the synthesis pathway of sphingolipids is essential for the encystment of E. invadens, since the generation of the more abundant sphingomyelin (SM) subspecies with long-chain fatty acids are fundamental for the parasite to reach its conversion from trophozoite to cyst. When myriocin was used as an inhibitor of serine palmitoyl CoA transferase (SPT), first enzyme in the de novo biosynthesis of sphingolipids, the trophozoites of E. invadens were unable to reach the encystment. Since the effect of myriocin was reversed with exogenous d-erythrosphingosine (DHS), it was demonstrated that the inhibition was specific and it was confirmed that the synthesis of sphingolipids play an essential role during the encystment process of E. invadens.
Assuntos
Entamoeba/metabolismo , Encistamento de Parasitas , Esfingolipídeos/metabolismo , Entamoeba/efeitos dos fármacos , Entamoeba/enzimologia , Entamoeba/genética , Ácidos Graxos Monoinsaturados/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Cinética , Estágios do Ciclo de Vida/efeitos dos fármacos , Oxirredutases/genética , Oxirredutases/metabolismo , Encistamento de Parasitas/efeitos dos fármacos , Filogenia , Esfingolipídeos/biossíntese , Esfingomielinas/metabolismo , Esfingosina/análogos & derivados , Esfingosina/farmacologia , Trofozoítos/efeitos dos fármacos , Trofozoítos/genéticaRESUMO
Objetivo: Determinar la mortalidad de dientes permanentes y sus principales causas en usuarios atendidos en las Unidades Comunitarias de Salud Familiar en El Salvador durante el año 2016. Metodología: Esta investigación corresponde a un estudio descriptivo observacional y transversal; el cual describe la mortalidad de dientes permanentes y las principales causas de pérdida, se realizó una cédula de entrevista y una guía de observación a una muestra de 50 personas por cada grupo etario presente, en total 350 por cada investigador, las cuales se realizaron en los municipios de San Fernando, Chalatenango; Nejapa, San Salvador y Oratorio de Concepción, Cuscatlán. Resultados: Se encontró que para los tres municipios en estudio la mayor incidencia de pérdida fue por caries dental, la cual afecta a la población con bajos niveles de escolaridad, seguida por, enfermedad periodontal en adultos, siendo con mayor incidencia de pérdida los dientes anteriores, en tercer lugar se ubica la ausencia por otras causas y trauma dentoalveolar. Conclusiones: El mayor índice de pérdida para la mortalidad indicada y referida es por la enfermedad caries dental, afectando mayormente a personas con escasos recursos económicos y con bajos niveles de escolaridad. La mayoría de sujetos en estudio presentaban al menos la pérdida de una pieza dental permanente a partir del estrato de 16 a 24 años.
Objective: to determine tooth mortality of permanent teeth and its main causes in users attended at Family Health Community Units in El Salvador (UCSF Unidades Comunitarias de Salud Familiar for its acronym in Spanish) during the year of 2016. Methodology: This research corresponds to a descriptive, observational and transversal study; which describes permanent tooth mortality and its main causes of loss; an interview card and an observation guide was made to a sample of 50 people for each age group present, in total 350 for each researcher which were carried on in the municipalities of San Fernando, Chalatenango; Nejapa, San Salvador and Oratorio de Concepcion, Cuscatlán. Results: It was found that for the three municipalities under study, the highest incidence of loss was the absence due to tooth decay that affects the popularity with low schooling levels; followed by periodontal disease in adults, being anterior teeth with more incidence of loss, in third place is the absence due to other causes and finally the absence due to dental trauma. Conclusion: the major indexes of loss, both for reported and indicated mortality due tooth decay, mostly affecting people economically disadvantaged and with low schooling levels. The majority of subjects under study presented loss of a permanent dental piece into the stratum of 16 to 24 years of age.
Assuntos
Dentição Permanente , Dente , Cárie Dentária , El SalvadorAssuntos
Masculino , Feminino , Humanos , Planejamento Familiar , Serviços de Saúde , Condições SociaisRESUMO
A total of 119 unrelated individuals from two of the major ethnic groups in Ecuador were typed for 49 of the autosomal single nucleotide polymorphisms (SNPs) in the SNPforID 52plex using the SNapShot(®) assay. Of the above, 42 samples originated from Mestizos (an admixed population) and the remaining 77 were from Native Amerindian Kichwas. We obtained full SNP profiles in all individuals and concordance of duplicated analyses. No deviation from Hardy-Weinberg equilibrium (HWE) was observed for any SNP in the Mestizo and Kichwa populations and only one and four pairs of loci, respectively showed significant linkage disequilibrium. A relatively low genetic diversity and global positive F(IS) value was observed in Kichwas. A statistically significant global F(ST) value was obtained when the two Ecuadorian populations were compared with populations in Spain, Portugal, Argentina, Denmark, Greenland, China, Somalia and Mozambique. All pairwise F(ST) values were statistically significant. A multi-dimensional scaling based on pairwise F(ST) values showed that the Kichwa population differed from all other populations investigated and that the Mestizos had an intermediate position between Kichwas and Europeans. An admixture analysis indicated that the greater contributor to the Mestizo population was the Kichwas (71.2%) compared to the European contribution. The combined mean match probability and mean paternity exclusion probability were 3.3 × 10(-17) and 0.998, respectively, for the Mestizo population and 3.3 × 10(-14) and 0.993, respectively, for the Kichwa population.
