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Sample records for inflamatoria por acanthamoeba

  1. Ceratite bilateral por Acanthamoeba: relato de caso Bilateral Acanthamoeba keratitis: case report

    Directory of Open Access Journals (Sweden)

    Wilson Nahmatallah Obeid

    2003-12-01

    Full Text Available A ceratite por Acanthamoeba é uma infecção ocular grave que, apesar dos recentes progressos no diagnóstico e tratamento, ainda provoca prolongada morbidade e perda da acuidade visual. Relatamos um caso de ceratite bilateral por Acanthamoeba em usuário de lentes de contato, que é o primeiro caso descrito na literatura brasileira.Acanthamoeba keratitis is a severe ocular infection which even with recent progress in diagnosis and treatment still causes long morbidity and loss of visual acuity. We describe a case of bilateral Acanthamoeba keratitis in a user of contact lenses, which is the first case discussed in Brazilian literature.

  2. Ceratite bilateral por Acanthamoeba: relato de caso Bilateral Acanthamoeba keratitis: case report

    OpenAIRE

    Wilson Nahmatallah Obeid; Rogério de Araújo; Luiz Antonio Vieira; Marco Antonio Campos Machado

    2003-01-01

    A ceratite por Acanthamoeba é uma infecção ocular grave que, apesar dos recentes progressos no diagnóstico e tratamento, ainda provoca prolongada morbidade e perda da acuidade visual. Relatamos um caso de ceratite bilateral por Acanthamoeba em usuário de lentes de contato, que é o primeiro caso descrito na literatura brasileira.Acanthamoeba keratitis is a severe ocular infection which even with recent progress in diagnosis and treatment still causes long morbidity and loss of visual acuity. W...

  3. Conjuntivite presumível por Acanthamoeba Conjunctivitis presumably due to Acanthamoeba

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    Ana Cristina de Carvalho Ruthes

    2004-12-01

    Full Text Available OBJETIVO: Abordar quatro casos de conjuntivite presumível por Acanthamoeba, descrevendo o diagnóstico, considerando sinais e sintomas e o tratamento instituído. MÉTODOS: Foram estudados casos de conjuntivite presumível por Acanthamoeba diagnosticados no Hospital de Olhos do Paraná (HOP, no período de setembro/1998 a janeiro/2002. Todos os olhos estudados foram submetidos a um protocolo de investigação que incluía exame oftalmológico completo, microbiologia e cultura de secreções conjuntivais. RESULTADOS: Os exames laboratoriais de microscopia e cultura do material colhido estes pacientes revelaram o diagnóstico de Acanthamoeba. A maioria dos pacientes referia olhos vermelhos e irritação ocular de longa data. Os autores encontraram correlação entre a cultura e o exame direto, em que se evidenciou a presença de cistos e trofozoítas do protozoário. CONCLUSÃO: Este é o primeiro relato de conjuntivite provavelmente por Acanthamoeba de acordo com a literatura revisada. Pacientes selecionados e refratários ao tratamento habitual de infecção ocular externa devem ser considerados para estudo laboratorial adequado à procura etiológica da doença.PURPOSE: To describe four cases of conjunctivitis presumably due to Acanthamoeba considering diagnosis, signs, symptoms and treatment. METHODS: We reviewed the medical records of all patients who presented a clinical diagnosis of Acanthamoeba conjunctivitis between September/1998 to January/2001 at the "Hospital de Olhos do Paraná (HOP". All eyes were submitted to a protocol of investigation that included ophthalmologic examination, microscopic examination and culture exams of conjunctival smears for adequate treatment. RESULTS: The laboratorial results of conjunctival smears revealed contamination with Acanthamoeba by direct examination and thereafter, confirmed by culture. The authors observed cysts and trophozoites of Acanthamoeba. CONCLUSION: This is the first report of

  4. Sindrome de respuesta inflamatoria fetal : adaptacion cardiovascular

    National Research Council Canada - National Science Library

    Arreaza-Graterol, Mortimer; Rojas-Barrera, Juan Diego; Molina-Giraldo, Saulo

    2011-01-01

    Introduccion: el sindrome de respuesta inflamatoria fetal es una condicion caracterizada por reaccion inflamatoria sistemica acompanada de alteraciones bioquimicas como la elevacion de la interleucina 6 (IL-6...

  5. Actividad inflamatoria en múltiples placas ateroscleróticas en pacientes fallecidos por infarto agudo de miocardio

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    Alejandro García Escudero

    2009-01-01

    Full Text Available IntroducciónEstudios clínicos y anatomopatológicos sugieren que los procesos inflamatorios tienen un papel importante en la inestabilidad de la placa aterosclerótica, dado que en pacientes con síndromes coronarios agudos se observan infiltrados inflamatorios difusos en las arterias coronarias.ObjetivosEvaluar y localizar la distribución de placas vulnerables e infiltrados inflamatorios en pacientes fallecidos por infarto agudo de miocardio.Material y métodosMediante microscopia óptica se estudiaron las arterias coronarias de 58 pacientes fallecidos por infarto de miocardio. En las arterias coronarias relacionadas con el infarto y en las no relacionadas se registraron las siguientes variables: presencia de trombo, rotura de placa, hemorragia intraplaca y presencia de infiltrado inflamatorio.ResultadosAl analizar las diferencias existentes entre las arterias responsables del infarto y en las no responsables se encontraron diferencias significativas con respecto a la presencia de trombo (69% versus 38%; p < 0,008 y de hemorragia intraplaca (69% versus 50%; p < 0,03. No se encontró una diferencia significativa entre la arteria responsable y la no responsable al evaluar la presencia de infiltrado inflamatorio en las placas ateroscleróticas (77% versus 71%; p = ns.ConclusiónEn el infarto agudo de miocardio se comprobó la presencia de actividad inflamatoria que afectaba a más de un vaso, con compromiso de otras arterias además de la responsable del infarto. Se detectó también accidente agudo de placa en más de una arteria coronaria.REV ARGENT CARDIOL 2009;77:81-87.

  6. Citologia de impressão no diagnóstico de infecção corneana por Acanthamoeba: relato de caso Diagnosis of Acanthamoeba corneal infection by impression cytology: case report

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    Jeison de Nadai Barros

    2007-03-01

    Full Text Available Relatamos três casos de infecção corneana por Acanthamoeba sp em que foi possível detectar cistos do microorganismo com a técnica de citologia de impressão. Três pacientes encaminhados ao Laboratório de Doenças Externas Oculares em 2004 com alterações superficiais da córnea foram submetidos ao exame de citologia de impressão para investigação da presença de cistos de Acanthamoeba sp. Duas amostras foram obtidas da córnea de cada paciente e coradas com PAS, hematoxilina e Papanicolaou. Investigação microbiológica de rotina e cultura também foram realizadas após raspado da córnea. O cultivo das amostras e a citologia de impressão foram positivas para Acanthamoeba sp em todos os pacientes, ao passo que os raspados corados com Giemsa foram positivos em dois casos. A citologia de impressão revelou cistos de Acanthamoeba sp entre feixe de células epiteliais corneanas e como células isoladas. Foram observados cistos no epitélio de um dos pacientes com a citologia de impressão após três meses de tratamento, enquanto o raspado foi negativo. No exame anatomopatológico observaram-se cistos no epitélio e estroma de uma córnea receptora de um dos pacientes após transplante. Neste estudo, a citologia de impressão detectou com sucesso cistos de Acanthamoeba sp em pacientes com acometimento epitelial. Por tratar-se de método não invasivo, a técnica pode ser usada para facilitar o diagnóstico mais precoce da infecção por Acanthamoeba, sendo útil também no acompanhamento do tratamento da doença.To describe three cases of corneal infection due to Acanthamoeba sp in which was possible to detect Acanthamoeba sp cysts by the corneal impression cytology technique. Three patients referred to the External Eye Disease Laboratory in 2004 with superficial corneal alterations were submitted to corneal specimen collection by impression cytology filter paper to investigate the presence of Acanthamoeba sp cysts. Two impression

  7. Estudo comparativo da resposta inflamatoria, em ratos, induzida por venenos de serpentes do genero Bothrops em estagios distintos de desenvolvimento

    OpenAIRE

    Susana Elisa Moreno

    1994-01-01

    Resumo: No presente trabalho se estuda a resposta inflamatória induzida por venenos de 3 serpentes de gênero Bothrops (B. jararaca, B. neuwiedi e B. moojeni) em estágios distintos de desenvolvimento, utilizando-se os modelos de peritonite e edema de pata. Os venenos de Bothrops moojeni (VBM), Bothrops jararaca (VBJ), Bothrops neuwiedi (VBN) induzem migração de neutrófilos, de maneira dose-dependente, para a cavidade peritoneal de ratos, com diferentes graus de intensidade, devido, provavelmen...

  8. Estudo comparativo da resposta inflamatoria, em ratos, induzida por venenos de serpentes do genero Bothrops em estagios distintos de desenvolvimento

    OpenAIRE

    Susana Elisa Moreno

    1994-01-01

    No presente trabalho se estuda a resposta inflamatória induzida por venenos de 3 serpentes de gênero Bothrops (B. jararaca, B. neuwiedi e B. moojeni) em estágios distintos de desenvolvimento, utilizando-se os modelos de peritonite e edema de pata. Os venenos de Bothrops moojeni (VBM), Bothrops jararaca (VBJ), Bothrops neuwiedi (VBN) induzem migração de neutrófilos, de maneira dose-dependente, para a cavidade peritoneal de ratos, com diferentes graus de intensidade, devido, provavelmente, a va...

  9. Acanthamoeba Keratitis FAQs

    Science.gov (United States)

    ... Amebic Encephalitis (GAE); Keratitis Parasites Home Share Compartir Acanthamoeba Keratitis FAQs On this Page What is Acanthamoeba ... developing Acanthamoeba keratitis? Infection of the Eye with Acanthamoeba What is Acanthamoeba keratitis? Acanthamoeba keratitis is a ...

  10. Acanthamoeba keratitis.

    Directory of Open Access Journals (Sweden)

    Nicholson A

    1995-07-01

    Full Text Available Acanthamoeba keratitis, common in soft lens wearers, is not commonly isolated. The reports of Acanthamoeba keratitis in Indian literature are few. We report here a case of Acanthamoeba Keratitis in a medical student using soft contact lenses, initially diagnosed and treated as a bacterial and later as a viral corneal ulcer, who responded extremely well to medical line of therapy.

  11. Análisis de la respuesta inflamatoria mediada por receptores tipo toll tras el consumo intensivo de alcohol o binge drinking

    OpenAIRE

    Costa Alba, Pilar

    2016-01-01

    [ES] En este estudio se plantea como hipótesis que aquellos pacientes con intoxicación etílica aguda tras un CIA o binge drinking presentan una activación de la respuesta inmune innata en sangre periférica frente a los sujetos sanos. Como consecuencia, estos pacientes presentarán un incremento de los niveles plasmáticos del ARNm de TLR y citocinas pro-inflamatorias. El objetivo del estudio pretende determinar la activación de la respuesta inmune innata en sangre periférica de pacientes con CI...

  12. Queratitis por Acanthamoeba sp.: primer caso confirmado por aislamiento y tipificación molecular en Bahía Blanca, provincia de Buenos Aires, Argentina Acanthamoeba sp. keratitis: first case confirmed by isolation and molecular typification in Bahía Blanca, Buenos Aires Province, Argentina

    Directory of Open Access Journals (Sweden)

    M. L. Gertiser

    2010-06-01

    Full Text Available Algunas especies del género Acanthamoeba provocan queratitis amebiana (QA, una infección de la córnea muy dolorosa, generalmente unilateral, con compromiso importante de la visión. Se presenta el caso de una mujer de 31 años usuaria de lentes de contacto con hábitos inadecuados de higiene y conservación de las lentes. La paciente consultó por enrojecimiento y dolor intenso en el ojo derecho, de dos meses de evolución. Luego del examen oftalmológico y ante la sospecha de una etiología parasitaria se realizó una biopsia de la zona lesionada, que se destinó al análisis bacteriológico y parasitológico en el laboratorio. También se remitieron los líquidos de lavado y el estuche de las lentes. No se detectó crecimiento bacteriano en las muestras. Los cultivos para la búsqueda de Acanthamoeba resultaron positivos en todos los especímenes analizados. Las amebas aisladas fueron clasificadas morfológica y molecularmente como pertenecientes al género Acanthamoeba. El presente trabajo muestra el primer caso de queratitis por Acanthamoeba de la ciudad de Bahía Blanca identificado por el laboratorio mediante procedimientos de alto valor diagnóstico.Some species of the Acanthamoeba genus cause keratitis, a very painful, most likely unilateral corneal infection , associated with eye and vision impairment. We here present a case of a 31-year-old female patient, a regular user of soft contact lenses without good practices of lens hygiene and handling. The patient attended medical consultation after two months of inflammation and pain in her right eye. After ophthalmological studies, and due to suspicion of a parasitic infection, a biopsy was performed and the sample submitted for bacteriological and parasitological analyses. Moreover, contact lens holders and lens cleaning solutions were studied. The samples yielded negative results for bacterial infection. However, cultivation of all samples showed the presence of amoeboid parasites

  13. Hiperplasia Fibrosa Inflamatoria: reporte de un caso

    OpenAIRE

    Casian Romero,A; Trejo Quiroz,P; De León Torres,C; D Carmona Ruiz

    2011-01-01

    Introducción: La Hiperplasia Fibrosa Inflamatoria es una alteración caracterizada por un agrandamiento del tejido conectivo; se considera una lesión reactiva ya que se presenta como respuesta a una irritante crónico local; es una lesión común en pacientes adolescentes y de edad adulta debido a que su etiología principal se asocia a maloclusión, al uso de aparatología protésica u ortodóntica mal ajustada, así como a la presencia de biofilm. Sin embargo, la población infantil, principalmente en...

  14. Acanthamoeba encephalitis

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    Kaushal V

    2008-01-01

    Full Text Available Central nervous system infection with free-living amoebae is rare. We present a fatal case of Acanthamoeba encephalitis in a 63-year-old female from India where acanthamoebae were demonstrated and cultured from CSF. In spite of treatment with amphotericin B, fluconazole and rifampicin the patient did not survive. Amoebic infection should be suspected in a patient of encephalitis of unexplained aetiology as timely diagnosis can lead to a favourable outcome.

  15. Acanthamoeba meningoencephalitis

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    S R Chandra

    2014-01-01

    Full Text Available Report of a case of young immunocompetent male adult with autopsy proven acanthamoeba meningoencephalitis. The patient presented with a protracted febrile illness of 3 months duration with features of meningoencephalitis, this was followed by rapid deterioration while on anti tuberculous therapy and steroids and ended fatally. His magnetic resonance imaging showed features of hemorrhagic meningoencephalitis and magnetic resonance spectroscopy showed choline peak. Autopsy revealed necrotizing meningoencephalitis and intraocular colonization due to acanthamoeba.

