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      Characterization of a novel Entamoeba histolytica strain from Burkina Faso, Africa, possessing a unique hexokinase-2 gene Translated title: Caractérisation d’une nouvelle souche d’ Entamoeba histolytica au Burkina Faso, Afrique, présentant un gène unique codant pour l’hexokinase-2

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          Abstract

          An Entamoeba histolytica strain (BF-841 cl1) that originated from Burkina Faso, Africa presented with novel, polymorphic genotypes of the serine-rich E. histolytica protein and the anodic hexokinase-2 (HXK-2) isoenzyme band, which showed less electrophoretic mobility than that of an E. histolytica reference strain [HM-1:IMSS cl6 (zymodeme (Z)-II)] by starch gel electrophoresis and isoelectric focusing (IEF). The HXK-2 gene of BF-841 cl1 had amino acid variations at four positions compared to the sequence of HM-1: IMSS cl6. These variations were absent from the sequences of four other E. histolytica strains with different zymodemes [KU27 (Z-II), SAW1627 (Z-IIα-), SAW755CR clB (Z-XIV), and KU2 (Z-XIX)]. The results of IEF showed no difference in the substrate specificity of HXK (HXK-1 and HXK-2) between BF-841 cl1 and the three reference E. histolytica strains (HM-1:IMSS cl6, SAW755 clB, and KU27). It was also confirmed that BF-841 cl1 was able to form liver abscesses in Syrian hamsters.

          Translated abstract

          Une souche d’ Entamoeba histolytica (BF-841 cl1), originaire du Burkina Faso en Afrique, présente un nouveau polymorphisme génétique pour la protéine riche en sérine et aussi pour l’hexokinase anodine (HXK-2) qui a une moindre mobilité électrophorétique que celle de la souche d’ E. histolytica de référence [HM-1:IMSS cl6 (zymodème (Z)-II)] en électrophorèse sur gel d’amidon et dans l’analyse en focalisation isoélectrique (IEF). Le gène HXK-2 de la souche BF-841 cl1 présente des variations d’acides aminés à quatre positions par rapport à la souche de HM-I:IMSS cl6. Ces variations diffèrent aussi de celles de quatre autres souches d’ E. histolytica avec différents zymodèmes [KU27 (Z-II), SAW1627 (Z-IIα-), SAW755CR clB (Z-XIV) et KU2 (Z-XIX)]. Les résultats de l’IEF ne montrent aucune différence dans la spécificité du substrat de HXK (HXK-1 et HXK-2) entre la souche BF-841 cl1 et les trois souches référencées (HM-1:IMSS cl6, SAW755 clB et KU27). Il a aussi été confirmé que BF-841 cl1 est capable de former un abcès du foie chez des hamsters syriens.

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          Most cited references 16

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          The SWISS-MODEL Repository and associated resources

          SWISS-MODEL Repository (http://swissmodel.expasy.org/repository/) is a database of 3D protein structure models generated by the SWISS-MODEL homology-modelling pipeline. The aim of the SWISS-MODEL Repository is to provide access to an up-to-date collection of annotated 3D protein models generated by automated homology modelling for all sequences in Swiss-Prot and for relevant models organisms. Regular updates ensure that target coverage is complete, that models are built using the most recent sequence and template structure databases, and that improvements in the underlying modelling pipeline are fully utilised. As of September 2008, the database contains 3.4 million entries for 2.7 million different protein sequences from the UniProt database. SWISS-MODEL Repository allows the users to assess the quality of the models in the database, search for alternative template structures, and to build models interactively via SWISS-MODEL Workspace (http://swissmodel.expasy.org/workspace/). Annotation of models with functional information and cross-linking with other databases such as the Protein Model Portal (http://www.proteinmodelportal.org) of the PSI Structural Genomics Knowledge Base facilitates the navigation between protein sequence and structure resources.
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            Remarkable genetic polymorphism among Entamoeba histolytica isolates from a limited geographic area.