Assuntos
Etnicidade/genética , Genética Populacional , Polimorfismo de Nucleotídeo Único , Impressões Digitais de DNA , Equador , Frequência do Gene , Humanos , Desequilíbrio de Ligação , Reação em Cadeia da PolimeraseRESUMO
Studying the Y chromosomes of indigenous tribes of Ecuador revealed a lack of strategic SNP assays to examine the substructure of South American native populations. In most studies dealing with South American samples so far only the most common Y-SNP M3 of haplogroup Q was analyzed, because this is known to define a founder group in South America. Studies of SNPs ancestral to Q-M3 (Q1a3a) to confirm the results or the typing of Q subclades have often been neglected. For this reason we developed a SNaPshot assay, which allows first for a hierarchical testing of all main haplogroups occurring in South American populations and second for a detailed analysis of haplogroups Q and C thought having ancient Asian descent. We selected 16 SNPs from the YCC haplogroup tree and established two multiplexes. The first multiplex ("SA Major") includes 12 Y-SNPs defining the most frequent haplogroups occurring in South America (M42, M207, M242, M168, M3, M145, M174, M213, RPS4Y711, M45, P170, and M9). The second multiplex ("SA SpecQ") contains Y-SNPs of haplogroup Q, especially of the subclade Q-M3 (M19, M194, P292, M3, and M199). Within our Ecuadorian sample, haplogroup Q-M3 (xM19, M194, P292, and M199) was predominant, but we also found haplogroup E and R, which can be attributed to recent admixture. Moreover, we found four out of 65 samples, which were tested to be haplogroup C3* (C-M217) the modal haplogroup in Mongolians and widespread in indigenous populations of the Russian Far East as well as in Eastern Asia. This haplogroup is not known to be the result of recent admixture and has been found only one time before in South America. Since haplogroup C occurs in Asia and in North America (C3b or C-P39), we assume that these C-lineages are ancient as well. Therefore, we established a third multiplex ("SA SpecC"), which allows the further subtyping of haplogroup C, mainly of subclade C3 defined by the Y-SNP M217 (M407, M48, P53.1, M217, P62, RPS4Y711, M93, M86, and P39). Altogether, these three multiplexes cover the most frequent haplogroups in South America and allow for a maximal resolution of the Y-chromosomal SNP diversity in Amerindian population samples.
Assuntos
Cromossomos Humanos Y , Etnicidade/genética , Genética Populacional , Polimorfismo de Nucleotídeo Único , Impressões Digitais de DNA , Equador , Eletroforese Capilar , Haplótipos , Humanos , Masculino , Filogenia , Filogeografia , Reação em Cadeia da Polimerase , Sequências de Repetição em TandemRESUMO
Adipogenesis is regulated by a complex cascade of transcriptional factors, but little is known about the early events that regulate the adipogenic program. Here, we report the role of the srebf1a gene in the differentiation of fibroblastic 3T3-F442A cells. We found that expression of srebf1a depended on GSK3ß activity and that GSK3ß activity was necessary for C/EBPß phosphorylation at Thr188. Knockdown of srebf1a inhibited the adipogenic program because it blocked the expression of genes encoding PPARγ2, C/EBPα, SREBP1c and even FABP4, demonstrating that SREBP1a activation is upstream of these three essential adipogenic transcription factors. Kinetic analysis during differentiation illustrated that the order of expression of adipogenic genes was the following: cebpb, srebf1a, pparg2, cebpa, srebp1c and fabp4. Our data suggest that srebf1a acts as an essential link between the GSK3ß-C/EBPß signaling axis and the beginning of the adipogenic transcriptional cascade.