  16. [Acanthamoeba keratitis].

    Science.gov (United States)

    Bouheraoua, N; Labbé, A; Chaumeil, C; Liang, Q; Laroche, L; Borderie, V

    2014-10-01

    Early diagnosis and appropriate therapy are key elements for a good prognosis in Acanthamoeba keratitis (AK). AK should be considered in any case of corneal trauma complicated by exposure to soil or contaminated water, and in all contact lens (CL) wearers. A presumptive diagnosis of AK can be made clinically and with in vivo confocal microscopy, although a definitive diagnosis requires identification of Acanthamoeba on direct scraping, histology, or identification of Acanthamoeba DNA by polymerase chain reaction (PCR). We use cysticidal drugs for treating AK because encysted forms are more resistant than trophozoites to treatment. The treatment protocol used a biguanide (PHMB 0.02% or chlorhexidine 0.02%) and a diamidine (propamidine 0.1% or hexamidine 0.1%). New diagnostic modalities and more specific topical anti-amoebic treatments would substantially benefit patients with AK.

  17. Anemia y enfermedad inflamatoria intestinal Anemia and inflammatory bowel disease

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    F. de la Morena

    2008-05-01

    Full Text Available La anemia es una de las complicaciones más comunes de la enfermedad inflamatoria intestinal. La alta frecuencia de valores bajos de hemoglobina en estos enfermos provoca en muchas ocasiones una infravaloración por parte del médico de esta circunstancia, lo que se traduce en la falta de un tratamiento eficaz. Por otro lado, el carácter complejo de los mecanismos de producción de la anemia en la enfermedad inflamatoria intestinal con frecuencia plantea dudas acerca del tratamiento más adecuado. La identificación correcta de los pacientes con anemia así como la instauración del tratamiento más idóneo serán los dos pilares fundamentales para la mejoría de la calidad de vida de los enfermos. El uso correcto de los suplementos de hierro y las nuevas formulaciones de hierro parenteral, con o sin eritropoyetina asociada, han revolucionado nuestro abordaje de esta complicación evolutiva de la enfermedad inflamatoria intestinalAnemia is a most common complication of inflammatory bowel disease. A high frequency of low hemoglobin values in these patients often leads physicians to subestimate this condition, which translates into ineffective treatment. On the other hand, the complex nature of anemia-inducing mechanisms in inflammatory bowel disease frequently raises doubt about the most appropriate therapy. A correct identification of patients with anemia, and adequate therapy are the essential pillars for improved quality of life. The right use of iron supplementation, and novel parenteral iron formulations, either with or without associated erythropoietin, have revolutionized our approach of this complication in the course of inflammatory bowel disease

  18. Persistence of acanthamoeba antigen following acanthamoeba keratitis

    OpenAIRE

    Yang, Y; Matheson, M; Dart, J; Cree, I.

    2001-01-01

    AIM—To investigate the hypothesis that persistent corneal and scleral inflammation following acanthamoeba keratitis is not always caused by active amoebic infection but can be due to persisting acanthamoebic antigens
METHODS—24 lamellar corneal biopsy and penetrating keratoplasty specimens were obtained from 14 consecutive patients at various stages of their disease and divided for microscopy and culture. Histological sections were immunostained and screened for the presence of Acanthamoeba c...

  19. Osteoporosis y enfermedad inflamatoria intestinal Osteoporosis and inflammatory bowel disease

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    L. Menchén

    2005-02-01

    Full Text Available La enfermedad inflamatoria intestinal es una entidad crónica de etiología desconocida en cuyo desarrollo influyen múltiples variables, como son la susceptibilidad individual, genética e inmunológica, así como diferentes factores ambientales. Sus manifestaciones clínicas son muy variadas y pueden afectar a otros órganos diferentes del tracto digestivo, convirtiéndose por tanto en una enfermedad multisistémica. En los últimos años existe un interés creciente por una de estas manifestaciones, la osteoporosis y la osteopenia, que puede afectar hasta al 42% de los pacientes y condiciona un importante aumento de la morbilidad. La inactividad, el tratamiento corticoideo prolongado, las deficiencias nutricionales y la propia enfermedad pueden favorecer el desarrollo de esta complicación. En esta revisión se repasan aspectos clínicos y etiológicos de la osteoporosis asociada a la enfermedad inflamatoria intestinal y se ofrecen pautas para su diagnóstico y tratamiento.Inflammatory bowel disease is a chronic disease with an unknown ethiology although multiple factors intervene such as individual, genetic and immunologic susceptibility, as well as different environmental factors. Like other multisystemic diseases, its clinical manifestations are diverse and it may affect other organs besides the gastrointestinal tract. In the last few years there is a growing interest for one of these extraintestinal manifestations, osteoporosis and osteopenia that may affect up to 42% of patients and can condition an important increase in morbility. Inactivity, prolonged corticosteroid treatment, nutritional deficiencies and the disease per se have an important role in the development of this complication. This article reviews clinical and ethiological aspects of inflammatory bowel disease associated osteoporosis and offers a strategy for diagnosis and treatment.

  20. Biology and pathogenesis of Acanthamoeba

    OpenAIRE

    Siddiqui Ruqaiyyah; Khan Naveed

    2012-01-01

    Abstract Acanthamoeba is a free-living protist pathogen, capable of causing a blinding keratitis and fatal granulomatous encephalitis. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread and host susceptibility, and are highlighted together with potential therapeutic and preventative measures. The use of Acanthamoeba in the study of cellular differentiation mechanisms, motility and phagocytosis, bacterial pathogenesis and ev...

  1. Polirradiculopatía inflamatoria crónica sensitiva

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    Paz Zuberbuhler

    2013-06-01

    Full Text Available La polirradiculopatía inflamatoria crónica sensitiva es una entidad definida, frecuentemente subdiagnosticada y potencialmente tratable. Debe ser sospechada en pacientes con ataxia sensitiva, estudios de conducción nerviosa normales y una resonancia magnética que muestre engrosamiento y realce con gadolinio de las raíces lumbosacras. Presentamos el caso de un hombre de 57 años de edad con marcada ataxia sensitiva en pierna izquierda. Al examen físico presentaba fuerza conservada, reflejos osteotendinosos disminuidos, tacto fino y superficial reducidos por debajo de las rodillas; abatiestesia y apalestesia en ambos pies. Los estudios de conducción nerviosa eran normales, los potenciales evocados somatosensitivos tibiales con ausencia de respuesta bilateral. El líquido cefalorraquídeo presentaba hiperproteinorraquia sin células. La resonancia magnética mostró engrosamiento y realce con gadolinio de las raíces lumbosacras. El paciente fue tratado con inmunoglobulina endovenosa (IgEV a 2 g/kg durante 5 días, con buena respuesta. La evolución clínica, la hiperproteinorraquia, el realce de raíces en la resonancia magnética, la buena respuesta a la inmunoterapia y la exclusión de otras causas de ataxia sensitiva fueron compatibles con el diagnóstico de polirradiculopatía inflamatoria crónica sensitiva. Para el diagnóstico de esta enfermedad se requiere la identificación del compromiso aislado de las raíces sensitivas.

  2. Acanthamoeba keratitis in Pondicherry.

    Science.gov (United States)

    Parija, S C; Prakash, M R; Rao, V A; Vellaniparambil, R J

    2001-06-01

    Acanthamoeba keratitis is a potentially devastating infection of the cornea caused by the free-living amoebae, Acanthamoeba species. During the period from 1997 to 2000, a total of 136 corneal scrapings from clinically suspected cases were screened and examined for the presence of the Acanthamoeba. On examination of the direct smear by microscopy, 11 out of 136 cases were positive for Acanthamoeba. Eight patients were males and 3 were females. The age of these patients ranged from 15 to 57 years. All of these cases were agricultural workers who did not use contact lens. Four cases gave a history of injury to the eye and 1 patient gave a history of applying cow dung on the eye after the injury. Rest of the patients did not give any history of trauma or wearing contact lenses. The patients were treated with topical application of neosporin ointment. Many of our cases had complications such as poor vision (all 11 cases had 6/60 or less), scar formation (3 cases), opacity (5 cases) and corneal perforation (2 cases). This report documents for the first time the cases of Acanthamoeba keratitis in Pondicherry.

  3. Immunity to Acanthamoeba.

    Science.gov (United States)

    Ferrante, A

    1991-01-01

    Human serum contains antibodies, mainly of the IgM and IgG isotypes, to pathogenic species of Acanthamoeba. This, as well as the capacity of these amebas to activate complement via the alternative pathway, may be a first-line defense against acanthamoeba infections in humans. Both antibody and complement appear to be important in promoting recognition of these amebas by phagocytic cells such as neutrophils. However, killing of amebas by neutrophils is dependent on lymphokine/monokine priming of the neutrophil. This priming augments the respiratory-burst activity and release of lysosomal enzymes of neutrophils in their response to the ameba. The products of the oxygen-dependent respiratory burst appear to be of prime importance in the killing of this free-living ameba. Antibodies also may prevent tissue invasion by Acanthamoeba by inhibiting its adherence, phagocytic activity, and migration and by neutralizing cytopathogenic amebic agents. Studies on experimental Acanthamoeba infections in mice showed marked species and strain specificity with regard to induction of protection with amebic antigens. Immune compromise or, alternatively, invasion at unique body sites in healthy individuals may form the basis for human infection with Acanthamoeba.

  4. Enzimas proteolíticas relacionadas con la enfermedad periodontal inflamatoria

    OpenAIRE

    Bárbara E García Triana; Agustín Vicedo Tomey,; José C García Piñeiro; Alberto Saldaña Bernabeu

    1998-01-01

    La enfermedad periodontal inflamatoria ocasiona la destrucción de los tejidos que protegen y soportan al diente; es por eso de gran importancia el papel que pueden desempeñar las enzimas que sean capaces de degradar la matriz del tejido conectivo, como las enzimas proteolíticas. Existen evidencias de que las metaloproteinasas de la matriz, las proteasas leucocitarias y las bacterianas, pueden participar en la etiopatogenia de esta enfermedad. Su acción es regulada en los tejidos, por la prese...

  5. Variables psicológicas y calidad de vida en la enfermedad inflamatoria intestinal

    OpenAIRE

    Iglesias Rey, Marta

    2013-01-01

    Antecedentes: Bajo la denominación de enfermedad inflamatoria intestinal se incluyen un grupo de enfermedades crónicas del tracto gastrointestinal entre las que destacan por su gravedad y prevalencia, la enfermedad de Crohn y la colitis ulcerosa. El curso clínico se caracteriza por la aparición de brotes de reagudización de diversa gravedad y periodos de remisión en los que el paciente se encuentra clínicamente estable. El comienzo en edades tempranas, la cronicidad, necesidad de cirugía y ho...

  6. La reacción inflamatoria en la fisiopatogenia de la obesidad

    OpenAIRE

    Ma Victoria Domínguez García; Gerardo Gabriel Huitrón Bravo; Anaí A. Mendoza López

    2012-01-01

    La obesidad se caracteriza por inflamación crónica. Entre los biomarcadores de inflamación encontramos las citocinas producidas por los adipocitos y los macrófagos: TNF - a, IL - 6, leptina, adiponectina y resistina. La relación adipocito-macrófago impacta la respuesta inflamatoria observada en la obesidad, la cual tiene una relación directa con la cantidad de macrófagos acumulados en el tejido adiposo blanco, que contribuye a la producción de mediadores de la inflamación. En este documento s...

  7. Determinación de citoquinas en espondiloartropatías inflamatorias: valoración de la actividad clínico-biológica en la artropatía psoriásica

    OpenAIRE

    Turrión Nieves, Ana Isabel

    2015-01-01

    Las espondiloartritis o espondiloartropatías inflamatorias constituyen un grupo de enfermedades reumáticas inflamatorias que afectan fundamentalmente a la columna vertebral. De estas la espondilitis anquilosante (EAnq) y la artropatía psoriásica (Aps), son las afectaciones más frecuentes. Los niveles de citoquinas y/o reactantes de fase aguda son marcadores biológicos de actividad clínica inflamatoria. Por ello siendo estas moléculas cuantificables en el laboratorio, su determinación puntual ...

  8. Biology and pathogenesis of Acanthamoeba

    Directory of Open Access Journals (Sweden)

    Siddiqui Ruqaiyyah

    2012-01-01

    Full Text Available Abstract Acanthamoeba is a free-living protist pathogen, capable of causing a blinding keratitis and fatal granulomatous encephalitis. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread and host susceptibility, and are highlighted together with potential therapeutic and preventative measures. The use of Acanthamoeba in the study of cellular differentiation mechanisms, motility and phagocytosis, bacterial pathogenesis and evolutionary processes makes it an attractive model organism. There is a significant emphasis on Acanthamoeba as a Trojan horse of other microbes including viral, bacterial, protists and yeast pathogens.

  9. Pelvic inflammatory disease Enfermedad inflamatoria pélvica

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    Regla Fang Mederos

    2007-02-01

    Full Text Available

    A bibliographical revision on the main topics referred to the acute pelvic inflammatory disease which includes definition, risk factors, diagnosis, and treatment with the objective to create a supporting educational aid to the teaching of students of 4th and 6th year of the medicine and nursing specialty in their rounds on the services of gynecology and obstetrics.

    Revisión bibliográfica sobre los principales temas referidos a la enfermedad inflamatoria pélvica aguda que incluye concepto, factores de riesgo, cuadro clínico, diagnóstico y tratamiento, con el objetivo de crear un material de apoyo a la docencia de estudiantes de cuarto y sexto año de la especialidad de medicina y de enfermería en su rotación por los Servicios de Ginecología y Obstetricia.

  10. Acanthamoeba castellanii STAT Protein

    OpenAIRE

    Anna Kicinska; Jacek Leluk; Wieslawa Jarmuszkiewicz

    2014-01-01

    STAT (signal transducers and activators of transcription) proteins are one of the important mediators of phosphotyrosine-regulated signaling in metazoan cells. We described the presence of STAT protein in a unicellular, free-living amoebae with a simple life cycle, Acanthamoeba castellanii. A. castellanii is the only, studied to date, Amoebozoan that does not belong to Mycetozoa but possesses STATs. A sequence of the A. castellanii STAT protein includes domains similar to those of the Dictyos...

  11. Miopatía inflamatoria con compromiso inicial de músculos respiratorios y artritis reumatoidea

    OpenAIRE

    Martín Hunter; Irene Telias; Victoria Collado; Judith Sarano; Clarisa Álvarez; Juan Pablo Suárez

    2014-01-01

    Las miopatías inflamatorias constituyen un grupo heterogéneo de enfermedades musculares adquiridas de presentación subaguda, crónica y a veces aguda. Las entidades clínicas más frecuentes son la dermatomiositis, la polimiositis, la miositis necrotizante autoinmune y la miositis por cuerpos de inclusión. Suelen presentarse con debilidad muscular con predominio proximal y simétrica, pero rara vez comprometen los músculos respiratorios. Presentamos el caso de una mujer de 39 años con miopatía in...

