A preliminary investigation of microsatellite-based genotyping in Trichomonas vaginalis.

The genetic epidemiology of Trichomonas vaginalis is poorly understood at present. The recent release of the organism's genome sequence opens the way to investigation of polymorphic markers allowing strain identification. We here report a preliminary analysis of microsatellite loci in T. vaginalis and show that this approach holds promise for future studies of infection transmission and organism diversity.

Bioinformatic analysis of Entamoeba histolytica SINE1 elements.

BACKGROUND: Invasive amoebiasis, caused by infection with the human parasite Entamoeba histolytica remains a major cause of morbidity and mortality in some less-developed countries. Genetically E. histolytica exhibits a number of unusual features including having approximately 20% of its genome comprised of repetitive elements. These include a number of families of SINEs - non-autonomous elements which can, however, move with the help of partner LINEs. In many eukaryotes SINE mobility has had a profound effect on gene expression; in this study we concentrated on one such element - EhSINE1, looking in particular for evidence of recent transposition. RESULTS: EhSINE1s were detected in the newly reassembled E. histolytica genome by searching with a Hidden Markov Model developed to encapsulate the key features of this element; 393 were detected. Examination of their sequences revealed that some had an internal structure showing one to four 26-27 nt repeats. Members of the different classes differ in a number of ways and in particular those with two internal repeats show the properties expected of fairly recently transposed SINEs - they are the most homogeneous in length and sequence, they have the longest (i.e. the least decayed) target site duplications and are the most likely to show evidence (in a cDNA library) of active transcription. Furthermore we were able to identify 15 EhSINE1s (6 pairs and one triplet) which appeared to be identical or very nearly so but inserted into different sites in the genome; these provide good evidence that if mobility has now ceased it has only done so very recently. CONCLUSIONS: Of the many families of repetitive elements present in the genome of E. histolytica we have examined in detail just one - EhSINE1. We have shown that there is evidence for waves of transposition at different points in the past and no evidence that mobility has entirely ceased. There are many aspects of the biology of this parasite which are not understood, in particular why it is pathogenic while the closely related species E. dispar is not, the great genetic diversity found amongst patient isolates and the fact, which may be related, that only a small proportion of those infected develop clinical invasive amoebiasis. Mobile genetic elements, with their ability to alter gene expression may well be important in unravelling these puzzles.

Sexual transmission of intestinal parasites in men who have sex with men.

Direct oral-anal sexual contact is a common practice among men who have sex with men (MSM) and is implicated in the transmission of various enteric pathogens including intestinal parasites. The present study reviewed data on the sexual transmission of intestinal parasites among MSM, and highlighted advances in the diagnosis of such infections. The emergence and spread of intestinal parasites is of public health concern particularly in the homosexual community. Intestinal parasitic infection should be considered in the differential diagnosis of gastrointestinal disease in this population. Combination of traditional diagnostic procedures with implementation of testing based on novel molecular methods in the accurate identification of intestinal parasites is important so that early intervention and control of infection is facilitated.

SINE elements of Entamoeba dispar.

Entamoeba histolytica and E. dispar are closely related protozoan parasites; the former causes clinical amoebiasis in humans while the latter appears to be non-pathogenic. The molecular biology of E. histolytica shows a number of unusual features, one of which is the abundance of polyadenylated but apparently untranslatable mRNAs produced; many of these are the product of at least three families of SINEs (EhSINE1-3). Here we show that the genome of E. dispar contains numerous copies of a SINE element (EdSINE1) whose 5'- and 3'-ends are very similar to those of EhSINE1 but with a much less similar middle portion. Twelve out of 18 copies examined had target site duplications. In none out of six cases examined was there a SINE element in the homologous region of the E. histolytica genome but a single copy of EdSINE1 is present in E. histolytica where it is identified as EhSINE3.

Progress in research on Entamoeba histolytica pathogenesis.

Entamoeba histolytica is a protozoan parasite of humans that causes 40,000-100,000 deaths annually. Clinical amoebiasis results from the spread of the normally luminal parasite into the colon wall and beyond; the key development in understanding this complex multistage process has been the publication of the E. histolytica genome, from which has come an explosion in the use of microarrays to examine changes in gene expression that result from changes in growth conditions. The genome has also revealed a unique arrangement of tRNA genes and an extraordinary number of genes for putative virulence factors, many unexpressed under the artificial conditions of growth in culture. The ability to induce apoptosis of mammalian cells and a useful, but as yet little-understood, technique for epigenetic irreversible gene silencing are other exciting developments.

