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1.
BMC Infect Dis ; 20(1): 906, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-33256616

RESUMO

BACKGROUNDS: Primary infection with Toxoplasma gondii during pregnancy can pose serious health problems for the fetus. However, the epidemiological status of toxoplasmosis among reproductive-aged population in Myanmar is largely unknown. Although luciferase immunoprecipitation system (LIPS) assays for serodiagnosis of toxoplasmosis was developed mostly using mouse infection model, had not been tested by using field-derived human samples. METHODS: A total of 251 serum samples were collected from reproductive-aged women, residing in Shwegyin township, Bago region, Myanmar and analyzed with a commercial ELISA kit, as well as in-house LIPS assays. RESULTS: The overall seroprevalence for Toxoplasma gondii infection by the commercial ELISA was 11.5%. No clear risk factor was identified except for being in the younger age group (15-30 years old). Overall, LIPS assays showed low sensitivity when the commercial ELSA was used as a reference test. CONCLUSION: We identified the epidemiological situation of toxoplasmosis in some rural communities in Myanmar. The data obtained here will serve as a primary information for the effort to reduce toxoplasmosis in this region. Although looked promising in the previous experiments with mouse infection model, we found that the reported LIPS procedures need further improvements to increase the sensitivities.


Assuntos
Imunoprecipitação/métodos , Medições Luminescentes/métodos , Testes Sorológicos/métodos , Toxoplasma/imunologia , Toxoplasmose/diagnóstico , Toxoplasmose/epidemiologia , Adolescente , Adulto , Animais , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Luciferases , Substâncias Luminescentes , Camundongos , Pessoa de Meia-Idade , Mianmar/epidemiologia , Fatores de Risco , População Rural , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Toxoplasmose/sangue , Toxoplasmose/parasitologia , Adulto Jovem
3.
Genome Res ; 24(8): 1384-95, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24755901

RESUMO

Although many de novo genome assembly projects have recently been conducted using high-throughput sequencers, assembling highly heterozygous diploid genomes is a substantial challenge due to the increased complexity of the de Bruijn graph structure predominantly used. To address the increasing demand for sequencing of nonmodel and/or wild-type samples, in most cases inbred lines or fosmid-based hierarchical sequencing methods are used to overcome such problems. However, these methods are costly and time consuming, forfeiting the advantages of massive parallel sequencing. Here, we describe a novel de novo assembler, Platanus, that can effectively manage high-throughput data from heterozygous samples. Platanus assembles DNA fragments (reads) into contigs by constructing de Bruijn graphs with automatically optimized k-mer sizes followed by the scaffolding of contigs based on paired-end information. The complicated graph structures that result from the heterozygosity are simplified during not only the contig assembly step but also the scaffolding step. We evaluated the assembly results on eukaryotic samples with various levels of heterozygosity. Compared with other assemblers, Platanus yields assembly results that have a larger scaffold NG50 length without any accompanying loss of accuracy in both simulated and real data. In addition, Platanus recorded the largest scaffold NG50 values for two of the three low-heterozygosity species used in the de novo assembly contest, Assemblathon 2. Platanus therefore provides a novel and efficient approach for the assembly of gigabase-sized highly heterozygous genomes and is an attractive alternative to the existing assemblers designed for genomes of lower heterozygosity.


Assuntos
Mapeamento de Sequências Contíguas , Software , Animais , Caenorhabditis elegans/genética , Genoma Helmíntico , Heterozigoto , Ostreidae/genética , Análise de Sequência de DNA
4.
Eukaryot Cell ; 12(12): 1588-99, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23873863