  12. Campylobacter-Acanthamoeba interactions.

    Science.gov (United States)

    Vieira, Ana; Seddon, Alan M; Karlyshev, Andrey V

    2015-05-01

    Campylobacter jejuni is a foodborne pathogen recognized as the major cause of human bacterial enteritis. Undercooked poultry products and contaminated water are considered as the most important sources of infection. Some studies suggest transmission and survival of this bacterial pathogen may be assisted by the free-living protozoa Acanthamoeba. The latter is known to play the role of a host for various pathogenic bacteria, protecting them from harsh environmental conditions. Importantly, there is a similarity between the mechanisms of bacterial survival within amoebae and macrophages, making the former a convenient tool for the investigation of the survival of pathogenic bacteria in the environment. However, the molecular mechanisms involved in the interaction between Campylobacter and Acanthamoeba are not well understood. Whilst some studies suggest the ability of C. jejuni to survive within the protozoa, the other reports support an extracellular mode of survival only. In this review, we focus on the studies investigating the interaction between Campylobacter and Acanthamoeba, address some reasons for the contradictory results, and discuss possible implications of these results for epidemiology. Additionally, as the molecular mechanisms involved remain unknown, we also suggest possible factors that may be involved in this process. Deciphering the molecular mechanisms of pathogen-protozoa interaction will assist in a better understanding of Campylobacter lifestyle and in the development of novel antibacterial drugs.

  13. Miopatía inflamatoria con compromiso inicial de músculos respiratorios y artritis reumatoidea

    Directory of Open Access Journals (Sweden)

    Martín Hunter

    2014-10-01

    Full Text Available Las miopatías inflamatorias constituyen un grupo heterogéneo de enfermedades musculares adquiridas de presentación subaguda, crónica y a veces aguda. Las entidades clínicas más frecuentes son la dermatomiositis, la polimiositis, la miositis necrotizante autoinmune y la miositis por cuerpos de inclusión. Suelen presentarse con debilidad muscular con predominio proximal y simétrica, pero rara vez comprometen los músculos respiratorios. Presentamos el caso de una mujer de 39 años con miopatía inflamatoria inespecífica que presentó insuficiencia respiratoria secundaria a hipoventilación alveolar por debilidad muscular y requirió asistencia respiratoria mecánica. Respondió favorablemente y de forma rápida tras el tratamiento instaurado con inmunosupresores (corticoides y metotrexato e inmunoglobulina humana endovenosa. Se utilizó ventilación no invasiva como alternativa a la intubación orotraqueal con adecuada tolerancia.

  14. Treatment of Acanthamoeba keratitis.

    Science.gov (United States)

    Seal, David

    2003-08-01

    The treatment of Acanthamoeba keratitis has now been possible since the first successful therapy developed in the mid 1980s with a combination of propamidine 0.1% (Brolene) and neomycin 1%. However, only half the patients responded to this regimen as the cysts were often resistant to neomycin and relatively insensitive to propamidine. This led to research for better therapy, culminating in the mid 1990s with research in Glasgow demonstrating much increased effectiveness with use of the biguanide chlorhexidine 0.02% and in London and Bristol for similar effectiveness with the polymeric polyhexamethylene biguanide (PHMB) 0.02%. Both biguanides were combined with propamidine for enhanced effectiveness but were also shown to be effective as monotherapy. While this therapy inactivates the trophozoites and cysts in Acanthamoeba keratitis in the majority of patients (approximately 90%), there have been notable failures particularly when presentation is late with deep stromal infection. Additional highly acanthamoebicidal drugs are needed that can penetrate the stroma for synergistic action. This role may be taken up by certain antineoplastic drugs, such as alkylphosphocholine-1 (Miltefosine), that also have antiprotozoal activity.

  15. Modalidades de imagen en la evaluación de enfermedad inflamatoria intestinal

    Directory of Open Access Journals (Sweden)

    A. Guarnizo

    2016-07-01

    Full Text Available La enfermedad inflamatoria intestinal es una patología crónica, con una incidencia cada vez mayor. Dentro de este grupo de afecciones, la enfermedad de Crohn y la colitis ulcerosa son las más frecuentes. La enterografía por tomografía computada (ETC y por resonancia magnética (ERM son las modalidades de elección para la evaluación y seguimiento de la entidad, permitiendo examinar la apariencia de la mucosa, la pared intestinal, las manifestaciones extraintestinales y las complicaciones asociadas. La elección del estudio debe hacerse de acuerdo con la condición clínica de cada paciente.

  16. La reacción inflamatoria en la fisiopatogenia de la obesidad

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    Ma. Victoria Domínguez García

    2012-01-01

    Full Text Available La obesidad se caracteriza por inflamación crónica. Entre los biomarcadores de inflamación encontramos las citocinas producidas por los adipocitos y los macrófagos: TNF - a, IL - 6, leptina, adiponectina y resistina. La relación adipocito-macrófago impacta la respuesta inflamatoria observada en la obesidad, la cual tiene una relación directa con la cantidad de macrófagos acumulados en el tejido adiposo blanco, que contribuye a la producción de mediadores de la inflamación. En este documento se hará una revisión de las principales moléculas involucradas en la generación de inflamación en la obesidad, y la relación entre macrófagos y adipocitos.

  17. Experiencia con rituximab en miopatía inflamatoria idiopática refractaria

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    Elmer R. García-Salazar

    2013-10-01

    Full Text Available Se describe las características clínicas y de laboratorio de dos pacientes que recibieron rituximab por miopatía inflamatoria idiopática (MII. Ellas eran refractarias a tratamiento convencional con DARMES, por lo que recibieron rituximab 1 gramo cada 14 días, en dos infusiones en ciclo semestral. En las historias clínicas se obtuvo los datos clínicos de fuerza muscular proximal, lesiones cutáneas patognomónicas, elevación de CPK, TGO, DHL y VSG, resultados de electromiografía, biopsia muscular y de piel. Ninguno de los dos casos presentó reacción medicamentosa ni infecciones durante y posterior a las infusiones. Rituximab mostró efectividad en la respuesta clínica y enzimática en estas pacientes con dermatomiositis refractarias a corticoides y DARMES tradicionales.

  18. Amiloidosis secundaria en la enfermedad inflamatoria intestinal Secondary amyloidosis in Chrohn's disease

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    S. Seijo Ríos

    2008-12-01

    Full Text Available La amiloidosis es una entidad clínica que se produce a consecuencia del depósito a nivel extracelular de un material proteico amorfo, causando una desorganización de la arquitectura normal de múltiples órganos y tejidos y, por tanto, una alteración funcional de los mismos. La amiloidosis secundaria es una complicación infrecuente pero muy grave que aparece en el contexto de neoplasias, enfermedades infecciosas e inflamatorias de curso crónico, como es el caso de la enfermedad inflamatoria intestinal, principalmente enfermedad de Crohn, ensombreciendo el pronóstico de estos pacientes. A continuación presentamos dos casos clínicos correspondientes a dos pacientes con enfermedad de Crohn que desarrollaron amiloidosis secundaria.Amyloidosis is a clinical entity that results from the deposition of an extracellular protein material that causes disruption in the normal architecture of multiple organs and tissues, and impairs their function. Secondary amyloidosis is a rare but serious complication that may worsen the prognosis of patients with cancer, infection or chronic inflammatory disease, including inflammatory bowel disease, particularly Crohn's disease. We report two cases of Crohn's disease associated with secondary amyloidosis.

  19. La Peroxidación lipídica en la enfermedad periodontal inflamatoria experimental

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    Bárbara E García Triana

    1998-04-01

    Full Text Available La peroxidación de los lípidos de las membranas de los tejidos periodontales, parece estar implicada en la etiopatogenia de la enfermedad periodontal inflamatoria. En este estudio utilizamos perros Beagles, a los cuales se les provocó la enfermedad, por ligadura de los dientes de una hemiarcada, mientras la hemiarcada opuesta se empleó como control. Al final del experimento, las variables estudiadas (profundidad del surco gingival, altura ósea y niveles de malonil-dialdehído fueron significativamente superiores en la hemiarcada ligada, con respecto a la no ligada. Concluimos que el uso de este modelo es factible en nuestras condiciones. Estos resultados apoyan la teoría de la participación de la peroxidación lipídica en la etiopatogenia de la enfermedad periodontal inflamatoria.The lipid peroxidation of the periodontal membranes seems to be involved in the pathogenesis of the periodontal disease. We used for this study Beagles dogs which were caused such a disease by applying a ligature to a dental hemiarch teeth whereas the opposite hemiarch was used as a control. At the end of the experiment, the studied variables (gingival sulcus depth, bone height and malonyl-dialdehyde levels were significantly higher in the ligated hemiarch. We concluded that the use of this model is feasible in our conditions. These results back up the theory of the role of lipid peroxidation in the pathogenesis of the periodontal diseases.

  20. Pleuresia inflamatoria en la infancia: Conducta quirúrgica

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    Arturo L. Delgado Delgado

    2003-04-01

    Full Text Available La neumonía complicada por pleuresía ha sido siempre un proceso que ha afectado la morbilidad y mortalidad en la edad pediátrica y en ocasiones necesita de algún proceder quirúrgico para su solución. La decisión inicial de si realizar solo una punción pleural o agregarle una pleurostomía mínima, puede ser controversial, por lo que se decidió realizar un estudio de todos aquellos niños que fueron ingresados en el Hospital Pediátrico Universitario de San Miguel del Padrón con neumonía complicada por pleuresía inflamatoria (pleuresía durante un proceso inflamatorio pulmonar, a los cuales hubo que realizarles algún proceder quirúrgico para la solución de su derrame, en el período de enero de 2000 a enero de 2001; se estudiaron los expedientes clínicos de 10 pacientes; se les conformaron tablas y figuras. El grupo de edad más afectado fue el de 1 a 5 años con un 40 %. El sexo no tuvo preponderancia y la localización más frecuente del proceso inflamatorio (90 % fueron el pulmón derecho, y entre estas la base derecha con un 40 %. Al 50 % de los pacientes se le realizó solamente una punción pleural y a la otra mitad se le añadió una pleurostomía, y tenía el 30 % en este último grupo más de 150 mL en la punción inicial; a 5 pacientes hubo que realizarles una pleurostomía mínima. Con estos resultados se consideró que en el momento de la punción pleural inicial el hallazgo macroscópico de aspecto fibrino purulento es indicativo de pleurostomía, así como este aspecto en el líquido de una segunda punción de ser necesario ésta y una celularidad mayor de 175 ´ 106, y una glucosa menor de 1 mmol/L en el microscópico del líquido de la primera, si se hace necesario realizar una segunda punción o los mismos resultados del líquido proveniente de esta segunda o cualquier otra punción.Pneumonia complicated due to pleurisy has always been a process affecting morbidity and mortality at the pediatric age and on

  1. Enzimas proteolíticas relacionadas con la enfermedad periodontal inflamatoria

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    Bárbara E. García Triana

    1998-08-01

    Full Text Available La enfermedad periodontal inflamatoria ocasiona la destrucción de los tejidos que protegen y soportan al diente; es por eso de gran importancia el papel que pueden desempeñar las enzimas que sean capaces de degradar la matriz del tejido conectivo, como las enzimas proteolíticas. Existen evidencias de que las metaloproteinasas de la matriz, las proteasas leucocitarias y las bacterianas, pueden participar en la etiopatogenia de esta enfermedad. Su acción es regulada en los tejidos, por la presencia de inhibidores específicos, de manera que un desbalance proteasas-inhibidores a favor de los primeros, conduciría a la destrucción de las proteínas de la matriz del tejido conectivo. A su vez, en la actividad proteolítica influyen diferentes factores, que de manera global, inducen un fenotipo degradativo o formativo, y que por lo tanto, podrían estar involucrados en la etiopatogenia de la enfermedad periodontal inflamatoria.The periodontal disease brings about the destruction of the protective and supporting tissues of the teeth, therefore, the role of enzymes capable of degrading connective tissue matrix is of great importance. There are evidences of the possible involvement of the matrix metalloproteinases, leukocyte proteases and bacterial proteases in the pathogenesis of such disease. Their action is controlled by specific inhibitors in the tissues. This means that any protease-inhibitor imbalance favouring the presence of proteases may lead to the destruction of matrix connective tissue. In turn, proteolytic activity is influenced by different factors that globally induce a degradative or formative phenotype, and thus, may be involved in the pathogenesis of the periodontal disease.

  2. Hipnosis clínica en un caso de enfermedad inflamatoria intestinal

    Directory of Open Access Journals (Sweden)

    J. C. FERNÁNDEZ MÉNDEZ

    2000-01-01

    Full Text Available Un sujeto de veinte años con una enfermedad inflamatoria intestinal fue tratado con hipnosis. Hasta donde sabemos es el primer informe que documenta el empleo de la hipnosis clínica en un paciente con enfermedad de Crohn, para reducir el estrés y afrontar las demandas originadas por la patología digestiva. Además, otro aspecto importante en el presente trabajo, es el registro exhaustivo y consecutivo de la sintomatología gastrointestinal, un punto débil en la gran mayoría de los trabajos precedentes. Al finalizar la intervención psicológica disminuyó notablemente el nivel de intensidad del estrés, el número de deposiciones diarias así como la intensidad del dolor abdominal. Estos resultados se mantuvieron a lo largo del seguimiento.

  3. Acanthamoeba castellanii STAT protein.

    Directory of Open Access Journals (Sweden)

    Anna Kicinska

    Full Text Available STAT (signal transducers and activators of transcription proteins are one of the important mediators of phosphotyrosine-regulated signaling in metazoan cells. We described the presence of STAT protein in a unicellular, free-living amoebae with a simple life cycle, Acanthamoeba castellanii. A. castellanii is the only, studied to date, Amoebozoan that does not belong to Mycetozoa but possesses STATs. A sequence of the A. castellanii STAT protein includes domains similar to those of the Dictyostelium STAT proteins: a coiled coil (characteristic for Dictyostelium STAT coiled coil, a STAT DNA-binding domain and a Src-homology domain. The search for protein sequences homologous to A. castellanii STAT revealed 17 additional sequences from lower eukaryotes. Interestingly, all of these sequences come from Amoebozoa organisms that belong to either Mycetozoa (slime molds or Centramoebida. We showed that there are four separated clades within the slime mold STAT proteins. The A. castellanii STAT protein branches next to a group of STATc proteins from Mycetozoa. We also demonstrate that Amoebozoa form a distinct monophyletic lineage within the STAT protein world that is well separated from the other groups.

