187 resultados para Giardia lamblia
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核仁是真核细胞核中最明显的结构,是rRNA转录、加工和核糖体亚基组装的场所。核仁的起源进化无疑是真核细胞起源进化问题的一个极为重要的方面。但迄今为止,这方面的研究很少。本文首先对"不具核仁"的低等单细胞真核生物贾第虫的"核仁"基因及核糖体合成系统的基因群进行了研究,进而对已知的核仁蛋白质组的生物进行了比较基因组研究,从而从基因和基因组的角度对核仁的起源进化问题进行了较为深入的研究。获得了如下结,和结论:1)在"不具核仁"的贾第虫中分离鉴定出了典型真核细胞参与pre-rRNA加工的krr]基因;表明贾第虫中的pre-rRNA加工机制与典型真核细胞的是一致的。在具有核仁结构的典型真核细胞中,krr1基因所编码的KRRIP蛋白是参与Pre一rRNA加工的核仁定位蛋白之一。我们在一度被认为是极为原始的"不具核仁结构"的真核生物-贾弟虫的基因组中分离鉴定出了该基因,并证明该基因在贾第虫中是活跃转录的。同时,还调查发现与KRRIP形成蛋白复合体从而发挥功能的众场,其基因在贾第虫中也是存在的,这提示贾第虫的pre寸RNA加工机制与具有核仁结构的典型真核细胞是一致的。2)对贾第虫核糖体合成系统的基因群/亚基因组进行了全面调查,发现在典型真核生物中所共有的参与核搪体合成的129条保守蛋白基因中有89条在贾第虫基因组具有相应的直系同源基因,包括编码参与rRNA申基化和假尿嗜陡化的蛋白复合体成员以及存在于905、405和605复合体中的蛋白等诸多重要基因。这表明贾第虫的核糖体合成系统与典型的真核生物相似,只是参与的成分相对简单些。以上1)、2)的结果表明贾第虫虽然没有核仁结构,但其核糖体合成机制却与具核仁结构的典型真核细胞一致。这可能意味着真核细胞核仁的核糖体合成功能的起源形成是在核仁结构形成之前,那么核仁结构的形成可能是在此功能之外另有"新意";另外也可能是由于贾第虫因适应寄生生活而导致核仁"退化"之故,虽然这种可能性偏少,但若果真如此也是生物适应性进化中的一个重要现象,值得深入研究。3)对已知核仁蛋白质组的人、酵母和拟南芥进行了比较基因组学研究。通过搜索KOG直系同源蛋白簇数据库,发现约74%的人的核仁蛋白,约75%的酵母的核仁蛋白以及约84%的拟南芥的核仁蛋白在动物、真菌和植物中都保守。这表明这些高等真核生物的核仁蛋白大都在它们分化之前就已起源形成。进一步的调查发现:共同定位于人和酵母核仁中的同源蛋白中,人有154条蛋白对应于酵母的134条蛋白,其中人的核仁蛋白相对于酵母的有10条蛋白发生了基因重复;同样的分析发现,人的核仁蛋自相对于拟南芥的有21条蛋白发生了基因重复。这些事实表明:随着真核生物的进化,至少在高等真核生物中基因重复是核仁成分复杂化和核仁进化的重要途径。4)用人的442条核仁蛋白(它们在植物和真菌中也都存在同源蛋白,但不一定是核仁蛋白)对原核生物基因组进行了搜索。结果表明,在真细菌和原(古)细菌中都能找到好多同源物,但发现下列重要现象:只在原(古)细菌有同源物的核仁蛋白要比只在真细菌中有同源物的核仁蛋白多得多,包括了RNA修饰蛋白、核糖体蛋自以及参与翻译的相关蛋白等;而只在真细菌中有同源物的核仁蛋白只是RNA螺旋酶和WD重复蛋白。因此,我们认为核仁可能是由真细菌和原细菌复合起源的,但其主体起源于原细菌。
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MicroRNAs (miRNAs)是一类长约21-25nt 的非编码小分子RNAs,通过与靶基因的互补结合在转录水平及转录后水平来负调控基因表达。人们已在众多高等多细胞生物中如人、果蝇、线虫、拟南芥等鉴定出众多microRNAs 分子。近来报道单细胞原生生物衣藻中也存在大量microRNAs。然而到目前为止,在被很多证据证实是最原始的单细胞真核生物贾第虫中却仍未有microRNAs 的报道。那么到底贾第虫这种具有特殊进化地位的单细胞原生动物是否存在有microRNAs 呢?如果存在的话,其microRNAs 的特点是什么?与高等多细胞生物及单细胞衣藻的 microRNA 相比又有何异同点呢?贾第虫的microRNAs 是否与其致病性相关呢?已有研究表明,贾第虫基因组中存在与RNAi 相关的Argonaute(AGO)家族蛋白和Dicer 酶。有意思的是,这些与siRNA 引起RNAi 作用关键的蛋白AGO 和Dicer 同样也是miRNA 系统的关键成份,这就提示我们在贾第虫中很有可能也存在有miRNA 并发挥功能。有研究发现在贾第虫基因组中存在大量的非编码转录物,这些大量的非编码转录物中,是否都是后来所认为的为双向启动子转录有用基因时的副产物,还是也存在一些起调控作用的RNA 分子(如miRNAs 等),需要进一步的研究。