RESUMO

Microtubules are versatile biopolymers that support numerous vital cellular functions in eukaryotes. The specific properties of microtubules are dependent on distinct microtubule-associated proteins, as the tubulin subunits and microtubule structure are exceptionally conserved. Highly specialized microtubule-containing assemblies are often found in protists, which are rich sources for novel microtubule-associated proteins. A protozoan parasite, Toxoplasma gondii, possesses several distinct tubulin-containing structures, including 22 microtubules closely associated with the cortical membrane. Early ultrastructural studies have shown that the cortical microtubules are heavily decorated with associating proteins. However, little is known about the identities of these proteins. Here, we report the discovery of a novel protein, TrxL1 (for Thioredoxin-Like protein 1), and an associating complex that coats the cortical microtubules. TrxL1 contains a thioredoxin-like fold. To visualize its localization in live parasites by fluorescence, we replaced the endogenous TrxL1 gene with an mEmeraldFP-TrxL1 fusion gene. Structured illumination-based superresolution imaging of this parasite line produced a detailed view of the microtubule cytoskeleton. Despite its stable association with the cortical microtubules in the parasite, TrxL1 does not seem to bind to microtubules directly. Coimmunoprecipitation experiments showed that TrxL1 associates with a protein complex containing SPM1, a previously reported microtubule-associated protein in T. gondii. We also found that SPM1 recruits TrxL1 to the cortical microtubules. Besides SPM1, several other novel proteins are found in the TrxL1-containing complex, including TrxL2, a close homolog of TrxL1. Thus, our results reveal for the first time a microtubule-associated complex in T. gondii.


Assuntos
Microtúbulos/metabolismo , Proteínas de Protozoários/metabolismo , Tiorredoxinas/metabolismo , Toxoplasma/metabolismo , Toxoplasmose/parasitologia , Sequência de Aminoácidos , Humanos , Microtúbulos/química , Microtúbulos/genética , Dados de Sequência Molecular , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas de Protozoários/química , Proteínas de Protozoários/genética , Alinhamento de Sequência , Tiorredoxinas/química , Tiorredoxinas/genética , Toxoplasma/química , Toxoplasma/genética
5.
Clin J Gastroenterol ; 17(1): 143-147, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38085443

RESUMO

Anisakiasis is a parasitic infection caused by the ingestion of raw or undercooked seafood infected with Anisakis larvae. It generally affects the gastrointestinal tract, particularly the stomach, but very rare cases have been reported in which infection of the liver leads to the formation of inflammatory pseudotumors. We herein report an extremely rare case of an inflammatory pseudotumor induced by hepatic anisakiasis that was laparoscopically resected for the purpose of both diagnosis and treatment. A 51-year-old woman underwent a routine medical checkup by ultrasound examination, which incidentally detected a 15-mm mass on the surface of S6 of the liver. Because a malignant tumor could not be ruled out on several preoperative imaging studies, laparoscopic partial resection of the liver was performed. Histopathological examination revealed Anisakis larva in the inflammatory pseudotumor, suggesting hepatic anisakiasis. This report describes an extremely rare case of an inflammatory pseudotumor induced by hepatic anisakiasis. Because the preoperative diagnosis could not be obtained by several imaging modalities, laparoscopic liver resection with a sufficient margin might be suitable for diagnosis and treatment of this disease.


Assuntos
Anisaquíase , Anisakis , Granuloma de Células Plasmáticas , Animais , Feminino , Humanos , Pessoa de Meia-Idade , Anisaquíase/diagnóstico , Anisaquíase/cirurgia , Anisaquíase/parasitologia , Granuloma de Células Plasmáticas/diagnóstico por imagem , Granuloma de Células Plasmáticas/cirurgia , Estômago/patologia , Larva , Fígado/cirurgia , Fígado/patologia
6.
Parasitol Int ; 92: 102663, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36058466

RESUMO

Strongyloides is a genus of parasitic nematodes of vertebrates comprising approximately 50 documented species, each with various host ranges. Among these, three species (S. stercoralis, S. fuelleborni, and S. cebus) are known to infect primate hosts. S. fuelleborni typically infects non-human primates in the Old World. To complement the existing information on the global genetic structure of this species, we conducted a genotyping study of S. fuelleborni samples collected from rhesus macaques in Myanmar, Japanese macaques in Japan, and some zoo-kept primates. This study identified a novel haplotype group in isolates from the Myanmar rhesus macaques. Subsequently, we obtained the complete or nearly complete mitochondrial genome sequences of S. fuelleborni, S. cebus (Strongyloides of New World monkeys), and S. vituli (Strongyloides of cattle). Phylogenetic analysis based on concatenated mitochondrial protein sequences of various Strongyloides species indicated a close relationship between S. fuelleborni, S. vituli and S. papillosus (Strongyloides in sheep and cattle). S. cebus is quite distantly related to both S. fuelleborni and S. stercoralis, which led to the hypothesis that the three primate Strongyloides species evolved independently as parasites of primates.