  4. Acanthamoeba castellanii STAT protein.

    Science.gov (United States)

    Kicinska, Anna; Leluk, Jacek; Jarmuszkiewicz, Wieslawa

    2014-01-01

    STAT (signal transducers and activators of transcription) proteins are one of the important mediators of phosphotyrosine-regulated signaling in metazoan cells. We described the presence of STAT protein in a unicellular, free-living amoebae with a simple life cycle, Acanthamoeba castellanii. A. castellanii is the only, studied to date, Amoebozoan that does not belong to Mycetozoa but possesses STATs. A sequence of the A. castellanii STAT protein includes domains similar to those of the Dictyostelium STAT proteins: a coiled coil (characteristic for Dictyostelium STAT coiled coil), a STAT DNA-binding domain and a Src-homology domain. The search for protein sequences homologous to A. castellanii STAT revealed 17 additional sequences from lower eukaryotes. Interestingly, all of these sequences come from Amoebozoa organisms that belong to either Mycetozoa (slime molds) or Centramoebida. We showed that there are four separated clades within the slime mold STAT proteins. The A. castellanii STAT protein branches next to a group of STATc proteins from Mycetozoa. We also demonstrate that Amoebozoa form a distinct monophyletic lineage within the STAT protein world that is well separated from the other groups.

  5. Acanthamoeba everywhere: high diversity of Acanthamoeba in soils.

    Science.gov (United States)

    Geisen, Stefan; Fiore-Donno, Anna Maria; Walochnik, Julia; Bonkowski, Michael

    2014-09-01

    Acanthamoeba is a very abundant genus of soil protists with fundamental importance in nutrient cycling, but several strains can also act as human pathogens. The systematics of the genus is still unclear: currently 18 small-subunit (SSU or 18S) ribosomal RNA sequence types (T1-T18) are recognized, which sometimes contain several different morphotypes; on the other hand, some morphological identical strains belong to different sequence types, sometimes appearing in paraphyletic positions. In this study, we cultivated 65 Acanthamoeba clones from soil samples collected under grassland at three separate locations in the Netherlands, in Sardinia and at high altitude mountains in Tibet. We obtained 24 distinct partial sequences, which predominantly grouped within sequence type T4 followed by T2, T13, T16 and "OX-1" (in the T2/T6 clade). Our sequences were 98-99% similar, but none was identical to already known Acanthamoeba sequences. The community composition of Acanthamoeba strains differed between locations, T4 being the dominant sequence type in Sardinia and Tibet, but represented only half of the clones from soils in the Netherlands. The other half of clones from the Dutch soils was made up by T2, T16 and "OX-1", while T13 was only found in Sardinia and Tibet. None of the sequences was identical between localities. Several T4 clones from all three localities and all T13 clones grew at 37 °C while one T4 clone was highly cytopathogenic.

  6. Protease activities of Acanthamoeba polyphaga and Acanthamoeba castellanii.

    Science.gov (United States)

    Serrano-Luna, José de Jesús; Cervantes-Sandoval, Isaac; Calderón, Jesús; Navarro-García, Fernando; Tsutsumi, Victor; Shibayama, Mineko

    2006-01-01

    Acanthamoeba spp. are free-living amoebae that cause amoebic granulomatous encephalitis, skin lesions, and ocular amoebic keratitis in humans. Several authors have suggested that proteases could play a role in the pathogenesis of these diseases. In the present work, we performed a partial biochemical characterization of proteases in crude extracts of Acanthamoeba spp. and in conditioned medium using 7.5% SDS-PAGE copolymerized with 0.1% m/v gelatin as substrate. We distinguished a total of 17 bands with proteolytic activity distributed in two species of Acanthamoeba. The bands ranged from 30 to 188 kDa in A. castellanii and from 34 to 144 kDa in A. polyphaga. Additionally, we showed that the pattern of protease activity differed in the two species of Acanthamoeba when pH was altered. By using protease inhibitors, we found that the proteolytic activities belonged mostly to the serine protease family and secondly to cysteine proteases and that the proteolytic activities from A. castellanii were higher than those in A. polyphaga. Furthermore, aprotinin was found to inhibit crude extract protease activity on Madin-Darby canine kidney (MDCK) monolayers. These data suggest that protease patterns could be more complex than previously reported.

  7. Recomendaciones de vacunación en pacientes con enfermedad inflamatoria intestinal (EII

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    C. Rodríguez

    2013-04-01

    Full Text Available El concepto de enfermedad inflamatoria intestinal (EII engloba tres entidades: la colitis ulcerosa (CU, la enfermedad de Crohn (EC y la colitis inclasificable (CI. Estas enfermedades tienen en común el curso crónico y recidivante, alternando épocas de marcada actividad inflamatoria con otras quiescentes, en las que el paciente permanece asintomático. Durante muchos años la base del tratamiento, sobre todo en las fases agudas, se basó en el uso de corticoides. Sin embargo, a lo largo de las últimas décadas hemos asistido a avances importantes desde el punto de vista terapéutico. Así se estima que, a lo largo de la evolución de la enfermedad, el 80% de los pacientes van a precisar corticoides, el 40% inmunomoduladores (IMM y hasta un 20% necesitará un fármaco biológico para el control de su enfermedad. Si bien todo ello se acompaña de una mejora en la calidad de vida, disminuyendo la necesidad de ingresos e intervenciones quirúrgicas, su uso implica también un incremento en el riesgo de sufrir infecciones, bien por gérmenes habituales en la comunidad o por gérmenes oportunistas. Las infecciones, además, son causa de morbimortalidad asociada a EII y algunas de ellas son prevenibles con vacunas, de ahí la importancia que los programas de vacunación están adquiriendo en este grupo de pacientes. Presentamos una revisión de la literatura al respecto y proponemos unas recomendaciones de vacunación para los pacientes diagnosticados de EII.

  8. Manifestaciones reumatológicas de la enfermedad inflamatoria intestinal Rheumatologic manifestations of inflammatory bowel disease

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    Octavio Germán Muñoz Maya

    2006-01-01

    Full Text Available La enfermedad inflamatoria intestinal (EII se caracteriza por la activación inapropiada del sistema inmune de la mucosa intestinal y sus dos formas de presentación son: la colitis ulcerativa y la enfermedad de Crohn. Las manifestaciones extraintestinales se presentan hasta en el 36% de los pacientes y pueden comprometer cualquier órgano o sistema. La disfunción inmune se caracteriza por el desequilibrio entre los mediadores proinflamatorios y los antinflamatorios y se expresa como una enfermedad sistémica. Las manifestaciones reumatológicas asociadas a la EII son de tres tipos: la artritis periférica, la espondiloartropatía y una tercera categoría que incluye lesiones dérmicas, oftálmicas y del metabolismo óseo, entre otras. El manejo de estas manifestaciones se basa en la terapia sistémica para el control de la actividad inflamatoria local utilizando esteroides, derivados de la 5-ASA, inmunomoduladores y, en los últimos años, terapia anti-TNF. The main feature of inflammatory bowel disease (IBD is the continuous activation of the mucosa-associated immune system; the disease has two major forms of presentation: ulcerative colitis and Crohn´s disease. The extraintestinal manifestations are present in 36% of patients, and any organ can be affected. There is an imbalance between proinflammatory and antinflammatory cytokines leading to a systemic disease. The rheumatologic manifestations of the IBD are: Peripheral arthritis, spondyloarthropathy and a third category that includes dermic and ocular lesions as well as metabolic bone disease. Control of the extraintestinal manifestations is based on systemic therapy with steroids, 5-ASA derivatives and biological anti-TNF therapy.

  9. Impacto de la fatiga en la percepción de salud en la enfermedad inflamatoria intestinal.

    OpenAIRE

    Castillo Cejas, María Dolores

    2011-01-01

    La salud de los pacientes con enfermedad inflamatoria intestinal (EII) está afectada por la fatiga, que condiciona una baja calidad de vida. Este estudio administró cuestionarios de calidad de vida y fatiga a 110 pacientes con EII, cuantificando fatiga y correlacionándola con la actividad, se determinó la afectación de la calidad de vida. Los resultados mostraron relación inversa entre fatiga y actividad de EII, con diferencias estadísticamente significativas según brote o remisión de la enfe...

  10. Enfermedad inflamatoria intestinal : papel del análisis molecular de los virus de la familia herpes sobre material tisular

    OpenAIRE

    Azueta Etxebarria, Ainara

    2014-01-01

    RESUMEN: Los virus de la familia del herpes humano afectan al hombre de manera ubicua lo que hace discutible su papel patogenético en ciertas enfermedades a pesar de demostrarse su presencia. Por otro lado, la dificultad para interpretar las técnicas de laboratorio disponibles para su detección y llegar a un diagnóstico preciso impide en la mayor parte de los casos la detección precoz de la infección y el tratamiento eficaz. Existen casos de enfermedad inflamatoria intestinal en los que...

  11. Tratamiento psicológico protocolizado en grupo de las enfermedades inflamatorias intestinales: efectos sobre la calidad de vida

    OpenAIRE

    Miguel Ángel Díaz-Sibaja; María Isabel Comeche-Moreno; Blanca Mas-Hesse; Marta Díaz García; Miguel Ángel Vallejo Pareja

    2008-01-01

    La enfermedad inflamatoria intestinal (EII) es una enfermedad crónica que afecta notablemente a la calidad de vida del paciente. El principal objetivo de este estudio fue demostrar la eficacia de un programa de tratamiento psicológico protocolizado para mejorar la calidad de vida de estos pacientes. La muestra estuvo constituida por 57 pacientes diagnosticados de EII (33 programa de tratamiento en grupo y 24 grupo control en lista de espera). Las variables dependientes fueron: síntomas digest...

  12. Tratamiento, mediante bloqueo de moléculas CD20 con Rituximab, en miopatías Inflamatorias Idiopáticas refractarias a tratamiento convencional

    OpenAIRE

    Chinchilla Palomares, Eduardo

    2012-01-01

    Las miopatías inflamatorias idiopáticas (MII) constituyen un grupo heterogéneo de enfermedades adquiridas, de probable mecanismo inflamatorio autoinmune, que se caracterizan por debilidad muscular e infiltrado inflamatorio local o difuso, junto con necrosis de las fibras musculares, en la biopsia muscular. Afectan preferentemente a la musculatura estriada. Dentro de este grupo se incluyen tres variantes principales: la dermatomiositis (DM), la polimiositis (PM) y la miositis con cuerpos de in...

  13. Modelos animales para el estudio de la respuesta inflamatoria sistémica y de nutrición parenteral Animal models for the study of systemic inflammatory response and parenteral nutrition

    OpenAIRE

    J. M. Morán Penco

    2007-01-01

    El Síndrome de Respuesta Inflamatoria Sistémica (SRIS) parece desencadenarse por la activación de un tipo de receptores llamados Toll-Like, propios de las células de respuesta inflamatoria y que, a través de señales citosólicas específicas, producen una cascada de reacciones que activan las Citocinas, Factores de Crecimiento y otros mediadores inflamatorios. En este trabajo, repasamos y discutimos varias clasificaciones de Modelos Animales para el estudio del SRIS y proponemos que estos model...

  14. Photochemotherapeutic strategy against Acanthamoeba infections.

    Science.gov (United States)

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Anwar, Ayaz; Shah, Muhammad Raza; Khoja, Shahrukh; Khan, Naveed Ahmed

    2015-01-01

    Acanthamoeba is a protist pathogen that can cause serious human infections, including blinding keratitis and a granulomatous amoebic encephalitis that almost always results in death. The current treatment for these infections includes a mixture of drugs, and even then, a recurrence can occur. Photochemotherapy has shown promise in the treatment of Acanthamoeba infections; however, the selective targeting of pathogenic Acanthamoeba has remained a major concern. The mannose-binding protein is an important adhesin expressed on the surface membranes of pathogenic Acanthamoeba organisms. To specifically target Acanthamoeba, the overall aim of this study was to synthesize a photosensitizing compound (porphyrin) conjugated with mannose and test its efficacy in vitro. The synthesis of mannose-conjugated porphyrin was achieved by mixing benzaldehyde and pyrrole, yielding tetraphenylporphyrin. Tetraphenylporphyrin was then converted into mono-nitrophenylporphyrin by selectively nitrating the para position of the phenyl rings, as confirmed by nuclear magnetic resonance (NMR) spectroscopy. The mono-nitrophenylporphyrin was reduced to mono-aminophenylporphyrin in the presence of tin dichloride and confirmed by a peak at m/z 629. Finally, mono-aminoporphyrin was conjugated with mannose, resulting in the formation of an imine bond. Mannose-conjugated porphyrin was confirmed through spectroscopic analysis and showed that it absorbed light of wavelengths ranging from 425 to 475 nm. To determine the antiacanthamoebic effects of the derived product, amoebae were incubated with mannose-conjugated porphyrin for 1 h and washed 3 times to remove extracellular compound. Next, the amoebae were exposed to light of the appropriate wavelength for 1 h. The results revealed that mannose-conjugated porphyrin produced potent trophicidal effects and blocked excystation. In contrast, Acanthamoeba castellanii incubated with mannose alone and porphyrin alone did not exhibit an antiamoebic effect

  15. Pathogenicity, morphology, and differentiation of Acanthamoeba.

    Science.gov (United States)

    Khan, N A

    2001-12-01

    Acanthamoeba keratitis is sight threatening corneal infection caused by pathogenic Acanthamoeba. Previous studies have shown the genotypic differences between pathogenic and non-pathogenic species/strains of Acanthamoeba. In this study, we examined the morphological differences between pathogenic and non-pathogenic species/strains using scanning electron microscopy. Pathogenic Acanthamoeba exhibited higher number of acanthopodia (structures associated with the binding of amoeba to the target cells) as compared to non-pathogens. In addition, interactions of amoeba with the corneal epithelial cells were studied. Only pathogenic amoeba exhibited adhesion to epithelial cells. Further results indicated that phagocytosis occurs in the pathogenic amoeba by the formation of amoebastome (characteristic of amoeba phagocyte). This study showed that Acanthamoeba phagocytosis may be both an efficient means of obtaining nutrients for the amoeba and a significant factor in the pathogenesis of Acanthamoeba infections.

  16. Acanthamoeba sp.: un caso de queratitis no relacionada con el uso de lentes de contacto Acanthamoeba sp.: a case report in a non-contact lens wearer

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    Claudia Menghi

    2012-12-01

    Full Text Available La queratitis por Acanthamoeba sp. generalmente se relaciona con el uso de lentes de contacto y en menor proporción se vincula con el contacto con agua contaminada. Se caracteriza por una disminución de la capacidad visual y la presencia de un fuerte dolor ocular. al examen clínico, puede confundirse con una infección herpética. Si la infección no es diagnosticada y tratada a tiempo en forma adecuada, puede culminar en la perforación de la córnea y, eventualmente, en la pérdida del ojo. en este trabajo se informa uno de los pocos casos registrados de queratitis por Acanthamoeba sp. no relacionada con el uso de lentes de contacto.Acanthamoeba sp. keratitis is generally related to wearing contact lenses, and, to a lesser extent, to contaminated water. it is characterized by reduced visual capacity and the presence of severe ocular pain. Clinically, it can be mistaken for a herpes infection. if it is not diagnosed, and timely and adequately treated, it can result in corneal perforation and, eventually, in vision loss. One of the few registered cases of Acanthamoeba sp. keratitis not related to the use of contact lenses is herein reported.