本文利用生物信息学的手段,依据miRNAs 的生物学特征,结合多种计算机预测的方法,在贾第虫基因组中筛选可能的microRNAs 分子,结果共鉴定出50 个miRNAs 候选分子,这50 个可能的贾第虫miRNAs 不具有保守性,在已知的其他物种的miRNAs 中找不到同源物。用这50 个microRNAs BLASTN 贾第虫的蛋白质编码序列及其相邻5’端和3’端各200bp 的序列,来寻找这些microRNAs 所调控的靶基因。结果表明,寻找到的贾第虫miRNA 的靶基因除很大一部分未知功能的蛋白外,还包括了很多涉及不同功能的蛋白,如VSP 蛋白(various surface proteins)这样一类表面抗原蛋白,提示我们贾第虫miRNA 可能与其致病性相关。接下来我们对其中14 个预测的贾第虫microRNAs 进行了RT-PCR 检测并克隆测序,结果表明gla-mir-6, gla-mir-35 在贾第虫滋养体细胞中稳定表达。我们的研究第一次用生物信息学结合实验的方法在贾第虫寻找到了microRNAs,为下一步深入研究这些microRNAs 在贾第虫中的功能提供了可能。
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典型的真核生物有四种rRNA(18S、5.8S、28S和5SrRNA)。一般18S、5.8S和28S的基因分别由转录间隔区(ITS)隔开而位于同一个转录单位上构成一个rRNA基因拷贝,多个rRNA基因拷贝串联形成rDNA。rDNA聚集在一起构成核仁组织区(NOR),成为核仁发生的位置。5SrRNA基因除在少数真核生物(如:酵母)中是和18S、28S rRNA基因位于同一个转录单位上外,一般是处在核仁以外的区域。贾第虫一度被认为是现存最原始的真核生物。支持这一观点的一个重要证据之一就是它还不具核仁结构。那么它的rDNA与典型真核生物的相比会有怎样的特点呢?本文在基因组的水平上对贾第虫的rDNA进行了全面调查分析,并对5S rRNA及其相关蛋白进行重点研究,得到如下结果和结论: 1)贾第虫的18S rRNA(1448bp)基因和28S rRNA(2300bp)基因比其他一些真核生物的(一般为1800bp和3400bp)要小的多,甚至比一些原核生物的相应的rRNA基因还要小。不仅如此,其5.8S rRNA基因和28SrRNA基因之间的转录间隔区(ITS2)比典型真核生物的对应区域也要短得多(只有54bp),且GC含量较高。结构预测表明该间隔区不能形成在许多真核生物中所能形成的保守的二级结构。更特别的是,贾第虫基因组中的rRNA基因序列大部分都是不完整的,并且不按照18S-5.8S-28S rRNA基因顺序排列,也没有多个完整拷贝顺序排列的区域。这提示贾第虫rRNA基因可能是以一种不同于典型真核生物的方式聚集的。因此本文认为以上这些特点可能与贾第虫不能形成典型核仁结构有关。 2)本文从贾第虫基因组中鉴定出了5S rRNA基因,并实验验证了其表达及其完整基因序列所编码的5S rRNA具有典型真核生物的T型二级结构,且具有绝大多数保守位点。RT-PCR表明该基因具有转录活性。该结果否定了前人的贾第虫没有5S rRNA的实验结果。并表明贾第虫尽管很原始,但其5S rRNA基因仍然是独立存在的和单独转录的。贾第虫基因组中总共有8个5S rRNA基因拷贝(且其中还有一个拷贝具有15个bp的异常插入)这大大低于一般真核生物的拷贝数。这些5S rRNA基因也不形成串联排列的区域。 我们还在贾第虫中鉴定出在真核生物中唯一与5S rRNA接触的核糖体蛋白L5蛋白并验证了其表达,该序列与其他真核生物的L5蛋白相似性很高,这提示贾第虫在5S rRNA基因转录出核后与L5蛋白结合形成5S RNP的过程可能与典型的真核生物是一致的。此外,我们从贾第虫中鉴定不出符合典型真核生物TFIIIA因子特征的蛋白,这提示贾第虫5S rRNA的转录起始以及转录后出核的机制可能与典型真核生物不同。过去对贾第虫的研究表明高等真核生物里RNA聚合酶III所独有的四个亚基在贾第虫中找不到同源物,而这样不完整的RNA聚合酶III已经可以在贾第虫中完成5S rRNA的转录了,这表明RNA聚合酶III所独有的这些亚基可能是为了完成其他功能而进化出来的。
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Previously we suggested that four proteins including aldolase and triose phosphate isomerase (TPI) evolved with approximately constant rates over long periods covering the whole animal phyla. The constant rates of aldolase and TPI evolution were reexamined based on three different models for estimating evolutionary distances, It