Assuntos
Genoma Mitocondrial , Bovinos , Ovinos , Animais , Filogenia , Strongyloides/genética , Macaca mulatta , Cebus , Macaca fuscata , Genética Populacional
7.
Parasitol Res ; 110(4): 1583-6, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22006188

RESUMO

C-type lectins (CTLs) are a group of proteins which bind to carbohydrate epitopes in the presence of Ca(2+), which have been described in a wide range of species. In this study, a cDNA sequence coding a putative CTL has been identified from the cDNA library constructed from the pig round worm Ascaris suum lung L3 (LL3) larvae, which was designated as A. suum C-type lectin-1 (As-CTL-1). The 510 nucleotide open reading frame of As-CTL-1 cDNA encoded the predicted 169 amino acid protein including a putative signal peptide of 23 residues and C-type lectin/C-type lectin-like domain (CLECT) at residue 26 to 167. As-CTL-1 was most similar to Toxocara canis C-type lectin-1 and 4 (Tc-CTL-1 and 4), and highly homologous to namatode CTLs and mammalian CTLs as well, such as human C-type lectin domain family 4 member G (CLECG4). In addition, As-CTL-1 was strongly expressed in tissue migrating LL3 and the L4 larvae, which were developmental larvae stages within the mammalian host. These results suggest that A. suum larvae might utilize As-CTL-1 to avoid pathogen recognition mechanisms in mammalian hosts due to it is similarity to host immune cell receptors.


Assuntos
Ascaris suum/química , Etiquetas de Sequências Expressas , Larva/metabolismo , Lectinas Tipo C/isolamento & purificação , Animais , Ascaris suum/genética , Clonagem Molecular , Biblioteca Gênica , Lectinas Tipo C/química , Lectinas Tipo C/genética , Masculino , RNA Ribossômico 18S/genética , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência , Análise de Sequência de DNA/métodos , Toxocara canis/química
8.
J Vet Med Sci ; 84(7): 905-913, 2022 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-35584943

RESUMO

Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among serological diagnostic methods, Luciferase-linked Antibody Capture Assay (LACA) has been found to be a promising platform with high sensitivity and specificity. In this study, we aimed to evaluate recombinant nanoluciferase fused-T. gondii antigens (rNluc-GRA6, rNluc-GRA7, rNluc-GRA8 and rNluc-BAG1) for their potential use in LACA for pigs. As a result, the sensitivity of GRA6-, GRA7-, GRA8- and BAG1-LACA were 70.0%, 80.0%, 80.0% and 30.0% with specificity 87.0%, 81.5%, 74.1% and 50.0%, respectively. The cocktail LACA using a mixture of rNluc-GRA6, rNluc-GRA7 and rNluc-GRA8 indicated higher sensitivity (90.0%) and a similar specificity (96.3%) in comparison with the commercial ELISA kit. Compared to the Dye-Test as a reference test, cocktail LACA showed strong agreement (kappa value=0.811) when we assessed pig sera collected at the slaughterhouse. In addition, we also successfully established the rapid LACA format for the detection of Toxoplasma infection in pigs (called Rapid-LACA) in which the test could be performed within 30 min. In Rapid-LACA, the protein A pre-coated/blocked plates could be preserved at -30°C, 4°C or room temperature conditions for at least two months without compromising on the quality of assay.


Assuntos
Doenças dos Suínos , Toxoplasma , Toxoplasmose Animal , Toxoplasmose , Animais , Anticorpos Antiprotozoários , Antígenos de Protozoários , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/veterinária , Luciferases/genética , Luciferases/metabolismo , Proteínas de Protozoários , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/diagnóstico , Toxoplasmose Animal/diagnóstico
9.
Parasitol Int ; 84: 102399, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34077794

RESUMO

Strongyloides is a genus of parasitic nematodes of vertebrates that contains over 50 species, each with a variable host range. A recent molecular phylogenetic analysis on this genus showed that Strongyloides spp. from various carnivore hosts form a strongly supported clade together with Strongyloides stercoralis, a major pathogen of humans and dogs (named the "stercoralis/procyonis group"). In the present study, we obtained DNA sequencing data of Strongyloides sp. isolated from an imported meerkat (Suricata suricatta). Based on the phylogenetic analysis, we considered this a new member of the stercoralis/procyonis group. This study represents the first isolation and molecular characterization of a Strongyloides species from hosts belonging to the family Herpestidae (mongooses and meerkat). However, whether the meerkat serves as a natural host of this Strongyloides species remains to be investigated.