  17. Bilateral Acanthamoeba ulcer in a user of disposable soft contact lenses: a tragic incident or a consequence of the aggressive policy of soft contact lens trading? Úlcera bilateral por Acanthamoeba em usuária de lentes de contato gelatinosas descartáveis: um incidente trágico ou uma conseqüência da política agressiva de venda de lentes de contato gelatinosas?

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    Sidney Júlio de Faria e Sousa

    2008-06-01

    Full Text Available This is the report of a case of bilateral Acanthamoeba keratitis in a 19-year-old woman who bought a pair of disposable soft contact lenses in a boutique. She wore this same pair of lenses for 3 months daily without the appropriate care. This led to bilateral corneal transplantation with cataract extraction and also trabeculectomy in the right eye. When last seen, both grafts were crystal clear but the visual acuities were far from satisfactory. She also had bilateral secondary glaucoma, barely controlled by topical medication. Actually, the physical features and the wearing time characteristics of the disposable soft contact lenses created unprecedented difficulties to the medical surveillance of their wearers. Without the right assistance they tend to become careless regarding routine cleaning. They also feel free to buy less expensive lenses, to use saline instead of lens solutions, to violate the limits of wearing time and to extend the use over the sleeping period. Additionally, the aggressive marketing and the wide distribution of these lenses increase the chances that economically or educationally unprepared people will acquire them. The question that remains is: Is the present case an accidental event or an example of what is likely to happen in the future if the indiscriminate selling of disposable soft contact lenses continues to evolve?Este é o relato de uma paciente do sexo feminino, de 19 anos, com ceratite bilateral por Acanthamoeba, devido ao uso de lentes de contato descartáveis, adquiridas em uma butique. Fez uso do mesmo par de lentes durante três meses, sem os cuidados adequados. Foi submetida a transplante penetrante de córnea e facectomia bilateral, além de trabeculectomia no olho direito. Quando vista pela última vez, os enxertos estavam transparentes, mas a visão de ambos os olhos não era satisfatória. Ela também apresentava glaucoma secundário bilateral, controlado precariamente com medicação tópica. Ocorre

  18. Manifestaciones orales y periodontales de la enfermedad inflamatoria intestinal

    OpenAIRE

    2001-01-01

    La Enfermedad Inflamatoria Intestinal (EII) incluye la enfermedad de Crohn y la colitis ulcerosa. Numerosos estudios relacionan dichas afectaciones con la manifestación de determinadas lesiones orales. Respecto a la enfermedad periodontal algunos autores la correlacionan con la EII tanto en su prevalencia como en su grado de actividad. Se ha apuntado la alteración de los PMN como posible factor común en la patogénesis de ambas entidades. El propósito de nuestro artículo es revisar las manifes...

  19. Hiperplasia fibrosa inflamatoria y posibles factores asociados en adultos mayores

    OpenAIRE

    Rodríguez Zegarra, Anita F.; Facultad de Estomatología, Universidad Peruana Cayetano Heredia. Lima,; Sacsaquispe-Contreras, Sonia; Facultad de Estomatología, Universidad Peruana Cayetano Heredia. Lima,

    2014-01-01

    El propósito del estudio fue determinar la prevalencia de hiperplasia fibrosa inflamatoria (HFI)asociada al uso de prótesis dental removible y sus factores asociados como: sexo, grado deinstrucción, edad, compromiso sistémico, estado nutricional e higiene oral del sujeto; tipo ytiempo de uso de la prótesis, tamaño y ubicación de la lesión en adultos mayores residentes delAlbergue Central “Ignacia Rodulfo Vda. de Canevaro”. Se evaluaron 91 sujetos mayores de 60años (80,67+7,65), que portaban p...

  20. Granulomatous inflammation in Acanthamoeba sclerokeratitis

    Directory of Open Access Journals (Sweden)

    Samrat Chatterjee

    2013-01-01

    Full Text Available This report describes the histopathological findings in a patient with Acanthamoeba sclerokeratitis (ASK. A 58-year-old patient with ASK underwent enucleation and sections of the cornea and sclera were subjected to histopathology and immunohistochemistry with monoclonal mouse antihuman antibodies against T cell CD3 and B cell CD20 antigens. Hematoxylin and Eosin stained sections of the cornea revealed epithelial ulceration, Bowman′s membrane destruction, stromal vascularization, infiltration with lymphocytes, plasma cells, and granulomatous inflammation with multinucleated giant cells (MNGC. The areas of scleritis showed complete disruption of sclera collagen, necrosis and infiltration with neutrophils, macrophages, lymphocytes, and granulomatous inflammation with MNGC. No cyst or trophozoites of Acanthamoeba were seen in the cornea or sclera. Immunophenotyping revealed that the population of lymphocytes was predominantly of T cells. Granulomatous inflammation in ASK is probably responsible for the continuance and progression of the scleritis and management protocols should include immunosuppressive agents alongside amoebicidal drugs.

  1. Histopathological examination of Acanthamoeba sclerokeratitis

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    Kato K

    2014-01-01

    Full Text Available Kumiko Kato,1 Koji Hirano,2 Tetsuro Nagasaka,3 Koichi Matsunaga,1 Yuko Takashima,1 Mineo Kondo1 1Department of Ophthalmology, Mie University School of Medicine, Tsu, Japan; 2Department of Ophthalmology, Fujita Health University, Nagoya, Japan; 3Department of Medical Laboratory Science, Nagoya University Graduate School of Medicine, Nagoya, Japan Purpose: To report the histopathological findings in a case of severe Acanthamoeba sclerokeratitis (ASK. Patient and methods: A 46-year-old patient was referred to the Department of Ophthalmology of Mie University Hospital because of a severe corneal ulcer of the right eye of 6 months' duration. Our initial examination showed a ring-shaped corneal opacity with extensive epithelial defects and nodular scleritis. Cysts of Acanthamoeba were identified in cultures from corneal scrapings, and he was diagnosed with ASK. He was started on antiamoebic treatment, including topical micafungin and chlorhexidine. The corneal ulcer was debrided several times. One month later, he developed necrotizing scleritis, and the cornea suddenly perforated. The eye was enucleated because of severe pain and prepared for histopathological examination. Results: The histopathological examination showed an infiltration of polymorphonuclear leukocytes throughout the corneal stroma and also in the limbal area of the sclera, forming an abscess. Granulation tissue was observed in the anterior sclera close to the ciliary body, but the posterior regions of the eye were not affected by inflammation or tissue destruction. The cysts of Acanthamoeba were observed only in the cornea. Conclusion: Histopathological examination of an eye with severe ASK showed that the inflammation and tissue granulation were present only in the anterior part of eye, and the posterior segment was not affected. Because the inflammation and tissue destruction were confined to the anterior segment, enucleation might not have been necessary if the severe pain was

  2. Tratamiento psicológico protocolizado en grupo de las enfermedades inflamatorias intestinales: efectos sobre la calidad de vida

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    Miguel Ángel Díaz-Sibaja

    2008-01-01

    Full Text Available La enfermedad inflamatoria intestinal (EII es una enfermedad crónica que afecta notablemente a la calidad de vida del paciente. El principal objetivo de este estudio fue demostrar la eficacia de un programa de tratamiento psicológico protocolizado para mejorar la calidad de vida de estos pacientes. La muestra estuvo constituida por 57 pacientes diagnosticados de EII (33 programa de tratamiento en grupo y 24 grupo control en lista de espera. Las variables dependientes fueron: síntomas digestivos, síntomas sistémicos, afectación funcional, función emocional y afectación social (Cuestionario de calidad de vida en la enfermedad inflamatoria intestinal (SIBDQ, López-Vivancos y cols., 1999. Los resultados revelan que el programa de tratamiento psicológico resultó eficaz para producir una mejoría significativa en todas las variables de calidad de vida a la finalización del mismo, al compararlo con el grupo control en lista de espera. Si bien las diferencias se mantienen en 4 de los 5 aspectos evaluados a los 3 meses de seguimiento, a los 12 sólo se mantienen las diferencias en uno de ellos.

  3. Quantitation by flow microfluorometry of total cellular DNA in Acanthamoeba

    Energy Technology Data Exchange (ETDEWEB)

    Coulson, P.B.; Tyndall, R.

    1978-01-01

    The DNA content of five speciea of Acanthamoeba was determined by flow microfluorometry. Acanthamoeba castellanii (AC-30), acanthamoeba polyphaga (APG and P-23), acanthamoeba rhysodes, acanthamoeba culbertsoni (A-1), and acanthamoeba royreba were grown in a casitone based medium 24 to 48 hr. The trophozoites were harvested, fixed in 70% ethanol (acidified), pretreated with RNase, stained with propidium diiodide, and evaluated for DNA-bound fluorescence. All species tested had DNA values between 2.0 to 5.0 pg/cell. These results placed DNA/cell values of Acanthamoeba slightly lower than DNA/cell values of other eucaryotic cells and much lower than Amoeba proteus values. These results indicate that FMF may be a useful adjunct in distinguishing Acanthamoeba cells from either eucaryotic cells or some other amoeba. However, differences in DNA/cell between species of Acanthamoeba are small and would not be useful in identification of species.

  4. Acanthamoeba keratitis after photorefractive keratectomy.

    Science.gov (United States)

    Kaldawy, Roger M; Sutphin, John E; Wagoner, Michael D

    2002-02-01

    A 37-year-old women developed severe suppurative keratitis immediately after having photorefractive keratectomy in her left eye. The keratitis was unresponsive to intensive topical antibiotic agents and topical and systemic steroids. Although the differential diagnosis included nonmicrobial and fungal keratitis, the clinical course and confocal microscopy suggested, and subsequent histopathologic examination confirmed, a diagnosis of Acanthamoeba keratitis. The amebic contamination probably resulted from exposure of the deepithelialized cornea to contaminated freshwater in a northern Wisconsin marsh. This case emphasizes the importance of encouraging patients with epithelial defects and bandage soft contact lenses to avoid exposure to contaminated freshwater until reepithelialization is complete.

  5. The immunological aspects of Acanthamoeba infections

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    Naveed A.  Khan

    2005-01-01

    Full Text Available Acanthamoeba is a protozoan pathogen that is responsible for two recognized diseases; i a blinding keratitis, most commonly associated with contact lens wear, and ii a rare but fatal granulomatous encephalitis that usually is limited to immunocompromised patients. The fact that Acanthamoeba infections are generally limited to avascular cornea or immunocompromised patients suggests that the normal immune responses may be sufficient to control and/or eradicate these pathogens. Here we describe our current understanding of the immune responses to Acanthamoeba infections.

  6. A comparative study of two histological techniques for the identification of cytomegalovirus infection in colorectal biopsies from patients with chronic inflammatory bowel disease Estudio comparativo entre dos técnicas histológicas para el diagnóstico de infección por citomegalovirus en biopsias colorrectales de pacientes con enfermedad inflamatoria intestinal crónica idiopática

    Directory of Open Access Journals (Sweden)

    M. L. de Castro

    2009-10-01

    Full Text Available Background: the role that cytomegalovirus (CMV plays in inflammatory bowel disease (IBD is controversial. The diagnosis of CMV infection in IBD depends on viral identification with hematoxylin-eosin (HE or immunohistochemistry (IHC. Our aim was to compare the sensitivity of HE and IHC for this diagnosis in IBD patients. Patients and methods: a case-control study. Our database was searched for IBD patients with HE- or IHC-based CMV-positivity from 1997 to 2007. Controls were selected among IBD inpatients matched for age and year of diagnosis with CMV. Their clinical characteristics were analyzed. HE and IHC were performed on biopsies from cases and controls at 6 months before and after inclusion in the study. In the statistical analysis, p values below 0.05 were considered significant. Results: ten IBD patients with CMV infection were identified. IBD-CMV patients were more steroid-resistant or steroid-dependent (p = 0.03, and underwent a higher number of colonic biopsies (p = 0.03. From 97 biopsies analyzed, 12 were HE-negative and IHC-positive, and 3 showed reversed results. The sensitivity of HE was 58.6%, 95% CI (38.9-78.3, and that of IHC was 89.7%, 95% CI (76.8-100. We did not find a good level of agreement between both techniques: kappa value 0.55, 95% CI (0.36-0.75. CMV positivity with IHC was associated with the use of more than one immunosuppressant drug, OR 13.5, 95% CI (1.2-152.2. Antiviral treatment was useful for CMV patients with steroid-dependent and steroid-refractory IBD. Conclusions: IHC shows a 30% higher sensitivity than HE for the diagnosis of CMV infection in IBD patients. There is no good level of agreement between both histological techniques.Introducción: el papel que el citomegalovirus (CMV juega en la enfermedad inflamatoria intestinal (EII es controvertido. El diagnóstico de infección por CMV en estos pacientes reside en su identificación en el tejido colónico mediante hematoxilina-eosina (HE o inmunohistoqu

  7. Acanthamoeba keratitis with Curvularia co-infection

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    Gupta N

    2010-01-01

    Full Text Available We report a case of Acanthamoeba keratitis with Curvularia co-infection. Acanthamoeba and fungal co-infection have been uncommonly reported in literature, worldwide. A classical history with a strong clinical suspicion and experienced laboratory personnel with systematic examination of corneal scrapings for bacterial, viral, parasitic and fungal causes are imperative for accurate diagnosis. Early diagnosis of Acanthamoeba keratitis or fungal infection followed by aggressive and appropriate treatment with effective agents is critical for the retention of good vision. Acanthamoeba keratitis is difficult to diagnose and, despite improvement in treatment options, may culminate in prolonged morbidity and significant loss of visual acuity. This case emphasizes the important role played by clinical microbiologists in making prompt diagnosis which can ultimately reduce visual morbidity.

