Asexual stage

  • 文章类型: Journal Article
    背景:恶性疟原虫疟疾是一种主要在热带国家出现的公共卫生问题。直到现在,目前还没有针对恶性疟原虫感染血液阶段特异性抗原的有效疟疾疫苗。因为疟疾的发病机制是血液阶段感染的结果,在自然暴露于疟疾感染的情况下,确定最有希望的血液阶段疫苗候选抗原至关重要。
    方法:在南贝宁实施了一个由400名孕妇及其婴儿组成的队列。在怀孕和婴儿期建立了主动和被动的疟疾监测方案,以准确确定随访期间的疟疾感染情况。在妊娠(3个时间点)和婴儿期(6个时间点)重复定量对7种疟疾疫苗候选抗原具有特异性的28种抗体(Ab)反应,以研究Ab动力学及其保护作用。Abs通过ELISA和逻辑定量,线性和cox比例风险模型分析了Ab反应与母亲和婴儿对疟疾的保护之间的关联,考虑到社会经济因素和婴儿的环境暴露风险。
    结果:从生命的第18个月开始,针对MSP1、MSP2和MSP3的IgM水平显示出针对有症状的疟疾感染发作的早期保护性反应,而婴儿期未发现IgG应答相关。在女性中,一些IgG反应往往与在怀孕期间和分娩时预防疟疾风险有关,其中IgG3抗GLURP-R0和IgG2抗MSP1。
    结论:主要发现表明,在婴儿时期的疫苗设计中应考虑IgM。研究IgM在疟疾保护中的功能作用需要进一步关注。
    BACKGROUND: Plasmodium falciparum malaria is a public health issue mostly seen in tropical countries. Until now, there is no effective malaria vaccine against antigens specific to the blood-stage of P. falciparum infection. Because the pathogenesis of malarial disease results from blood-stage infection, it is essential to identify the most promising blood-stage vaccine candidate antigens under natural exposure to malaria infection.
    METHODS: A cohort of 400 pregnant women and their infants was implemented in South Benin. An active and passive protocol of malaria surveillance was established during pregnancy and infancy to precisely ascertain malaria infections during the follow-up. Twenty-eight antibody (Ab) responses specific to seven malaria candidate vaccine antigens were repeatedly quantified during pregnancy (3 time points) and infancy (6 time points) in order to study the Ab kinetics and their protective role. Abs were quantified by ELISA and logistic, linear and cox-proportional hazard model were performed to analyse the associations between Ab responses and protection against malaria in mothers and infants, taking into account socio-economic factors and for infants an environmental risk of exposure.
    RESULTS: The levels of IgM against MSP1, MSP2 and MSP3 showed an early protective response against the onset of symptomatic malaria infections starting from the 18th month of life, whereas no association was found for IgG responses during infancy. In women, some IgG responses tend to be associated with a protection against malaria risk along pregnancy and at delivery, among them IgG3 against GLURP-R0 and IgG2 against MSP1.
    CONCLUSIONS: The main finding suggests that IgM should be considered in vaccine designs during infanthood. Investigation of the functional role played by IgM in malaria protection needs further attention.
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  • 文章类型: Journal Article
    类霉菌,由Mollisia(Fr。)P.岩溶。,其特点是柔软,无柄的带球形的,深色细胞切除,透明子囊孢子,和全球分布在温带地区。由于缺乏独特的特征,它们的通用和物种划界很困难,迫切需要基于DNA序列的研究。两个属的类真菌,BelonopsisandTrichobelonium,包含相对较少的物种,并被(0-)1-3-间隔子囊孢子识别,髓外由松散交织的菌丝组成,和草酸钙晶体。从日本的各个地点收集了形态上可分配给Belonopsis或Trichobelonum的未描述物种的标本,并尝试将其分配给适当的属。根据基于ITS串联序列的莫利西尔科成员的分子系统发育分析,LSU,和RPB1,八个分类实体被放置在具有Mollisiadiesbachiana的强烈支持的单枝中,从Belonopsis的类型物种中分离出来,B.excelsior.因此,提出了一个新的新属以适应未描述的物种。在这项研究中,描述了8个新物种的新叶白藻和2个新物种的毛叶白藻。还为M.diesbachiana提出了一种新的组合。分子分析支持了Neobelonopsis和Trichobelonium的一般区别。确定了一些其他特征来界定毛比隆,例如在大脑皮层和下腺之间存在锚定菌丝,以及在菌落上产生丰富的晶体和可溶性色素。发现新球藻的衍生物种在子囊孢子中具有多间隔。
    Mollisioid fungi, represented by Mollisia (Fr.) P. Karst., are characterized by soft, sessile apothecia with globose, dark-celled excipula, hyaline ascospores, and worldwide distribution in temperate regions. Their generic and species delimitation is difficult due to the lack of distinct features, and studies based on DNA sequences are urgently required. Two genera of mollisioid fungi, Belonopsis and Trichobelonium, comprise relatively few species and are recognized by (0-)1-3-septate ascospores, medullary excipulum composed of loosely interwoven hyphae, and calcium oxalate crystals in the excipulum. Specimens of undescribed species that are morphologically assignable to Belonopsis or Trichobelonium were collected from various sites in Japan and their assignment to the proper genera was attempted. According to a molecular phylogenetic analysis involving members of Mollisiaceae based on concatenated sequences of ITS, LSU, and RPB1, eight taxonomic entities were placed in a strongly supported single clade with Mollisiadiesbachiana, separated from the type species of Belonopsis, B.excelsior. A new genus Neobelonopsis was thus proposed to accommodate the undescribed species. In this study, eight new species of Neobelonopsis and two new species of Trichobelonium were described. A new combination was also proposed for M.diesbachiana. The generic distinction of Neobelonopsis and Trichobelonium was supported by molecular analysis. Some additional characteristics to delimit Trichobelonium were identified, such as the presence of anchoring hyphae between the base of the apothecium and subiculum, and the production of abundant crystals and soluble pigments on the colonies. Derivative species of Neobelonopsis were found to have multi-septa in ascospores.