Assuntos
Herpestidae , Strongyloides/classificação , Estrongiloidíase/veterinária , Animais , Sequência de Bases , DNA de Helmintos/análise , Masculino , Animais de Estimação , Strongyloides/genética , Strongyloides/isolamento & purificação , Estrongiloidíase/diagnóstico , Estrongiloidíase/parasitologia
10.
Mod Rheumatol Case Rep ; 5(1): 16-21, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32772699

RESUMO

An elderly woman with rheumatoid arthritis (RA) presented with a chief complaint of abdominal pain and diarrhoea while undergoing treatment with low-dose corticosteroids and abatacept. Endoscopic and histopathological findings revealed manifestations of ulcerative colitis (UC). An intermediate dose of corticosteroids and 5-aminosalicylic acid were administered. Abatacept was discontinued; the anti-TNF biologic, golimumab, was administered for treatment of both RA and UC. However, colitis worsened in response to this therapeutic regimen. Colonoscopy revealed severe mucosal lesions; larvae were detected in samples taken from multiple shallow mucosal ulcers. The patient was diagnosed with Strongyloides stercoralis colitis based on the results of an anti-parasite antibody test and examination of the larval DNA. Furthermore, serology revealed a positive test for antibodies against human T-cell leukaemia virus type 1 (HTLV-1). Immunosuppressive treatment was terminated; ivermectin was administered, which resulted in improvements in colitis symptoms within a few weeks. There are several published reports describing S. stercoralis colitis as a lethal mimic of UC. Corticosteroid and anti-TNF therapies have been reported as among the major risk factors associated with strongyloidiasis in patients with HTLV-1 infection. Therefore, HTLV-1 and Strongyloides infections may be considered in cases of new-onset gastrointestinal symptoms during immunosuppressive therapy, particularly in HTLV-1-endemic regions.


Assuntos
Artrite Reumatoide/complicações , Colite Ulcerativa/induzido quimicamente , Infecções por HTLV-I/complicações , Imunossupressores/efeitos adversos , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/complicações , Idoso , Animais , Antiparasitários/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Colite Ulcerativa/patologia , Feminino , Infecções por HTLV-I/diagnóstico , Infecções por HTLV-I/tratamento farmacológico , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Humanos , Ivermectina/uso terapêutico , Estrongiloidíase/diagnóstico , Estrongiloidíase/tratamento farmacológico , Inibidores do Fator de Necrose Tumoral/uso terapêutico
11.
Parasitol Int ; 84: 102405, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34139361

RESUMO

Ancylostoma ceylanicum is recognized as the only zoonotic hookworm species that is able to mature into adult stage in the human intestine. While human infections caused by this hookworm species have been reported from neighboring countries and this hookworm is prevalent in dogs in Vietnam, human infection has never been reported in Vietnam. The present study, therefore, aimed to identify human infections with A. ceylanicum in Vietnam. A total of 526 fecal samples from the residents in Long An Province were collected and the presence of hookworm eggs was detected by the Kato-Katz method. The results indicated that the overall prevalence of human hookworm infection was 85/526 (16.2%). After filter paper culture, 3rd stage larvae were successfully obtained from 48 egg-positive samples. The larvae were identified for their species using semi-nested PCR-RLFP on the cox1 gene. As a result, two hookworm species were confirmed; single species infections with Necator americanus or A. ceylanicum, and mixed infections with both species were found in 47.9%, 31.3%, and 20.8% of the samples, respectively.