  8. Enfermedad inflamatoria intestinal: situación actual de las alternativas terapéuticas

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    Lucrecia Suárez

    2007-03-01

    Full Text Available La enfermedad inflamatoria intestinal (EII se caracteriza por tener un curso clínico impredecible alternando recaídas con períodos de inactividad, una respuesta muy variable a la terapia y la constante aparición de complicaciones diversas. Su manejo es particularmente complejo en el niño y si bien existen diferentes guías y recomendaciones disponibles, los ensayos clínicos realizados en este grupo de edad son limitados y la evidencia científica es contradictoria, por lo cual aún no se cuenta con un consenso terapéutico aceptado internacionalmente. Los objetivos generales del tratamiento consisten en inducir y mantener la enfermedad en remisión el mayor tiempo posible, conseguir un adecuado crecimiento y prevenir posibles complicaciones. En años recientes, la aparición de nuevas alternativas terapéuticas ha permitido darle un enfoque más integral al manejo de estos pacientes tomando también en consideración el estado nutricional y psicológico además de su calidad de vida. Estos tratamientos, si bien consiguen en muchos casos controlar la inflamación intestinal curando las lesiones de la mucosa, no están exentos de efectos secundarios a corto y largo plazo, motivo por el cual es imprescindible tomar en cuenta las características de cada paciente para poder ofrecer un tratamiento individualizado que sea capaz de modificar la evolución natural de la enfermedad.

  9. Tratamiento nutricional en la enfermedad inflamatoria intestinal Nutritional management of inflammatory bowel disease

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    C. Pérez Tárrago

    2008-10-01

    Full Text Available Los pacientes con enfermedad inflamatoria intestinal presentan mayor riesgo de malnutrición. Por este motivo es muy frecuente que se precise un adecuado soporte nutricional. En estos pacientes se debe utilizar la nutrición enteral a menos que existan contraindicaciones. El soporte nutricional como tratamiento primario no está indicado en adultos (al haberse demostrado una mayor eficacia del tratamiento esteroideo salvo en caso de intolerancia o falta de respuesta al tratamiento médico. Por el contrario, la nutrición enteral se considera el tratamiento de primera línea en niños. No existe un claro beneficio con el uso de fórmulas específicas (grasa modificada, glutamina... por lo que no se aconseja su uso rutinario. A pesar de los grandes avances técnicos y científicos existen aún numerosos campos en los que ampliar conocimientos; algunos de los mismos se esbozan en la presente publicación.Patients with inflammatory bowel disease present higher risk for hyponutrition. For this reason, an adequate nutritional support is frequently needed. In these patients, enteral nutrition should be used unless there exist contraindications. Nutritional support as the primary therapy is no indicated in adults (since steroidal therapy has shown to be more effective but in the case of intolerance or lack of response to medical treatment. By contrast, enteral nutrition is considered a first line therapy in children. There is no clear benefit with the use of specific formulas (modified fat, glutamine..., so that their routine use is not recommended. In spite of the great technical and scientific advances, there are still many fields in which knowledge should be broaden; some of them are pointed out in this publication.

  10. Estudio PET/TC en patología inflamatoria-infecciosa PET/CT in infectious and inflammatory pathology

    OpenAIRE

    Cecilia Carrera; Silvina de Luca; Laura Tisser; Mariana Jakubowicz; Emilia Casalini Vañek; Eduardo Eyheremendy

    2012-01-01

    Objetivo. Demostrar la utilidad del examen PET/TC en patología inflamatoria-infecciosa. Materiales y Métodos. Evaluación retrospectiva (enero de 2009 - mayo de 2011) de los exámenes de tomografía por Emisión de Positrones/ Tomografía Computada (PET/TC), realizados en nuestra institución con un equipo híbrido SIEMENS-BIOGRAPH 16 (Siemens, Erlangen, Alemania). Se seleccionaron 5 pacientes. Resultados. Caso 1: paciente de 68 años de edad con fiebre de 6 meses de duración, fatiga y pérdida de pes...

  11. Contact Lens-Associated Acanthamoeba Keratitis in Iran

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    AH Maghsood

    2005-07-01

    Full Text Available Acanthamoeba keratitis is a vision-threatening infection caused by pathogenic species of the genus Acanthamoeba. In this study, 13 Acanthamoeba keratitis cases were diagnosed among 52 keratitis patients. To confirm the identity of Acanthamoeba at the genus level, a PCR-based method was used, and their pathogenic potential was determined using in vitro cytotoxicity assays on human corneal epithelial cells. Twelve (92.3% of Acanthamoeba keratitis patients were contact lens wearers; among them eleven (91.7% wore soft contact lenses. 11/13 (84.6% isolates were axenised in liquid culture medium, of which 10 (90.9% isolates disrupted corneal cells. Nine (69.2% isolates showed Acanthamoeba sp. group II, and four (30.8% showed group III morphology. To our knowledge this is the first report of determination of Acanthamoeba pathogenicity in Iran. This study confirms the importance of determination of pathogenic potential of Acanthamoeba isolates for clinical purposes.

  12. Eficacia histopatológica en la fase inflamatoria, proliferativa y osteogénica en la cicatrización alveolar post exodoncia aplicando matico (Piper angustifolium)

    OpenAIRE

    Pérez, Aurelia

    2016-01-01

    El presente estudio tiene como finalidad determinar la eficacia histopatológica en la fase inflamatoria, proliferativa y osteogénica de la cicatrización post exodoncia tratadas con Matico (Piper angustifolium). El estudio fue experimental, tuvo por finalidad saber el nivel de cicatrización histopatológica del Matico (Piper angustifolium) dentro de los alveolos post exodoncia, con un grupo de estudio de 15 conejos machos con un peso de 2.5 Kg, divididos en grupo control y grupo experimental (s...

  13. Variabilidad genética de la respuesta inflamatoria. I. Polimorfismo -511 C/T en el gen IL1β en diferentes subpoblaciones peruanas

    OpenAIRE

    Óscar Acosta; Luis Solano; Doris Huerta; Daniel Oré; José Sandoval; José Figueroa; Ricardo Fujita

    2012-01-01

    Anales de la Facultad de Medicina es una publicación de la Universidad Nacional Mayor de San Marcos. El polimorfismo -511 citosina/timina (-511 C/T) en la región promotora del gen interleuquina 1 beta (IL1β) está implicado en la producción diferencial de la citoquina y por tanto puede estar asociado a la respuesta inmuno-inflamatoria en obesidad, dislipidemias, cardiopatías, cáncer, infecciones, y el tratamiento con nutrientes y fármacos. Objetivos: Establecer la distribución de frec...

  14. Unexpected postmortem diagnosis of acanthamoeba meningoencephalitis in an immunocompetent child

    Science.gov (United States)

    Binesh, Fariba; Karimi, Mehran; Navabii, Hossein

    2011-01-01

    Meningoencephalitis caused by Acanthamoeba spp. is a rare opportunistic infection, difficult to diagnose and treat, which causes death in almost all cases. Here, the authors report a 5-year-old Iranian immunocompetent girl who died of fulminant acanthamoeba meningoencephalitis. To the authors’ knowledge, this is the first case of acanthamoeba meningoencephalitis in Iran. PMID:22679147

  15. Elastase secretion in Acanthamoeba polyphaga.

    Science.gov (United States)

    Ferreira, Gabriela A; Magliano, Ana C M; Pral, Elizabeth M F; Alfieri, Silvia C

    2009-11-01

    Acanthamoeba species are frequently isolated from soil and water collections. In the environment, the organisms multiply as phagotrophic trophozoites and encyst under adverse conditions. Several species are known to infect man, causing keratitis and opportunistic diseases. The mechanisms underlying tissue damage and invasion by the amoebae are being elucidated and the involvement of secreted peptidases, particularly serine peptidases, has been demonstrated. Here, elastase activity was examined in Acanthamoeba-conditioned medium (ACM), making use of elastin-Congo red (ECR) and synthetic peptide p-nitroanilide substrates. ACM hydrolysed ECR over a broad pH range and optimally at a pH of 7.5 and above. Indicating the activity of serine and metallopeptidases, Congo red release was potently inhibited by PMSF, antipain, chymostatin and 1,10-phenanthroline, partially reduced by elastatinal and EDTA, and unaffected by 1,7-phenanthroline and E-64. Screening with synthetic substrates mainly showed the activity of serine peptidases. ACM efficiently hydrolysed Suc-Ala(2)-Pro-Leu-pNA and Suc-Ala(2)-Pro-Phe-pNA over a broad pH range (7.0-9.5) and was weakly active against Suc-Ala(3)-pNA, a substrate found to be optimally hydrolysed at a pH around 7.0. Following ammonium sulfate precipitation of ACM proteins and FPLC analysis, the majority of the ECR-splitting activity, characterised as serine peptidases, bound to CM-sepharose and co-eluted with part of the Suc-Ala(2)-Pro-Phe-pNA-hydrolysing activity in a gradient of 0-0.6M NaCl. In the corresponding FPLC fractions, serine peptidases resolving in the region of 70-130kDa were detected in gelatin gels. Overall, the results demonstrate that trophozoites secrete elastases, and additionally suggest the high molecular weight serine peptidases as possible elastase candidates.

  16. Infliximab en pacientes con enfermedad ocular inflamatoria, refractarios a DARMES

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    Elmer R. García-Salazar

    2013-07-01

    Full Text Available Se describe la experiencia con infliximab (anticuerpo monoclonal con una potente acción antiinflamatoria en el tratamiento de enfermedades oculares inflamatorias secundarias a patologías reumáticas y refractarias a drogas antirreumáticas modificadoras de la enfermedad (DARMES. Se evaluó el caso de una paciente de 50 años con artritis reumatoide (AR de fondo activo y una paciente de 37 años con vasculitis anticuerpos anticitoplasma de neutrófilos especifico para mieloperoxidasa (ANCA MPO sin compromiso de órgano noble, ambas con escleritis bilateral y perforación con prolapso de iris del ojo izquierdo. Ellas recibieron infliximab EV en dosis de 3 a 5 mg/kg/dosis, según el esquema, a las 0, 2, 6 y 8 semanas. Infliximab resultó eficaz y seguro para el tratamiento de escleritis asociada a AR y vasculitis ANCA MPO positivo, refractaria a tratamiento con DARMES y corticoides en dosis altas. Los injertos de tejido esclerocorneal evolucionaron favorablemente con infliximab.

  17. FERTILIDAD Y EMBARAZO EN PACIENTES CON ENFERMEDADES INFLAMATORIAS INTESTINALES

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    ANDRÉS YARUR U., DR.

    2015-09-01

    Full Text Available Las Enfermedades Inflamatorias Intestinales (EII son entidades crónicas del tracto digestivo, que afectan frecuentemente a pacientes en edad reproductiva. Debido a las características de estas enfermedades y su tratamiento, existen múltiples desafíos. En este artículo, revisamos la evidencia más reciente con respecto a fertilidad y embarazo en pacientes con EII. En general, existe evidencia de que pacientes con EII tienen una mayor tasa de complicaciones durante el embarazo con respecto a pacientes sin EII. Sin embargo, esta diferencia está directamente asociada al grado de actividad de la enfermedad. La mayor parte de los fármacos hoy usados en el tratamiento de EII son considerados seguros durante el embarazo y se recomienda continuarlos, sobre todo considerando que el mayor riesgo de complicaciones está asociado a una enfermedad activa. Sin embargo es importante considerar las opciones caso a caso. Las dos grandes excepciones son metotrexato y talidomida que están completamente contraindicadas. La recomendación más importante es educar a toda paciente con EII en edad reproductiva, explicando que el embarazo debe llevarse a cabo cuando la enfermedad esté controlada y que la probabilidad de complicaciones está relacionada con el grado de actividad y severidad de la EII. Los médicos tratantes deben educar a las pacientes, enfatizando el seguimiento de los controles y tratamiento.

  18. Tigecycline inhibits proliferation of Acanthamoeba castellanii.

    Science.gov (United States)

    Jha, Bijay Kumar; Seo, Incheol; Kong, Hyun-Hee; Suh, Seong-Il; Suh, Min-Ho; Baek, Won-Ki

    2015-03-01

    Acanthamoeba is an opportunistic protozoan parasite responsible for different diseases in humans, such as granulomatous amoebic encephalitis and amoebic keratitis. Tigecycline, a third-generation tetracycline antibiotic, has potential activity to treat most of the antibiotic resistant bacterial infections. The effects of tigecycline in eukaryotic cells as well as parasites are less well studied. In the present study, we tested the effects of tigecycline on trophozoites of Acanthamoeba castellanii. The inhibitory effect of tigecycline on Acanthamoeba was determined by resazurin reduction and trypan blue exclusion assays. We found that tigecycline significantly inhibited the growth of Acanthamoeba (46.4 % inhibition at the concentration of 100 μM) without affecting cell viability and induction of encystation, whereas other tetracycline groups of antibiotics such as tetracycline and doxycycline showed no inhibitory effects. Furthermore, tigecycline decreased cellular adenosine triphosphate (ATP) level by 26 % than the control and increased mitochondrial mass, suggesting mitochondrial dysfunction in tigecycline-treated cells. These findings suggest that mitochondrial dysfunction with decreased ATP production might play an important mechanism of tigecycline in suppression of Acanthamoeba proliferation.

  19. A case of trauma related Acanthamoeba keratitis.

    Science.gov (United States)

    Kamel, A G M; Faridah, H; Yusof, S; Norazah, A; Nakisah, M A

    2004-12-01

    Acanthamoeba is an uncommon cause of keratitis but one of the most severe because of the prolonged and painful course of the disease and poor visual outcome. Although contact lens use is the principal risk factor, about 10% of cases occur following trauma and exposure to contaminated soil or water. Two cases of Acanthamoeba keratitis involving women contact lens wearers have previously been reported in Malaysia but this is the first time, a non contact lens related Acanthamoeba keratitis is reported. The case involved a 28 year old Indonesian male construction worker who had a trauma of the right eye during work. His eye was struck by sand and dust particles after which he quickly washed with water from an open tank at the construction site. He experienced pain, redness, glaring and blurring of vision of the right eye three days later. The diagnosis was missed at initial presentation but culture of the corneal scraping had proven Acanthamoeba as the aetiological agent. The history and clinical findings of this trauma related Acanthamoeba keratitis are briefly discussed.

  20. First cases of Acanthamoeba keratitis in Slovakia.

    Science.gov (United States)

    Ondriska, Frantisek; Mrva, Martin; Lichvár, Martin; Ziak, Peter; Murgasová, Zuzana; Nohýnková, Eva

    2004-01-01

    We present the case report of the first identification of Acanthamoeba as a causative agent of keratitis in the Slovak Republic. For the first time, Acanthamoeba sp. Group III was isolated from a 53-year-old patient with keratitis, which was manifested after an injury of the right eye. A delayed visit to a physician as well as a late diagnosis of the illness led to the advanced stage of eye disease. As the treatment with itraconazol and cornea transplantation showed no result, enucleation of the eye was decided. Acanthamoeba ludgunensis was also the causative agent of keratitis in a 39-year-old patient wearing contact lenses. His complaints occurred a month after bathing in a thermal swimming pool. The symptoms presented in the left eye were those of herpetic keratitis, and led to a cloudy cornea with circular infliltrate and poor vision. A prompt clinical and laboratory diagnosis, along with treatment with propamidine-isetionate resulted in a significant improvement of the eye condition. Contact lenses were probably related to another case of Acanthamoeba keratitis. The patient, a 15-year-old girl, kept wearing contact lenses during bathing in various swimming pools and in the sea; her contact lenses were also regularly washed under tap water. Due to the fact that cysts of Acanthamoeba sp. group II were found in the contact lens solution, this is presumed to be the source of the eye infection.