Assuntos
Adipogenia , Regulação da Expressão Gênica , Quinase 3 da Glicogênio Sintase/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/fisiologia , Células 3T3 , Adipócitos/citologia , Tecido Adiposo/citologia , Animais , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Diferenciação Celular , Glicogênio Sintase Quinase 3 beta , Camundongos , PPAR gama/metabolismo , Fenótipo , Fosforilação , Isoformas de Proteínas/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Transcrição GênicaRESUMO
Three main ethnic groups live in the South American country of Ecuador: Mestizos, Amerindian natives, and African-derived populations, or Afro-Ecuadorans. Mestizos and Afro-Ecuadorans can be considered trihybrid populations containing genes originating in the Americas, Europe, and Africa, as is the case with equivalent populations in other Latin American countries. The proportion and the dynamics of the admixture process remain unknown. However, a certain sex asymmetry of the admixture process can be expected for historical reasons. We typed 11 Y-chromosome short tandem repeats (STRs) in these three ethnic groups to provide adequate allele and haplotype frequencies for forensic genetic purposes and to quantify admixture proportions in male lineages. In addition, a data set of 15 autosomal STRs in the same samples were reanalyzed for the same purpose. Contributions to Mestizo Y chromosomes were estimated to be 70% European, 28% Amerindian, and 2% African, whereas in autosomes the contributions were 19%, 73%, and 8%, respectively, which underlines the sexual asymmetry in mating, with Europeans contributing mostly males. European Y-chromosome haplotypes in Mestizos were similar to those in Spain. Moreover, about 10% of European Y chromosomes were found in the Amerindian Kichwa. As for Afro-Ecuadorans, their contributions to the male line are 44% African, 31% European, and 15% Native American; the last value is the highest percentage reported so far for an African-derived American group. Autosomal admixture was estimated as 56% African, 16% European, and 28% Amerindian.
Assuntos
Cromossomos Humanos Y/genética , Etnicidade/genética , Genética Populacional/métodos , Haplótipos/genética , Repetições de Microssatélites/genética , Equador , Feminino , Humanos , MasculinoAssuntos
Competência Clínica , Pessoal de Saúde/educação , Promoção da Saúde/métodos , Acessibilidade aos Serviços de Saúde , Vasectomia , Adulto , Guatemala , Conhecimentos, Atitudes e Prática em Saúde , Pessoal de Saúde/economia , Hospitais , Humanos , Masculino , Avaliação de Processos em Cuidados de Saúde , Desenvolvimento de Programas , Administração em Saúde Pública/economia , Administração em Saúde Pública/métodos , Vasectomia/métodos , Vasectomia/normas , Vasectomia/estatística & dados numéricosAssuntos
Humanos , Masculino , Feminino , Adulto , Angioplastia Coronária com Balão/métodos , Angioplastia Coronária com Balão , Ecocardiografia Transesofagiana/métodos , Ecocardiografia Transesofagiana , Ecocardiografia/métodos , Ecocardiografia , Anastomose de Artéria Torácica Interna-Coronária/métodos , Microcirculação/anormalidades , Revascularização Miocárdica/métodos , StentsRESUMO
La litotripsia con ondas de choque extracorpóreas, es un recurso poco disponible que ha demostrado ser un elemento útil en el manejo de la colédocolitiasis difícil, cuando se combina con métodos endoscópicos, sus resultados son satisfactorios, con porcentajes de éxito global del 70 al 80 por ciento y con una baja morbilidad y mortalidad.
Assuntos
Humanos , Masculino , Feminino , Idoso , Colangiografia/estatística & dados numéricos , Endoscopia , Doenças Biliares/terapia , Cálculos Biliares/terapia , Litotripsia/estatística & dados numéricosRESUMO
Expone que el Lupus Eritematoso fue descrito en 1851 por Sir William Osler, como una enfermedad autoinmune. Se la ha llamado "la enfermedad de las mil caras" debido a que con frecuencia su diagnóstico se vuelve difícil en etapas iniciales. El LES comienza frecuentemente en los años fértiles de la mujer, sin embargo, se ha descrito en individuos de ambos sexos de cualquier raza o nacionalidad, desde los 2 hasta los 97 años de edad. Se realizó un estudio retrospectivo de 53 pacientes con LES, en base a un análisis documental de los datos exsitentes en las historias clínicas, en un período de 3 años. La muestra fue tomada de 3 hospitales de Quito, se consideraron solo aquellas que tenían nota de ingreso, los análisis de laboratorio de rutina completo y la epicrisis. Se seleccionaron 31 variables, entre signos y síntomas y test laboratoriales para su estudio. Los resultados más importantes fueron: se encontró un 33.3 por ciento de los pacientes entre los 31 y 40 años; el 92 por ciento de los estudiados fueron mujeres, de las cuáles un 55 por ciento ha estado embarazada y ha tenido un parto cefalovaginal en un 78 por ciento de los casos...