was shown that the evolutionary rates remain essentially unchanged in comparisons not only between different classes of vertebrates but also between vertebrates and arthropods and even between animals and plants, irrespective of the models used, Thus these enzymes might be useful molecular clocks for inferring divergence times of animal phyla, To know the divergence time of Parazoa and Eumetazoa and that of Cephalochordata and Vertebrata, the aldolase cDNAs from Ephydatia fluviatilis, a freshwater sponge, and the TPI cDNAs from Ephydatia fluviatilis and Branchiostoma belcheri an amphioxus, have been cloned and sequenced, Comparisons of the deduced amino acid sequences of aldolase and TPI from the freshwater sponge with known sequences revealed that the Parazoa-Eumetazoa split occurred about 940 million years ago (Ma) as determined by the average of two proteins and three models, Similarly, the aldolase and TPI clocks suggest that vertebrates and amphioxus last shared a common ancestor around 700 Ma and they possibly diverged shortly after the divergence of deuterostomes and protostomes.
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A homogenous detection of pathogen (Giardia lamblia cysts) based on the catalytic growth of gold nanoparticles (AuNPs) has been studied. In this study, centrifugal filters were employed as tools to concentrate and separate the pathogen cells, and moreover amplify the detection signal. The catalytic growth of gold nanoparticles was verified to be positively related to gold seeds concentration. On this basis, homogenous detection of the pathogenic bacteria in liquid phase was established by means of conjugating antibody to gold seeds. Under the given experimental condition, detection limit of G. lamblia cysts was determined as low as 1.088 × 103 cells ml-1. The additional nonspecific binding tests were also conducted to verify the detection specificity. This sensing platform has been proved to be a sensitive, reliable and simple method for large-scale pathogen detection, and provide valuable insight for the development of gold nanocrystals based colorimetric biosensors.