  1. Laboratory diagnosis of Acanthamoeba keratitis in Hungary.

    Science.gov (United States)

    Orosz, Erika; Farkas, Ágnes; Kucsera, István

    2016-09-01

    Acanthamoeba species are free-living amebae that can be found in almost every range of environments. Within this genus, numerous species are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK). AK is a corneal disease that is predominantly associated with contact lens use, the epidemiology of which is related to the specific genotype of Acanthamoeba. This study reports seven (7/16; 43.75%) positive cases. Detection of Acanthamoeba in corneal scrapings is based on cultivation and polymerase chain reaction (PCR) combined with the molecular taxonomic identification method. By PCR, seven samples were positive; cultivation was successful for five samples, probably because of the low quantity of samples. Genotype identification was carried out with a real-time fluorescence resonance energy transfer PCR assay based on sequence analysis of the 18S rRNA gene, and sensitivity and specificity were evaluated in comparison with traditional parasitological techniques. All seven detected Acanthamoeba strains belonged to the T4 genotype, the main AK-related genotype worldwide. These results confirmed the importance of a complete diagnostic protocol, including a PCR assay, for the clinical diagnosis of AK from human samples. Genotyping allowed the identification of all isolates in the T4 group, thus demonstrating the prevalence of this genotype in Hungary.

  2. Impacto de la nutrición en la evolución de la enfermedad inflamatoria intestinal Impact of nutritional treatment in the evolution of inflammatory bowel disease

    OpenAIRE

    M.ª D. Ballesteros Pomar; A. Vidal Casariego; A. Calleja Fernández; J. J. López Gómez; A. Urioste Fondo; I. Cano Rodríguez

    2010-01-01

    La enfermedad inflamatoria intestinal es una entidad de etiopatogenia aún no bien conocida, con importantes implicaciones nutricionales y metabólicas, por la alta prevalencia de malnutrición que conlleva; por la posible implicación de factores dietéticos en su patogenia; y por la hipótesis de que la intervención nutricional pudiera ser un tratamiento primario de la enfermedad. Algunos nutrientes, además de su función exclusivamente nutricional, podrían inducir un bajo estímulo antigénico, reg...

  3. Neuritis óptica inflamatoria Inflammatory optic neuritis

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    T. Ayuso

    2009-08-01

    Full Text Available La neuritis óptica inflamatoria (NO es la causa más frecuente de pérdida visual aguda en adultos jóvenes. Aunque el pronóstico visual es excelente en la mayoría de los casos, muchos pacientes desarrollarán otra patología como esclerosis múltiple en la evolución posterior. La historia natural de la NO ha sido estudiada en múltiples trabajos en los últimos años; uno de los más importantes es el Optic Neuritis Treatment Trial. La Resonancia Magnética tiene un papel fundamental en el diagnóstico etiológico de la NO y en la predicción del riesgo de conversión a esclerosis múltiple. Recientemente se han incorporado nuevas técnicas exploratorias como la tomografia de coherencia óptica, útil para el diagnóstico y pronóstico; se han identificado biomarcadores séricos que ayudan en el diagnóstico de otras patologías de naturaleza autoinmune que producen NO. Un mejor conocimiento de los datos clínicos y exploratorios de la NO típica permitirá un estudio diagnóstico más rápido y certero. El tratamiento de la NO con esteroides debe ser individualizado teniendo en cuenta que no modifican el pronóstico a largo plazo y en pacientes con alto riesgo de conversión a esclerosis múltiple debe plantearse terapia inmunomoduladora. Este trabajo revisa los datos existentes en la literatura referentes a las manifestaciones clínicas, el diagnóstico etiológico y diferencial y tratamiento de la NO inflamatoria.Inflammatory Optic Neuritis (ON is the most frequent cause of acute visual loss in young adults. Although the visual prognosis is excellent in the majority of cases, many patients develop pathology, such as multiple sclerosis, in its subsequent evolution. The natural history of ON has been studied in numerous works in recent years; one of the most important of which is Optic Neuritis Treatment Trial. Magnetic Resonance plays a fundamental role in the etiological diagnosis of ON and in predicting the risk of conversion into

  4. Hiperplasia Fibrosa Inflamatoria: reporte de un caso Inflammatory Fibrous Hyperplasia: case report

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    A Casian Romero

    2011-08-01

    Full Text Available Introducción: La Hiperplasia Fibrosa Inflamatoria es una alteración caracterizada por un agrandamiento del tejido conectivo; se considera una lesión reactiva ya que se presenta como respuesta a una irritante crónico local; es una lesión común en pacientes adolescentes y de edad adulta debido a que su etiología principal se asocia a maloclusión, al uso de aparatología protésica u ortodóntica mal ajustada, así como a la presencia de biofilm. Sin embargo, la población infantil, principalmente en dentición mixta, no está exenta de presentar esta alteración, aunque en menor proporción. Objetivo: Presentar el caso clínico de un paciente en dentición mixta que presenta una Hiperplasia Fibrosa Inflamatoria con el fin de presentar las características clínicas e histopatológicas de la lesión, enfatizando la importancia de un temprano y correcto diagnóstico. Presentación del Caso: Paciente masculino de 8 años de edad que se presenta a la clínica por presentar lesiones cariosas y malposición dental. A la exploración se observa un agrandamiento gingival localizado, en el órgano dentario 41. Se decide tomar una biopsia de la lesión para establecer el diagnóstico. Conclusión: Debido a que existen varias lesiones con características clínicas muy similares a las que presentaba el paciente, es necesario conocer los diagnósticos diferenciales, así como realizar el diagnóstico temprano de la lesión para poder llevar a cabo un tratamiento eficaz y adecuado.Introduction: The Inflammatory Fibrous Hyperplasia (IFH is a disorder consisting in a connective tissue overgrowth; it is considered a reactive lesion because it appears in response to a chronic and local irritant; this is a very common oral lesion in adolescents and adults, its etiology is associated with dental malocclusion, use of over-extended prosthetic and orthodontic appliances, and with the presence of biofilm (plaque. Nevertheless, child population, mainly patients

  5. Carbohydrate analysis of Acanthamoeba castellanii.

    Science.gov (United States)

    Dudley, Ricky; Jarroll, Edward L; Khan, Naveed Ahmed

    2009-08-01

    We analyzed biochemically Acanthamoeba castellanii trophozoites, intact cysts and cyst walls belonging to the T4 genotype using gas chromatography combined with mass spectrometry. Cyst walls were prepared by removing intracellular material from cysts by pre-treating them with sodium dodecyl sulphate (SDS) containing dithiothreitol, and then subjecting these to a series of sequential enzymatic digestions using amyloglucosidase, papain, DNase, RNase and proteinase K. The resulting "cyst wall" material was subsequently lyophilized and subjected to glycosyl composition analysis. Transmission electron microscopy confirmed the removal of intracystic material following enzymatic treatment. Our results showed that treated A. castellanii trophozoites, intact cysts and cyst walls contained various sugar moieties, of which a high percentage was galactose and glucose, in addition to small amounts of mannose, and xylose. Linkage analysis revealed several types of glycosidic linkages including the 1,4-linked glucosyl conformation, indicative of cellulose. Inhibitor studies suggested that, beside sugar synthesis, cytoskeletal re-arrangement and mitogen-activated protein kinase-mediated pathways are involved in A. castellanii encystment.

  6. La expresión de IL-10 interviene en la regulación de la respuesta inflamatoria por los ácidos grasos omega 3 IL-10 expression is involved in the regulation of the immune response by omega 3 fatty acids

    Directory of Open Access Journals (Sweden)

    S. Sierra

    2004-11-01

    Full Text Available Objetivo o antecedente: Los ácidos grasos poliinsaturados son importantes para el organismo humano debido a su implicación en numerosas funciones biológicas. Las dietas occidentales se caracterizan por ser excesivamente ricas en ácidos grasos w-6 y pobres en ácidos grasos w-3. Los ácidos grasos de la serie w-3 son necesarios para el normal crecimiento y desarrollo del individuo así como para la regulación de la respuesta inmunológica. El objetivo de este estudio es analizar el efecto de una dieta enriquecida en ácidos grasos w-3 frente a un proceso inflamatorio así como el estudio de los mecanismos implicados en dicho efecto. Intervenciones: Para ello, ratones Balb/c fueron alimentados durante un mes con una dieta cuya fuente lipídica era 100% aceite de girasol (control, o con la misma dieta en la que el 12% de la grasa era aceite de pescado y el resto aceite de girasol (W-3. Doce horas antes de su sacrificio se indujo en una de las orejas de cada animal una dermatitis de contacto que cursó con inflamación y edema. Como agente inflamatorio se utilizó 2,4 dinitrofluorobenceno. Tras el sacrificio se tomaron diversas muestras y se analizaron. Resultados: La inflamación, medida como peso y contenido de agua de las orejas, disminuyó significativamente en los ratones alimentados con w-3. La medida de la infiltración leucocitaria y los parámetros de oxidación revelaron también la mejora en el proceso inflamatorio de dichos ra-tones. Para explicar estos hechos se analizó la expresión de diversas citocinas, observándose un incremento de IL-10 y una disminución de citocinas tanto Th1 como Th2. Conclusiones: Los ácidos grasos w-3 poseen un efecto inmunomodulador al actuar como antiinflamatorios y antialérgicos, al tiempo que aumentan algunas defensas del organismo. La citocina reguladora IL-10 podría ser la responsable del efecto antiinflamatorio ejercido por los ácidos grasos w-3.Introduction: Polyunsaturated fatty acids play

  7. Rol de galectina-1 en la fisiopatología de las Enfermedades Inflamatorias Intestinales

    OpenAIRE

    Papa Gobbi, Rodrigo

    2015-01-01

    Las Enfermedades Inflamatorias Intestinales (EII) son un conjunto de patologías crónicas multifactoriales en las cuales se produce una respuesta inmune aberrante contra componentes de la microbiota intestinal. Dentro de las EII, la colitis ulcerosa (CU) y la enfermedad de Crohn (EC) son las dos formas más estudiadas. Si bien existen diferencias en la presentación clínica de estas patologías, en ambos casos los linfocitos T (LT) de la mucosa sobre-expresan citoquinas pro-inflamatorias del perf...

  8. Respuesta inflamatoria esplácnica en la colestasis microquirúrgica experimental

    OpenAIRE

    García Domínguez, José

    2008-01-01

    La colestasis extrahepática en la rata cursa con alteraciones esplácnicas y sistémicas en cuya producción podrían estar implicados mecanismos patogénicos de naturaleza inflamatoria. Con el objeto de caracterizar el tipo de respuesta inflamatoria esplácnica secundaria a la colestasis obstructiva, se utilizó un modelo experimental de colestasis extrahepática microquirúrgica, para estudiar en una fase evolutiva considerada crónica, la concentración de citoquinas tipo Th1 (TNFα e IL-1) y Th2 (...

  9. Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis.

    Science.gov (United States)

    Booton, Gregory C; Rogerson, Andrew; Bonilla, Tonya D; Seal, David V; Kelly, Daryl J; Beattie, Tara K; Tomlinson, Alan; Lares-Villa, Fernando; Fuerst, Paul A; Byers, Thomas J

    2004-01-01

    Previous molecular examination of Acanthamoeba spp. has resulted in the determination of distinct genotypes in this genus (designated T1-T12, T14). Genotype T4 has been responsible for the majority of cases of Acanthamoeba keratitis. Here we examine the relative abundance of environmental T4 isolates on beaches and ask whether they have temperature and salinity tolerances that could enhance pathogenicity. Twenty-four Acanthamoeba strains were isolated from beach sand (n = 20), soil (n = 3), and tap water (n = 1) in south Florida. Phylogenetic analysis identified 19 of 24 isolates as T4, the Acanthamoeba keratitis-associated genotype. The remaining isolates were genotype T5 (4) and T11 (1). Nearly all beach isolates were genotype T4, whereas the tap water and soil isolates were mostly T5. All amoebae grew at 0, 1.0, and 2.0% salt and 19 of 20 beach isolates also grew at 3.2%. No soil or tap-water acanthamoebae reproduced at 3.2%. All isolates grew at 37 degrees C and two (T5) at 42 degrees C. Little correlation existed between beach location, salt-tolerance, and genetic relatedness. Overall, the large majority of environmental isolates obtained were genotype T4, suggesting it may be the most common genotype in this environment and could be a potential source of Acanthamoeba keratitis infections.

  10. Autophagy inhibitors as a potential antiamoebic treatment for Acanthamoeba keratitis.

    Science.gov (United States)

    Moon, Eun-Kyung; Kim, So-Hee; Hong, Yeonchul; Chung, Dong-Il; Goo, Youn-Kyoung; Kong, Hyun-Hee

    2015-07-01

    Acanthamoeba cysts are resistant to extreme physical and chemical conditions. Autophagy is an essential pathway for encystation of Acanthamoeba cells. To evaluate the possibility of an autophagic Acanthamoeba encystation mechanism, we evaluated autophagy inhibitors, such as 3-methyladenine (3MA), LY294002, wortmannin, bafilomycin A, and chloroquine. Among these autophagy inhibitors, the use of 3MA and chloroquine showed a significant reduction in the encystation ratio in Acanthamoeba cells. Wortmannin also inhibited the formation of mature cysts, while LY294002 and bafilomycin A did not affect the encystation of Acanthamoeba cells. Transmission electron microscopy revealed that 3MA and wortmannin inhibited autophagy formation and that chloroquine interfered with the formation of autolysosomes. Inhibition of autophagy or autolysosome formation resulted in a significant block in the encystation in Acanthamoeba cells. Clinical treatment with 0.02% polyhexamethylene biguanide (PHMB) showed high cytopathic effects on Acanthamoeba trophozoites and cysts; however, it also revealed high cytopathic effects on human corneal epithelial cells. In this study, we investigated effects of the combination of a low (0.00125%) concentration of PHMB with each of the autophagy inhibitors 3MA, wortmannin, and chloroquine on Acanthamoeba and human corneal epithelial cells. These new combination treatments showed low cytopathic effects on human corneal cells and high cytopathic effects on Acanthamoeba cells. Taken together, these results provide fundamental information for optimizing the treatment of Acanthamoeba keratitis.