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  • 文章类型: Journal Article
    背景:吉卜索尼巴贝斯,犬巴贝斯虫病的病原体,属于顶孔门。体外培养技术的发展推动了各种组学研究的进展,包括疟原虫的转录组学分析。在体外和体内环境之间,这促进了诊断抗原的观察和疫苗的开发。然而,没有关于巴贝西亚的信息。可以在这方面获得,这极大地阻碍了对寄生虫在血液阶段的生长发育的进一步了解。
    方法:在本研究中,与体内寄生虫相比,观察到连续体外培养的B.gibsoni(武汉分离株)的形态和感染性发生了很大变化。基于这些变化,从体内和体外培养物中收集B.gibsoni(武汉分离株),然后进行总RNA提取和Illumina转录组测序。获得的差异表达基因(DEGs)使用qRT-PCR进行验证,然后通过几个数据库进行功能注释。从B.gibsoni(武汉分离株)的基因组中克隆了体外培养后表达最高的基因,并通过Western印迹和间接免疫荧光测定法进行了表征,以检测天然形式和细胞定位。
    结果:通过实验室培养,观察到多种形式的寄生虫,发现体外培养的寄生虫在狗中的感染性较低。基于这些变化,进行了Illumina转录组测序,显示377个单基因上调,334个单基因下调。值得注意的是,AP2转录因子家族,对寄生虫的所有发育阶段都至关重要,被筛选,并测试了这些家族成员的转录变化。因此,选择体外适应后上调表达最高的新型AP2转录因子基因(BgAP2-M)。该基因包含1989个碱基对的开放阅读框(ORF),其编码662个氨基酸的全长蛋白质。BgAP2-M包含一个AP2域和一个ACDC保守域,这可能与寄生虫的核生物学有关。制备的针对BgAP2-M肽的多克隆抗体进一步检测到〜73kDa的天然大小,并定位到吉布氏芽孢杆菌的细胞核。
    结论:本研究首次在体内和体外对双歧杆菌进行了全面的转录组分析,有助于详细了解环境变化对血液阶段寄生虫生长和发育的影响。此外,它还为ApiAP2转录因子家族的不同成员作为Babesiaspp的各种生命阶段调节因子提供了更深入的研究。
    BACKGROUND: Babesia gibsoni, the causative agent of canine babesiosis, belongs to the phylum Apicomplexa. The development of in vitro culture technology has driven research progress in various kinds of omics studies, including transcriptomic analysis of Plasmodium spp. between in vitro and in vivo environments, which has prompted the observation of diagnostic antigens and vaccine development. Nevertheless, no information on Babesia spp. could be obtained in this respect, which greatly hinders the further understanding of parasite growth and development in the blood stage.
    METHODS: In this study, considerable changes in the morphology and infectivity of continuous in vitro cultured B. gibsoni (Wuhan isolate) were observed compared to in vivo parasites. Based on these changes, B. gibsoni (Wuhan isolate) was collected from both in vivo and in vitro cultures, followed by total RNA extraction and Illumina transcriptome sequencing. The acquired differentially expressed genes (DEGs) were validated using qRT-PCR, and then functionally annotated through several databases. The gene with the greatest upregulation after in vitro culture was cloned from the genome of B. gibsoni (Wuhan isolate) and characterized by western blotting and indirect immunofluorescence assay for detecting the native form and cellular localization.
    RESULTS: Through laboratory cultivation, multiple forms of parasites were observed, and the infectivity of in vitro cultured parasites in dogs was found to be lower. Based on these changes, Illumina transcriptome sequencing was conducted, showing that 377 unigenes were upregulated and 334 unigenes were downregulated. Notably, an AP2 transcription factor family, essential for all developmental stages of parasites, was screened, and the transcriptional changes in these family members were tested. Thus, the novel AP2 transcription factor gene (BgAP2-M) with the highest upregulated expression after in vitro adaptation was selected. This gene comprises an open reading frame (ORF) of 1989 base pairs encoding a full-length protein of 662 amino acids. BgAP2-M contains one AP2 domain and one ACDC conserved domain, which may be involved in the nuclear biology of parasites. The prepared polyclonal antibodies against the BgAP2-M peptides further detected a native size of ~ 73 kDa and were localized to the nuclei of B. gibsoni.
    CONCLUSIONS: This study presents a thorough transcriptome analysis of B. gibsoni in vivo and in vitro for the first time, contributing to a detailed understanding of the effects of environmental changes on the growth and development of parasites in the blood stage. Moreover, it also provides a deeper investigation for the different members of the ApiAP2 transcription factor family as various life stage regulators in Babesia spp.
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  • 文章类型: Journal Article
    Plasmodium vivax, the most widely distributed human malaria parasite, is restricted to reticulocytes, limiting its asexual proliferation. In recent years, cases of severe and high-level P. vivax parasitemia have been reported, challenging the assumption that all isolates are equally restricted. In this article, we analyze the reticulocyte preference of a large number of Indian P. vivax isolates. Our results show that P. vivax isolates significantly vary in their level of reticulocyte preference. In addition, by carefully staging the parasites, we find that P. vivax schizonts are largely missing in peripheral blood, with the presence of schizonts in circulation correlating with a high reticulocyte preference.
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