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Introdução: Os parasitas intestinais são responsáveis por morbilidade em crianças de todo mundo, em especial nos países de baixa renda. Os estudos têm vindo a demonstrar o seu impacto negativo no estado nutricional e o seu contributo na etiologia da anemia. Pretendeu-se determinar a prevalência de parasitas intestinais em crianças dos 5 aos 12 anos de idade, a frequentar a escola primária no Bairro Lucrécia, no Lubango, Angola, e explorar a sua relação com o estado nutricional e anemia. Material e Métodos: Foi efectuado um estudo observacional, transversal e analítico, cuja colheita de dados se realizou entre Setembro e Outubro de 2010. A amostra foi constituída por 328 crianças. Realizou-se a detecção microscópica de parasitas intestinais e identificação molecular dos parasitas Entamoeba histolytica e Entamoeba dispar. O estado nutricional foi avaliado através dos z-scores do peso para a idade, da estatura para a idade e do IMC para a idade. A concentração de hemoglobina foi determinada através de um hemoglobinómetro portátil. Resultados: A prevalência de parasitas intestinais patogénicos foi de 44,2%, destacando-se Ascaris lumbricoides com 22,0%, Giardia lamblia com 20,1% e Hymenolepis nana com 8,8%. Na microscopia foi encontrada uma prevalência de Entamoeba histolytica/dispar de 13,7%, tendo sido posteriormente identificada, por diagnóstico molecular, uma prevalência de 13,1% para E. dispar e 0,3% para E. histolytica. A prevalência de baixo peso, subnutrição crónica e subnutrição aguda foi de, respectivamente, 36,1%, 41,5% e 30,2%. A probabilidade das crianças terem subnutrição crónica ou subnutrição aguda aumentou com o facto de terem 10 anos ou mais. As crianças co-infectadas por protozoários e helmintas apresentaram uma maior probabilidade de terem subnutrição crónica. A prevalência de anemia foi de 21,6%, encontrando-se a mesma significativamente associada à infecção por H. nana. A probabilidade das crianças estarem anémicas aumentou com o facto de terem menos de 10 anos. Adicionalmente nas crianças desparasitadas com albendazol ou mebendazol há 2 meses e meio ou menos verificou-se uma maior prevalência de infecção por G. lamblia (28,6%) em comparação com as desparasitadas há mais de 2 meses e meio (13,7%), tendo sido essa diferença estatisticamente significativa. Discussão e Conclusões: Emergiu deste estudo a importância da co-infecção com helmintas e protozoários no aumento da probabilidade das crianças terem subnutrição crónica e foi encontrada uma associação estatisticamente significativa entre a infecção por H. nana e a anemia. Será importante desenhar futuros estudos que investiguem o poder patogénico do H. nana e o modo como é efectuada a desparasitação com albendazol ou mebendazol, pois ao ser eficaz contra a infecção por A. lumbricoides, poderá aumentar a susceptibilidade à infecção por G. lamblia.
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Affiliation: Pascal Michel : Département de pathologie et microbiologie, Faculté de médecine vétérinaire
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Infections for intestinal parasites are one of the main morbidade causes in humans and, its relationships with socioeconomic levels and hygiene conditions in countries in development are already very established. Many works, even so, they are being accomplished to elucidate the complex interactions among nutrition, these infections and answer imunológica, because it is seen that malnutrition commits the immunity increasing the susceptibilidade for infectious diseases and these for its time can harm the state human nutricional. It is known that sponge helmínticos they stimulate synthesis of IgE so much policlonal as specific for the same ones and that IgA secretora, main imunoglobulina of defense of the mucous ones, can act against protozoa as the Giardia lamblia and against helmintos as Trichuris tichiura and Strongyloides stercorales. Some studies show that the malnutrition energy protéica influences in the production of these answers, but some authors show results divergentes. In this work it was evaluated the levels of total IgE, IgA sérica and secretora, contagem of sanguine eosinófilos, levels of proteins séricas and state nutricional, in 103 children of low socioeconomic level, to discover a correlation between those and infection for enteroparasitas. They participated in the study children of both sexes, with age of 3 to 6 years, visitors of the same creche and residents in a neighborhood with precarious hygiene conditions and basic saneamento, in the city of Christmas. The obtained results showed that the faulty environmental and socioeconomic conditions favored to a high infection frequency for enteroparasitas, mainly Trichuris trichiura and Ascaris lumbricoides between the helmintos and Endolimax sleep and Giárdia lamblia among the protozoa. Light malnutrition without deficit protéico was observed in 30% of the children, which didn't also present significant deficiencies of IgA sérica and secretora. The sponged children