A bioinformatic analysis of the RAB genes of Trypanosoma brucei.

RAB proteins are small GTPases with vital roles in eukaryotic intracellular transport; orthologous RABs appear to fulfil similar functions in diverse organisms. Trypanosoma brucei spp., the causative organisms of Old World trypanosomiasis of humans and domestic animals, have extremely effective endocytic and exocytic mechanisms that are likely to be involved in maintenance of infection, making study of these systems of importance. Taking advantage of the essential completion of the T. brucei genome, we have re-examined the T. brucei RABs (TbRABs) so far described and identified a total of 16. BLAST searches and phylogenetic analysis show that nine of the TbRABs can confidently be assigned as orthologues or homologues of known RAB proteins from higher eukaryotes, and four more with reasonable probability. The core endocytic pathway is probably similar in complexity to yeast, whilst the early exocytic pathway appears to be more complex than in yeast. Two of the TbRAB family (RAB23 and 28) with clear mammalian orthologues appear to be unusual, and may be involved in nuclear processes and are described in more detail in an accompanying paper. Three TbRABs appear, however, to have no close homologues and may fulfil specialised functions in this organism. The availability of a complete set of TbRABs--which includes orthologues of the RABs responsible for control of the core of the endomembrane system (i.e. RAB1, 2, 4-7 and 11)--provides a first overview of the trafficking complexity that is present within a kinetoplastid parasite. Based on these homologies we suggest a systematic nomenclature for the TbRABs to reflect their functional homologies. This information is of importance both from the perspective of understanding the evolution and diversity of eukaryotic trafficking, but also in providing a framework by which to understand protein processing, trafficking, endocytosis and other related processes in these parasites.

The genome of the protist parasite Entamoeba histolytica.

Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Here we present the genome of E. histolytica, which reveals a variety of metabolic adaptations shared with two other amitochondrial protist pathogens: Giardia lamblia and Trichomonas vaginalis. These adaptations include reduction or elimination of most mitochondrial metabolic pathways and the use of oxidative stress enzymes generally associated with anaerobic prokaryotes. Phylogenomic analysis identifies evidence for lateral gene transfer of bacterial genes into the E. histolytica genome, the effects of which centre on expanding aspects of E. histolytica's metabolic repertoire. The presence of these genes and the potential for novel metabolic pathways in E. histolytica may allow for the development of new chemotherapeutic agents. The genome encodes a large number of novel receptor kinases and contains expansions of a variety of gene families, including those associated with virulence. Additional genome features include an abundance of tandemly repeated transfer-RNA-containing arrays, which may have a structural function in the genome. Analysis of the genome provides new insights into the workings and genome evolution of a major human pathogen.

Nontranslated polyadenylated RNAs from Entamoeba histolytica.

Protozoan parasites display a range of unusual molecular mechanisms that could be helpful for their survival in Nature. Among these parasites, Entamoeba histolytica is one of the most prevalent in developing countries such as India. Entamoeba histolytica produces at least four different unusual transcripts, IE, Tr, ehapt1 and UEE1, that are polyadenylated, but do not have significant open reading frames. Availability of large-scale sequence information has helped us to understand the nature of these sequences and their possible role. Entamoeba histolytica also encodes at least three classes of non-long-terminal-repeats containing retrotransposons, similar to mammalian long retrotransposable elements. This article describes the current status of our understanding of these transcripts and suggests a relationship between some of these transcripts and short retrotransposable element-like retro-elements present in many eukaryotes.

Differentiation of entamoeba histolytica/entamoeba dispar by PCR and their correlation with humoral and cellular immunity in individuals with clinical variants of amoebiasis.

To correlate a particular state of immunity with Entamoeba spp., we used colorimetric PCR to differentiate E. histolytica from E. dispar in individuals with amoebiasis and to associate its presence with the clinical profile, including humoral and cellular immune responses to E. histolytica. Our results showed high levels of antibody in acute amoebiasis and elevation of IL-4 production, a cytokine related to Th2 profile, associated with E. histolytica. In chronic amoebiasis, even with anti-E. histolytica seropositivity, intestinal symptoms were associated with E. dispar in all the cases, without differences in level of antibodies, BTI, CD4+/CD8+ ratio, INF-gamma, and IL-4. Among asymptomatic carriers, E. dispar was more frequently found; however, identification of E. histolytica in two asymptomatic carriers associated with high levels of INF-gamma, a cytokine related to Th1 profile, demonstrate the importance of making specific diagnosis of Entamoeba spp., to establish the clinical and epidemiological behavior in both intestinal and extra-intestinal amoebiasis.