Assuntos
Ancylostoma/isolamento & purificação , Ancilostomíase/epidemiologia , Ancilostomíase/parasitologia , Animais , Complexo IV da Cadeia de Transporte de Elétrons/análise , Proteínas de Helminto/análise , Humanos , Prevalência , Vietnã/epidemiologia
12.
Parasitol Int ; 77: 102125, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32311471

RESUMO

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes the most common parasitic zoonosis worldwide in multiples species of mammals and birds. Although free-range chickens may play a role as an important reservoir for T. gondii, there is no reliable and commercially available diagnostic test for this disease in chickens. In this study, we aimed to develop a novel Luciferase-linked Antibody Capture Assay (LACA) for the serodiagnosis of Toxoplasma infection in chickens. Recombinant nanoluciferase fused-T. gondii dense granule antigen 8 (rNluc-GRA8) was produced and applied to LACA assay as a diagnostic antigen. GRA8-LACA was tested with the sera from uninfected and experimentally infected chickens with T. gondii and other parasitic pathogens and showed unexpectedly high sensitivity (90.5%) and specificity (95.4%). Interestingly, E. coli lysate expressing rNluc-GRA8 could be applied in GRA8-LACA with 85.7% sensitivity and an increased specificity (96.9%) that gave better diagnostic performance compared to conventional ELISA. We applied our diagnostic system to examine 267 free-range chicken sera collected from 12 farms and 100 closed-house broiler chicken sera from local poultry abattoirs. The overall seroprevalence of toxoplasmosis in free-range chickens was 10.9% (95% CI: 10.6%-11.1%), while no positive case was found in broiler chickens. GRA8-LACA could be a useful diagnostic technique for T. gondii infection in chickens. The detection of T. gondii seropositive chickens in this study warns a potential risk of Toxoplasma transmission by the consumption of raw or undercooked chicken meat.


Assuntos
Anticorpos Antiprotozoários/análise , Reservatórios de Doenças/veterinária , Doenças das Aves Domésticas/diagnóstico , Testes Sorológicos/métodos , Toxoplasmose Animal/diagnóstico , Animais , Antígenos de Protozoários/imunologia , Galinhas/parasitologia , Reservatórios de Doenças/parasitologia , Luciferases/química , Masculino , Aves Domésticas/parasitologia , Doenças das Aves Domésticas/parasitologia , Proteínas de Protozoários/imunologia , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Toxoplasma/imunologia , Toxoplasmose Animal/imunologia
13.
BMC Mol Cell Biol ; 21(1): 8, 2020 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-32111164

RESUMO

BACKGROUND: TgDCX is a doublecortin-domain protein associated with the conoid fibers, a set of strongly curved non-tubular tubulin-polymers in Toxoplasma. TgDCX deletion impairs conoid structure and parasite invasion. TgDCX contains two tubulin-binding domains: a partial P25α and the DCX/doublecortin domain. Orthologues are found in apicomplexans and their free-living relatives Chromera and Vitrella. RESULTS: We report that isolated TgDCX-containing conoid fibers retain their pronounced curvature, but loss of TgDCX destabilizes the fibers. We crystallized and determined the 3D-structure of the DCX-domain, which is similar to those of human doublecortin and well-conserved among TgDCX orthologues. However, the orthologues vary widely in targeting to the conoid in Toxoplasma and in modulating microtubule organization in Xenopus cells. Several orthologues bind to microtubules in Xenopus cells, but only TgDCX generates short, strongly curved microtubule arcs. EM analysis shows microtubules decorated with TgDCX bundled into rafts, often bordered on one edge by a "C"-shaped incomplete tube. A Chromera orthologue closely mimics TgDCX targeting in Toxoplasma and binds to microtubules in Xenopus cells, but does not generate arcs or "C"-shaped tubes, and fails to rescue the defects of the TgDCX-knockout parasite. CONCLUSIONS: These observations suggest that species-specific features of TgDCX enable it to generate strongly curved tubulin-polymers to support efficient host-cell invasion.


Assuntos
Proteínas Associadas aos Microtúbulos/química , Neuropeptídeos/química , Toxoplasma/metabolismo , Tubulina (Proteína)/metabolismo , Animais , Células Cultivadas , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Proteínas do Domínio Duplacortina , Proteína Duplacortina , Técnicas de Inativação de Genes , Interações Hospedeiro-Parasita/genética , Microscopia Eletrônica , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Associadas aos Microtúbulos/ultraestrutura , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Microtúbulos/ultraestrutura , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Polímeros/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Domínios Proteicos/genética , Proteínas Recombinantes , Toxoplasma/química , Toxoplasma/efeitos dos fármacos , Toxoplasma/ultraestrutura , Tubulina (Proteína)/química , Xenopus
14.
Parasitol Int ; 78: 102151, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32502520