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The exclusive breastfeeding in the first six months of life practice is essential for the health and development of children. Studies show there is in human milk protective antibodies against intestinal parasites and a relationship between the absence of breastfeeding or weaning and parasitic infections. This work was a prospective cohort study involving 34 pregnant adolescents attended school in Maternity Januario Cicco / RN and their children, to assess the influence of breastfeeding on intestinal parasites in them. Thus, the parasitic infection was investigated by examination of feces parasitological and environmental factors by use of questionnaires. The average age of the mothers studied was 16.2 years. Of these 76.4% were infected and the most prevalent species of parasites were Entamoeba histolytica/dispar (76.9%), Giardia lamblia (19.2%) and Ascaris lumbricoides (11.5%). The infected children had an average age of 5.1 months and the frequency of parasites was 61.7%. The infection was earlier detecctada with 45 days of life. The most common parasites were Giardia lamblia (40%), Entamoeba histolytica/dispar (35%), Ascaris lumbricoides (5%) and Ancylostoma (5%). The average length of exclusive breastfeeding was 2.2 months. There was an association between increased duration of exclusive breastfeeding and increased time to detection of parasites in the feces of children. There was no statistically significant correlation between the socio-economic conditions and cultural and breastfeeding. The data reinforce the importance of breastfeeding to protect the children in its first year of life
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Em um mundo de grande pobreza, as enteroparasitoses podem exercer um papel importante no que se refere às desigualdades sociais, pois estas mazelas se apresentam, em sua grande maioria, na população mais carente. Estas mesmas doenças causam inúmeros casos de faltas ao trabalho e a escola, gerando preocupação dos profissionais de saúde, já que estes parasitas podem agravar ou desencadear quadros de anemia e desnutrição. O trabalho procurou pesquisar a prevalência de enteroparasitas na Mesorregião do Oeste Potiguar, em uma população que utiliza o sistema público de saúde, buscando associar as variáveis, como: sexo, faixa etária, período sazonal, renda familiar e número de parasitas. Foi utilizada a técnica de Hofmann (sedimentação espontânea) e o material fecal foi analisado pelos funcionários do Laboratório Central de Mossoró e Pau dos Ferros. Foi realizado no período de 02 de março de 2010 a 28 de março de 2011 o exame coprológico em 16.246 pessoas no laboratório Central de Mossoró (Lacen) e Pau dos Ferros, RN. A análise dos exames mostrou que 69,23% dos infectados eram mulheres e 33,07% eram homens. A maior prevalência ocorreu com Endolimax nana (41,61%), seguido de Entamoeba coli (18,46%) e Giardia lamblia (18,59%) em 3.407 parasitas encontrados. Dentre os exames realizados, o Endolimax nana foi o mais prevalente entre os organismos encontrados. A diferença nos resultados entre mulheres e homens pode determinar um foco domiciliar, pelo descaso do poder público com as políticas de saneamento básico e falta de higiene da população, e pela atividade das mesmas com trabalhos domésticos
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This study applied a socioeconomic questionnaire designed to evaluate the frequency of intestinal parasites and characterize epidemiological, nutritional, and immunological variables in 105 HIV/AIDS patients - with and without parasitic infections, attending the Day Hospital in Botucatu, UNESP, from 2007 to 2008. Body mass index was calculated and the following tests performed: parasitological stool examinations; eosinophil, IgE, CD4(+) T and CD8(+) T lymphocyte cell counts; albumin test; viral load measure; and TNF-alpha, IFN-gamma, IL-2, IL-5 and IL-10 cytokine levels. Results were positive for parasitic intestinal infections in 12.4% of individuals. Most patients had good socioeconomic conditions with basic sanitation, urban dwellings, treated water supply and sewage, good nutritional and immunological status and were undergoing HAART. Parasites were found at the following frequencies: Entamoeba - five patients (38.5%), Giardia lamblia-four (30.7%), Blastocystis hominis-three (23.0%), Endolimax nana-two (15.4%), and Ascaris lumbricoides - one (7.7%). There were no significant differences between the two groups for eosinophils, albumin, IgE, CD4(+) T and CD8(+) T lymphocytes, INF-gamma, IL-2, or IL-10. Most patients also showed undetectable viral load levels. Significant differences were found for TNF-alpha and IL-5. These results show the importance of new studies on immunodeficient individuals to increase understanding of such variables.