            In order to understand genetic polymorphisms among Entamoeba histolytica strains in a limited geographic area and among restricted social populations, we studied nucleotide polymorphism in DNA regions that do not encode proteins (locus 1-2 and locus 5-6) and in genes coding for chitinase and for serine-rich E. histolytica protein. Thirty E. histolytica isolates from domestically infected Japanese amebiasis patients (male homosexuals and residents in institutions for the mentally handicapped) and four reference strains were examined. PCR revealed remarkable polymorphisms in both the number and size of the PCR fragments containing these loci. Polymorphisms in lengths, types, and numbers of internal repeat units were observed in locus 1-2 and the repeat-containing region of serine-rich E. histolytica protein among the Japanese isolates. In contrast, polymorphism at locus 5-6 was observed almost exclusively in the number of repeats of a 16-nucleotide unit. The repeat-containing region of chitinase appeared to be the least polymorphic among the four loci with a single dominant genotype representing 66% (20 out of 30) of all of the isolates. Isolates obtained from male homosexuals showed a more complex genetic polymorphism than those from residents in institutions. Considering all four polymorphic loci together, all 19 Japanese isolates from male homosexuals were distinct. In contrast, all isolates obtained from mass-infection cases at a single institution had an identical genotype, suggesting that these cases were caused by a single E. histolytica strain. No significant correlation was found between genotypes and zymodemes or between genotypes and clinical presentations, e.g., colitis or liver abscess. Certain genotypes were observed with higher frequencies in male homosexuals or residents of institutions. These data indicate that genotyping of the E. histolytica isolates by using these four polymorphic loci could serve as a tool to fingerprint individual isolates. We propose that genotyping of ameba isolates should help to determine geographic origins of isolates and routes of transmission.
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              Geographic diversity among genotypes of Entamoeba histolytica field isolates.

              It has been known that only 5 to 10% of those infected with Entamoeba histolytica develop symptomatic disease. However, the parasite and the host factors that determine the onset of disease remain undetermined. Molecular typing by using polymorphic genetic loci has been proven to aid in the close examination of the population structure of E. histolytica field isolates in nature. In the present study, we analyzed the genetic polymorphisms of two noncoding loci (locus 1-2 and locus 5-6) and two protein-coding loci (chitinase and serine-rich E. histolytica protein [SREHP]) among 79 isolates obtained from different geographic regions, mainly Japan, Thailand, and Bangladesh. When the genotypes of the four loci were combined for all isolates that we have analyzed so far (overlapping isolates from mass infection events were excluded), a total of 53 different genotypes were observed among 63 isolates. The most remarkable and extensive variations among the four loci was found in the SREHP locus; i.e., 34 different genotypes were observed among 52 isolates. These results demonstrate that E. histolytica has an extremely complex genetic structure independent of geographic location. Our results also show that, despite the proposed transmission of other sexually transmitted diseases, including human immunodeficiency virus infection, from Thailand to Japan, the spectra of the genotypes of the E. histolytica isolates from these two countries are distinct, suggesting that the major E. histolytica strains prevalent in Japan at present were likely introduced from countries other than Thailand. Although the genetic polymorphism of the SREHP locus was previously suggested to be closely associated with the clinical presentation, e.g., colitis or dysentery and liver abscess, no association between the clinical presentation and the SREHP genotype at either the nucleotide or the predicted amino acid level was demonstrated.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                November 2011
                15 November 2011
                : 18
                : 4 ( publisher-idID: parasite/2011/04 )
                : 287-294
                Affiliations
                [1 ] Division of Clinical Microbiology, Department of Microbiology, Tokyo Metropolitan Institute of Public Health Tokyo Japan
                [2 ] Department of Tropical Medicine and Parasitology, School of Medicine, Keio University Tokyo Japan
                [3 ] Department of Medical Genome Science, Graduate School of Frontier Sciences, University of Tokyo Tokyo Japan
                Author notes
                [* ]Correspondence: Seiki Kobayashi, Ph.D., Department of Tropical Medicine and Parasitology, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan. Tel.: 81 3 5363 3761 – Fax: 81 3 3353 5958. E-mail: skobaya@ 123456sc.itc.keio.ac.jp
                Article
                parasite2011184p287 10.1051/parasite/2011184287
                10.1051/parasite/2011184287
                3677594
                22091458
                © PRINCEPS Editions, Paris, 2011

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 3, Tables: 4, Equations: 1, References: 21, Pages: 8
                Categories
                Original Contribution

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