RESUMO

Strongyloides stercoralis is a parasitic nematode and a major pathogen responsible for human strongyloidiasis. The presence of this species in the dog population has led to an interest in studying the phylogenetic relationships among Strongyloides spp. in carnivore hosts. In the present study, Strongyloides spp. from various carnivore hosts (raccoon, Japanese badger, Siberian weasel, raccoon dog, masked palm civet, and domestic cat) were sought. Except for civets, Strongyloides spp. were identified in all host species. Based on 18S rDNA sequences, nine OTUs (operational taxonomy units) were identified. Molecular phylogenetic analyses using 18S28S rDNA and mitochondrial cox1 (cytochrome c oxidase subunit 1) sequences clustered them into two groups. The first group (named the stercoralis/procyonis group) was comprised of six OTUs and occurred in cats, raccoon dogs, raccoons (S. procyonis), Siberian weasels, and Japanese badgers and included S. stercoralis from humans and dogs. The second group (named the planiceps group) was made up of Strongyloides spp. from raccoon dogs (two OTUs) and one OTU from Siberian weasels. Subsequent analysis using almost the full-length nucleotide sequences of protein-coding genes in their mitochondrial genomes placed Strongyloides spp. of cats in a sister taxon position to S. stercoralis, whereas S. procyonis from raccoons was more distantly related to them. The presence of Strongyloides spp. from various carnivore hosts, which are close relatives of S. stercoralis, suggests this group of Strongyloides (the stercoralis/procyonis group) essentially evolved as parasites of carnivores, although more data on Strongyloides spp. from primate hosts are needed.


Assuntos
Carnívoros , Strongyloides/classificação , Estrongiloidíase/veterinária , Animais , Feminino , Japão , Mianmar , Filogenia , RNA de Helmintos/análise , RNA Ribossômico 18S/análise , Strongyloides/fisiologia , Estrongiloidíase/parasitologia
15.
J Microbiol Immunol Infect ; 52(1): 100-105, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28754237

RESUMO

BACKGROUND: Larva migrans syndrome is a food-borne parasitic disease in humans, caused by accidental ingestion of eggs or larvae of ascarid nematodes, namely, Toxocara canis, Toxocara cati, or Ascaris suum, the roundworms commonly found in the intestines of dogs, cats and pigs respectively. When a patient is diagnosed as having larva migrans syndrome, oral-administration of albendazole is recommended, however, the regimen remains controversial worldwide. In Japan, the duration of albendazole administration is longer than those of European and North American countries. The purpose of this study was to assess the efficacy and safety of long-term administration treatment of albendazole for larva migrans syndrome. METHODS: From 2004 to 2014, our laboratory was involved in the diagnosis of 758 larva migrans syndrome cases, of which 299 cases could be followed up after the treatment. We analyzed these 299 follow-up cases on the ELISA results before and after the treatment as well as on anthelmintic used, dose and duration of medication, clinical findings, and side effects, recorded on a consultation sheet provided by the attending physicians. We have 288 cases as the subjects of this study. RESULTS: Albendazole represented a 78.0% efficacy rate. The side effects represented 15.0% in using albendazole alone cases; however, the side effects were mild to moderate and there were no severe cases reported. CONCLUSIONS: The long-term administration treatment of albendazole is safe and effective for larva migrans syndrome.


Assuntos
Albendazol/uso terapêutico , Anti-Helmínticos/uso terapêutico , Larva Migrans/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Albendazol/efeitos adversos , Animais , Anti-Helmínticos/efeitos adversos , Anticorpos Anti-Helmínticos/análise , Ascaridoidea/imunologia , Criança , Feminino , Seguimentos , Humanos , Japão , Larva/imunologia , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Adulto Jovem
16.
Parasitol Int ; 72: 101937, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31158501