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A sample of 471 pre-school children who frequented schools and creches in a poor district of Manaus (Amazonas), Brazil, were randomly submitted to faecal parasitological tests. Two-hundred-and-forty children from both sexes between the ages of 3 and 7 years with Ascaris lumbricoides and/or Giardia lamblia were selected. The objective of the study was to determine the possible influence of these two intestinal parasites and vitamin A and/or zinc supplementation on the serum retinol levels of primary school children. The children were submitted to clinical and anthropometric examinations, dietary interviews and biochemical examinations of retinol and carotene in the serum and of zinc in the hair. The parasitic incidence was 85.0% and about 54% of the children were polyparasitic. During the pretreatment phase, the retinol and carotene serum levels were 36% and 57%, respectively, below the normal levels. Using the Waterlow classification, the anthropometric analyses revealed that 88% of the children showed normal growth. A significant effect was observed of the anti-parasitic medicine on the serum retinol levels.
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The objective of this study was to verify the prevalence of intestinal parasites and/or commensals in the neoplastic patients undergoing chemotherapy. Stool samples were analyzed by the method of Lutz (1919) and Rugai (1954), in triplicate. This work was composed of three groups, the first one (GI) formed by neoplastic patients that are not undergoing chemotherapy, the second (GII) comprised patients who were undergoing chemotherapy, and the third group (GIII) consisting of patients who completed chemotherapy. A total of 30 patients (GI-5, GII-18 and GIII-7) were screened at the Assis Regional Hospital of the Unified Health System of Assis, São Paulo. Additional information on antiparasitic treatment and tumor type were obtained by questionnaire. The positivity was 66.7% (20 cases) for intestinal parasites and/or commensals. The helminths were Ascaris lumbricoides (36.7%), Hookworms (20%) and Hymenolepis diminuta (3.3%). Among the highlights are protozoan Giardia lamblia (46.7%), Entamoeba coli (6.7%), E. histolytica/dispar (3.3%), Endolimax nana (3.3%) and Iodameba butschlii (3.3%). The high frequency of intestinal parasites and/or commensals in the neoplastic patients can be attributed to poor personal hygiene and lack of immunity to reinfection and poor knowledge of the prophylaxis of infection by protozoa and helminths. The results indicate the necessity of adopting a new criterion for neoplastic patients undergoing chemotherapy, primarily performing parasitological diagnosis, treatment and monitoring of cure of intestinal parasitic infections in this risk group.
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Considering the high prevalence of the parasitic diseases in Brazil and its close relationship with the socio-economic and cultural levels of the population, aimed to establish the occurrence of enteroparasites in children of six Centers of Education Child (CEIs) of the municipality of Patos of Minas (MG). Were collected three samples of faeces of 161 children from zero to six years of age. Were used the Lutz and Baermann-Moraes' methods, to the detection of cysts of protozoa, eggs and larvae of helminths. Coproparasitological analyses showed positivity of 73%. The most frequent helminths were: Ascaris lumbricoides (50%), hookworm (22%) and Enterobius vermicularis (0.6%). Among the protozoa highlighted were: Entamoeba histolytica/E. dispar (22%) and Giardia lamblia (32%). The high occurrence of enteroparasites evidenced the need for an effective health policy in the CEIs of the municipality of Patos of Minas to the fight against intestinal parasites.
Resumo:
Infestation by protozoa and/or helminths is considered to be extremely rare in infants. We therefore reviewed the records of all infants up to 12 mth of age attending the Paediatric Department from January 1973 to June 1977, in order to assess the frequency of stool examinations for parasites within this age group, the percentage of positive results in these examinations, and some personal characteristics which propitiated parasitic infestation. Some characteristics of the investigated infants were compared to those of a representative sample of the total number of infants attending during the same period. Stool examinations were realized in 11.8% of the 1,162 attending patients and of those, 15.3% were positive. The proportion of positive cases became 24.0% when the infants in whom there was a history of parasites being passed were included. Ascaris lumbricoides and Giardia lamblia were the parasites most frequently found, respectively in 42.8% and 37.1% of the positive cases. The only detected difference between the control group and the group of infants who had parasitologic examinations was the presence of diarrhea, significantly more frequent in the investigated group. The proportion of positive cases was significantly greater in girls, in infants older than 6 mth and when diarrhea was present for up to 15 days, than in boys, in infants up to 6 mth of age and in chronic diarrhea, respectively. Our results show that parasites were investigated in only a small percentage of infants under 1 yr of age, but that in this age group already, parasitic infestation does occur with a certain frequency. As investigated cases were selected, mainly due to the presence of diarrhea, the real prevalence of parasitism in infants under 1 yr of age was impossible to evaluate.