RESUMO

Traditionally, Strongyloides nematode infecting cattle had been thought to be a single species, S. papillosus. Surprisingly, Eberhardt et al. in 2008 reported two, rather than one Strongyloides species infected cattle, with one being S. papillosus and the other S. vituli. However, there was no subsequent report to support their findings. In July 2018, a case of a sudden death of a calf believed to be due to heavy infection with S. papillosus at a dairy farm in Miyazaki Prefecture, Japan, was reported. One month after the initiation of a deworming program to prevent further sudden deaths, fecal specimens from 24 cattle housed in the same barn were examined. Eight samples were positive for Strongyloides eggs. For species determination, the nucleotide sequences of 18S rDNA (small subunit ribosomal DNA gene), rpl-10 (ribosomal protein L10 gene), and mitochondrial (mt) cox1 (cytochrome c oxidase subunit 1 gene) were obtained. Typing data for all three marker genes indicated the presence of both species, S. papillosus and S. vituli, in the fecal samples. To our knowledge, this study is the first to support the original report by Eberhardt et al. regarding the sympatric existence of S. papillosus and S. vituli in cattle, and to report the presence of S. vituli in Japan. Interestingly, phylogenetic analyses of both rpl-10 and mt cox1 sequences indicated a closer genetic relationship of S. vituli with S. venezuelensis (Strongyloides of rats) than with S. papillosus, shedding light on the speciation history of Strongyloides nematodes.


Assuntos
Doenças dos Bovinos/epidemiologia , Bovinos/parasitologia , Evolução Molecular , Strongyloides/genética , Estrongiloidíase/veterinária , Animais , Doenças dos Bovinos/parasitologia , Ciclo-Oxigenase 1/genética , DNA Ribossômico/genética , Fezes/parasitologia , Feminino , Japão , Filogenia , Strongyloides/isolamento & purificação , Estrongiloidíase/epidemiologia
17.
J Immunol Methods ; 462: 91-100, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30201391

RESUMO

Development of reliable, quantitative technologies for serodiagnosis of Toxoplasma gondii infection remains desirable. The luciferase immunoprecipitation system (LIPS) is a relatively simple, highly sensitive, and rapid quantitative immunoassay. The major advantages of this assay over ELISA are a wider dynamic range, shorter overall assay time, and less sample volume. In this study, we aimed to use this method for the serodiagnosis of toxoplasmosis. Recombinant Toxoplasma antigens (dense granule antigens GRA6, GRA7, and GRA8 and bradyzoite antigen BAG1) fused with nanoluciferase (Nluc, a small luciferase enzyme) were expressed in Escherichia coli, purified, and tested in LIPS assays with sera from experimental mice infected with T. gondii and a WHO standard anti-Toxoplasma human immunoglobulin (TOXM). In the experimentally infected mice, LIPS assays detected antibodies against Nluc-GRA6, Nluc-GRA7, and Nluc-GRA8 as early as day 14, whereas antibodies against Nluc-BAG1 remained undetected until day 21 and then showed significant elevation on day 60. In TOXM sera, LIPS assays with each Nluc recombinant protein produced reliable standard curves with a coefficient of determination (R2) of 0.980-0.989 for GRA6, 0.986-0.990 for GRA7, 0.998-0.999 for GRA8, and 0.942-0.987 for BAG1. The detection limits were estimated to be 3.9, 2, 1, and 1 IU/ml for rGRA6, rGRA7, rGRA8, and rBAG1, respectively. The LIPS assay for toxoplasmosis could detect antibodies against T. gondii in the mouse and human sera with a reasonably high sensitivity. We consider the LIPS assay to be a promising alternative tool for screening, diagnosing, and monitoring toxoplasmosis. In particular, detection of antibodies against BAG1 may be useful for a longitudinal seroprevalence study in suspected high-risk areas on the basis of its elevated serum concentration in the chronic phase.


Assuntos
Anticorpos Antiprotozoários , Antígenos de Protozoários , Imunoprecipitação/métodos , Luciferases , Testes Sorológicos/métodos , Toxoplasma , Toxoplasmose , Animais , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/sangue , Antígenos de Protozoários/imunologia , Feminino , Humanos , Luciferases/imunologia , Luciferases/metabolismo , Camundongos , Toxoplasma/imunologia , Toxoplasma/metabolismo , Toxoplasmose/sangue , Toxoplasmose/diagnóstico , Toxoplasmose/imunologia
18.
Trop Med Health ; 46: 43, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30598622

RESUMO

BACKGROUND: Strongyloidiasis is prevalent in Southeast Asian regions along with other soil-transmitted helminthiases, but only limited present-day data was available for Myanmar. METHODS: A prevalence survey for Strongyloides stercoralis infection was conducted among villagers in rural areas of three townships located in the Lower Myanmar during 2014-2016 by agar plate culture method in combination with specific identification by molecular assays. Risk factors associated with S. stercoralis infection were assessed by analyzing questionnaires obtained from study participants. RESULTS: Strongyloides stercoralis was identified in 40 out of 703 participants (5.7% overall prevalence). The highest prevalence (14.4%) was observed in Htantabin, while other two communities (Thabaung and Thanlyin) had much lower prevalence (2.2 and 2.5%, respectively). Infection was relatively rare (1.2%) in younger generations under 20 years compared to older generations (9.5%). Even in Htantabin, none of the female residents under age 40 (n = 33) had infection. In adult Htantabin residents, those who answered that they do not wear shoes regularly had an elevated risk of infection (odds ratio = 2.50, 95% confidence interval = 1.03-6.08). CONCLUSIONS: This study showed that there is still an on-going transmission of strongyloidiasis in Lower Myanmar. It is highly desirable that the soil should be free of fecal contamination by improving the management of fecal waste. Meanwhile, health education to promote shoe-wearing would be beneficial to reduce the risk of transmission, especially for those who have frequent and intense contact with soil.

19.
Mol Biol Cell ; 28(3): 411-428, 2017 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-27932494

RESUMO

The ∼6000 species in phylum Apicomplexa are single-celled obligate intracellular parasites. Their defining characteristic is the apical complex-membranous and cytoskeletal elements at the apical end of the cell that participate in host-cell invasion. The apical complex of Toxoplasma gondii and some other apicomplexans includes a cone-shaped assembly, the conoid, which in T. gondii comprises 14 spirally arranged fibers that are nontubular polymers of tubulin. The tubulin dimers of the conoid fibers make canonical microtubules elsewhere in the same cell, suggesting that nontubulin protein dictates their special arrangement in the conoid fibers. One candidate for this role is TgDCX, which has a doublecortin (DCX) domain and a TPPP/P25-α domain, both of which are known modulators of tubulin polymer structure. Loss of TgDCX radically disrupts the structure of the conoid, severely impairs host-cell invasion, and slows growth. Both the conoid structural defects and the impaired invasion of TgDCX-null parasites are corrected by reintroduction of a TgDCX coding sequence. The nontubular polymeric form of tubulin found in the conoid is not found in the host cell, suggesting that TgDCX may be an attractive target for new parasite-specific chemotherapeutic agents.


Assuntos
Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Toxoplasma/metabolismo , Motivos de Aminoácidos , Animais , Citoesqueleto/metabolismo , Proteínas do Domínio Duplacortina , Microtúbulos/metabolismo , Organelas/metabolismo , Toxoplasma/genética , Toxoplasmose/genética , Toxoplasmose/metabolismo , Tubulina (Proteína)/metabolismo
20.
Trop Med Health ; 45: 20, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28814926

RESUMO

BACKGROUND: A prevalence study of Wuchereria bancrofti infection was carried out in 2014 at 4 study sites in northern Uganda using antigen and microfilaria tests. Each study site consists of a primary school and surrounding communities. These sites are inside the filariasis endemic area and have been covered by mass drug administration under the national elimination programme. However, no prevalence study had been conducted there before the present study. Without information on past and present endemicity levels, our study was meant to be an independent third-party investigation to know the latest filariasis situation. RESULTS: A total of 982 people including 570 schoolchildren (7-19 years) and 412 community people (7-25 years) were examined, all of them for filarial antigen and 695 for microfilariae. The study revealed that all subjects were negative by both methods. CONCLUSIONS: It was considered that annual mass drug administrations together with anti-malarial activities such as indoor residual spraying had contributed to the reduction of the filarial infection. However, based on the past data obtained near our study sites, we cannot exclude the possibility that filarial prevalence rates in our study sites were very low or even zero originally. During the study, we encountered several patients with lower leg edema and pachydermic (elephant skin-like), mossy skin lesion of the foot. Judging from clinical features and bare-footed life-style of people in the area, non-filarial elephantiasis, possibly podoconiosis, was suspected. This elephantiasis has been reported in areas where filariasis is not endemic.

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