  11. Twenty years of acanthamoeba diagnostics in Austria.

    Science.gov (United States)

    Walochnik, Julia; Scheikl, Ute; Haller-Schober, Eva-Maria

    2015-01-01

    Acanthamoebae are the causative agents of an often seriously progressing keratitis (AK) occurring predominantly in contact lens wearers and can cause several disseminating infections potentially resulting in granulomatous amoebic encephalitis (GAE) in the immunocompromised host. Our institution is the Austrian reference laboratory for Acanthamoeba diagnostics and the aim of this study was to give an overview of proven cases of Acanthamoeba infections in Austria during the past 20 yr. All samples of patients with suspected AK or GAE were screened for Acanthamoeba spp. by culture and/or PCR and the detected amoebae were genotyped. Altogether, 154 cases of AK and three cases of GAE were diagnosed. Age of the AK patients ranged from 8 to 82 yr (mean 37.8) and 58% of the patients were female. Approximately 89% of the AK patients were contact lens wearers, almost all cases were unilateral and 19% of the patients required a keratoplasty. Age of the GAE patients ranged from 2 to 25 yr (mean 14.7), all were HIV-negative, but two were severely immunosuppressed at the time of diagnosis. The predominant genotype in the AK cases was T4, other genotypes found were T3, T5, T6, T10 and T11. The three GAE cases involved genotypes T2, T4 and T5.

  12. The Value of Cytology Smears for Acanthamoeba Keratitis

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    Sangita P. Patel

    2016-01-01

    Full Text Available Purpose. Acanthamoeba keratitis remains a difficult diagnosis despite advances in genetic and imaging technologies. The purpose of this paper is to highlight the utility of cytology smears for diagnosis of Acanthamoeba keratitis. Methods. This is a case study of the diagnostic course for a patient with suspected Acanthamoeba keratitis. Results. A 40-year-old male with poor contact lens hygiene presented with severe left eye pain. Slit lamp examination showed two peripheral ring infiltrates without an epithelial defect. The epithelium over both infiltrates was removed with a Kimura spatula. Half of the sample was smeared on a dry microscope slide and the other half was submitted for Acanthamoeba culture and PCR. Both culture and PCR were negative for Acanthamoeba, but hematoxylin and eosin stain of the smear revealed double-walled cysts. Conclusion. H&E staining of corneal cytology specimens is an efficient and readily available test for diagnosis of Acanthamoeba keratitis.

  13. The Value of Cytology Smears for Acanthamoeba Keratitis

    Science.gov (United States)

    Schaefer, Jamie L.; Paterson, Joyce; Liu, Weiguo; Gonzalez-Fernandez, Federico

    2016-01-01

    Purpose. Acanthamoeba keratitis remains a difficult diagnosis despite advances in genetic and imaging technologies. The purpose of this paper is to highlight the utility of cytology smears for diagnosis of Acanthamoeba keratitis. Methods. This is a case study of the diagnostic course for a patient with suspected Acanthamoeba keratitis. Results. A 40-year-old male with poor contact lens hygiene presented with severe left eye pain. Slit lamp examination showed two peripheral ring infiltrates without an epithelial defect. The epithelium over both infiltrates was removed with a Kimura spatula. Half of the sample was smeared on a dry microscope slide and the other half was submitted for Acanthamoeba culture and PCR. Both culture and PCR were negative for Acanthamoeba, but hematoxylin and eosin stain of the smear revealed double-walled cysts. Conclusion. H&E staining of corneal cytology specimens is an efficient and readily available test for diagnosis of Acanthamoeba keratitis. PMID:27403362

  14. Enfermedad inflamatoria intestinal en pacientes celíacos Inflammatory bowel disease in celiac patients

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    M. Masachs

    2007-08-01

    Full Text Available Introducción: se ha sugerido una potencial asociación entre la enfermedad celíaca y la enfermedad inflamatoria intestinal, que puede justificar que ambas enfermedades puedan presentarse en un mismo enfermo o en sus familiares de primer orden con mayor frecuencia de lo esperado. Objetivo: determinar la prevalencia de la enfermedad de Crohn y la colitis ulcerosa en los enfermos celíacos y en sus familiares. Método: estudio epidemiológico prospectivo transversal en un grupo de pacientes celíacos, sus familiares de primer grado y un grupo control de características epidemiológicas similares, constituido por familiares de pacientes que acuden al Servicio de Urgencias por un problema agudo. Para detectar la existencia de colitis ulcerosa y enfermedad de Crohn en los celíacos y sus familiares, se realizó una entrevista semiestructurada. Resultados: se han incluido 86 celíacos y 432 familiares, que se han comparado con 809 controles (129 pacientes con una enfermedad aguda y 680 familiares de primer grado suyos. Se han detectado 3 casos de enfermedad de Crohn en el grupo de los enfermos celíacos y 4 casos de enfermedad de Crohn en sus familiares. Sólo se ha detectado 1 caso de enfermedad de Crohn en el grupo control (p Introduction: a potential association between celic disease and inflammatory bowel disease hs been suggested, which may explain the fact that both disorders occasionally present in one patient or in his/her first-degree relatives more frequently than expected. Objective: to establish the prevalence of Crohn's disease and ulcerative colitis in celiac patients and their relatives. Method: a cross-sectional, prospective epidemiological study in a group of celiac patients, their first-degree relatives, and a control group with similar epidemiological characteristics including the relatives of patients presenting at the ER for acute conditions. A semistructured interview was used to identify the presence of Crohn's disease and

  15. Principales protagonistas de la respuesta inflamatoria a la infección

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    Juan Carlos Velázquez Acosta

    1998-06-01

    Full Text Available Se hace una revisión de los principales elementos implicados en la respuesta inflamatoria del individuo a la infección, por el beneficio que este conocimiento reporta al mejor manejo terapéutico de los pacientes. Se revisan los principales protagonistas de la inflamación, desde los detonantes como: lipopolisacárido, peptidoglicano, ácidos lipoteicoico y murámico, hasta los principales mediadores implicados. Se discute el proceso de activación de monocitos y células endoteliales y la repercusión de ello, y se hace un análisis dinámico del proceso de activación en función del tiempo trancurrido desde el comienzo de la infección, y sus etapas fisiopatológicas. Se evidencia el papel protagónico que tiene el propio organismo en el daño producido por la respuesta a la infección, además de la importancia del adecuado equilibrio entre proinflamación y contrarregulación para el pronóstico del enfermo, lo que obliga a tenerlo en cuenta a la hora del tratamiento.A review of the main elements involved in the inflamatory response of the individuals to infection is made, taking into account that this knowledge helps to improve the therapeutic management of patients. The main agents of inflammation are reviewed, starting from detonators such as: lipopolysaccharides, petpidoglycans, muramic acid to the principal mediators involved. The activation process of monocytes and endothelial cells is dicussed, as well as its repercussion. A dynamic analysis of the activation process according to the time elapsed since the onset of the infection and of its physiopathological stages is made. It is demonstrated the fundamental role played by the organism in the damage produced by the response to infection, and the importance of an adequate balance between proinflammation and contraregulation for the patient´s prognosis, which makes it necessary at the time of treatment.

  16. A clinical Acanthamoeba isolate harboring two distinct bacterial endosymbionts.

    Science.gov (United States)

    Müller, Anneliese; Walochnik, Julia; Wagner, Martin; Schmitz-Esser, Stephan

    2016-10-01

    Acanthamoebae feed on bacteria but are also frequent hosts of bacterial symbionts. Here, we describe the stable co-occurrence of two symbionts, one affiliated to the genus Parachlamydia and the other to the candidate genus Paracaedibacter (Alphaproteobacteria), within a clinical isolate of Acanthamoeba hatchetti genotype T4. We performed fluorescence in situ hybridization (FISH) and transmission electron microscopy (TEM) to describe this symbiosis. Our study adds to other reports of simultaneous co-occurrence of two symbionts within one Acanthamoeba cell.

  17. Acanthamoeba: biology and increasing importance in human health.

    Science.gov (United States)

    Khan, Naveed Ahmed

    2006-07-01

    Acanthamoeba is an opportunistic protozoan that is widely distributed in the environment and is well recognized to produce serious human infections, including a blinding keratitis and a fatal encephalitis. This review presents our current understanding of the burden of Acanthamoeba infections on human health, their pathogenesis and pathophysiology, and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be targets for therapeutic interventions and/or the development of preventative measures.

  18. Acanthamoeba species in Swimming Pools of Cairo, Egypt.

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    Ahmad Al-Herrawy

    2014-06-01

    Full Text Available The free-living amoebae Acanthamoeba spp. have been recognized as etiologic agents of amoebic encephalitis, keratitis, otitis, lung lesions and other skin infections mainly in immuno-compromised individuals. The purpose of this study is to detect the presence of Acanthamoeba in swimming pools in Egypt using a polymerase chain reaction (PCR method.Water samples were collected from 10 different swimming pools in Cairo, Egypt. Samples were cultured on non-nutrient agar for the detection of Acanthamoeba isolates that were confirmed by PCR amplification using genus specific primers. The molecularly confirmed Acanthamoeba isolates were morphologically identified to the species level.Members of genus Acanthamoeba were detected in 49.2% of the examined swimming-pool water samples. Morphologically, six Acanthamoeba species were isolated from the examined swimming pool water namely A. polyphaga, A.castellanii, A. rhysodes, A. mauritaniensis, A. royreba and A. triangularis. All the identified species of Acanthamoeba were molecularly confirmed to be related to the genus Acanthamoeba.The isolated species of Acanthamoeba could provoke variable degrees of infections to the swimmers. The culture method is cheaper and easier than PCR techniques that are faster for the detection of free-living amoebae.

  19. Area 51: How do Acanthamoeba invade the central nervous system?

    Science.gov (United States)

    Siddiqui, Ruqaiyyah; Emes, Richard; Elsheikha, Hany; Khan, Naveed Ahmed

    2011-05-01

    Acanthamoeba granulomatous encephalitis generally develops as a result of haematogenous spread, but it is unclear how circulating amoebae enter the central nervous system (CNS) and cause inflammation. At present, the mechanisms which Acanthamoeba use to invade this incredibly well-protected area of the CNS and produce infection are not well understood. In this paper, we propose two key virulence factors: mannose-binding protein and extracellular serine proteases as key players in Acanthamoeba traversal of the blood-brain barrier leading to neuronal injury. Both molecules should provide excellent opportunities as potential targets in the rational development of therapeutic interventions against Acanthamoeba encephalitis.

  20. Identification of Paenibacillus as a Symbiont in Acanthamoeba.

    Science.gov (United States)

    Maschio, Vinicius José; Corção, Gertrudes; Bücker, Francielle; Caumo, Karin; Rott, Marilise Brittes

    2015-09-01

    Amoebae of the genus Acanthamoeba occur worldwide and in addition to being pathogens, are important vehicles for microorganisms with clinical and environmental importance. This study aimed to evaluate the profiling of endosymbionts in 12 isolates of Acanthamoeba using V3 region of 16S rDNA denaturing gradient gel electrophoresis (DGGE) and sequencing. The DGGE enabled us to characterize the endosymbionts diversity in isolates of Acanthamoeba, and to identify Paenibacillus sp., an emerging pathogen, as an amoebic endosymbiont. The results of this study demonstrated that Acanthamoeba is capable of transporting a large number of endosymbionts. This is the first study that reports, the presence of Paenibacillus sp. as amebic symbiont.

  1. Is there evidence of sexual reproduction (meiosis) in Acanthamoeba?

    Science.gov (United States)

    Khan, Naveed A; Siddiqui, Ruqaiyyah

    2015-06-01

    Evolution of independently breeding species into males and females (gametes) has remained a puzzle. Given the significant advantages of sexual reproduction over asexual reproduction as a long-term species survival strategy; here, we pose the question whether there is some form of meiosis in Acanthamoeba species, which represents our ancient lineage. The recently available Acanthamoeba genome revealed several genes implicated in meiosis in sexual eukaryotes such as Spo11, Mre11, Rad50, Rad51, Rad52, Mnd1, Dmc1, Msh, and Mlh, suggesting that Acanthamoeba is capable of some form of meiosis, inferring the presence of sexual reproduction in Acanthamoeba, and that meiosis evolved early in eukaryotic evolution.

  2. Acanthamoeba keratitis: improving the Scottish diagnostic service for the rapid molecular detection of Acanthamoeba species.

    Science.gov (United States)

    Alexander, Claire Low; Coyne, Michael; Jones, Brian; Anijeet, Deepa

    2015-07-01

    Acanthamoeba species are responsible for causing the potentially sight-threatening condition, Acanthamoeba keratitis, which is commonly associated with contact lens use. In this report, we highlight the challenges faced using conventional laboratory identification methods to identify this often under-reported pathogen, and discuss the reasons for introducing the first national service in Scotland for the rapid and sensitive molecular identification of Acanthamoeba species. By comparing culture and molecular testing data from a total of 63 patients (n = 80 samples) throughout Scotland presenting with ocular eye disease, we describe the improvement in detection rates where an additional four positive cases were identified using a molecular assay versus culture. The testing of a further ten patients by confocal imaging is also presented. This report emphasizes the importance of continuing to improve clinical laboratory services to ensure a prompt, correct diagnosis and better prognosis, in addition to raising awareness of this potentially debilitating opportunistic pathogen.

  3. The Development of Drugs against Acanthamoeba Infections.

    Science.gov (United States)

    Siddiqui, Ruqaiyyah; Aqeel, Yousuf; Khan, Naveed Ahmed

    2016-11-01

    For the past several decades, there has been little improvement in the morbidity and mortality associated with Acanthamoeba keratitis and Acanthamoeba encephalitis, respectively. The discovery of a plethora of antiacanthamoebic compounds has not yielded effective marketed chemotherapeutics. The rate of development of novel antiacanthamoebic chemotherapies of translational value and the lack of interest of the pharmaceutical industry in developing such chemotherapies have been disappointing. On the other hand, the market for contact lenses/contact lens disinfectants is a multi-billion-dollar industry and has been successful and profitable. A better understanding of drugs, their targets, and mechanisms of action will facilitate the development of more-effective chemotherapies. Here, we review the progress toward phenotypic drug discovery, emphasizing the shortcomings of useable therapies. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  4. Acanthamoeba keratitis: report of three cases.

    Science.gov (United States)

    Maudgal, P C

    1989-01-01

    During the last year we diagnosed three patients to have Acanthamoeba keratitis. In one patient, who wore soft contact lenses, keratitis had developed after swimming in a contaminated heated swimming pool. Neither a history of trauma was present in two other patients nor did they wear contact lenses. Diagnosis of Acanthamoeba was established by Calcofluor White (cellufluor) staining of the scraped corneal material. Patients were treated with Flagyl (metronidazole) 0.5% eyedrops, Brolene (propamidine isethionate) 0.1% eyedrops, Neomycin eyedrops, topical corticosteroids (including sustain-release corticosteroid subconjunctival injections) and mydriatics. Keratitis in one patient healed within one month with full visual recovery, whereas the disease nearly ameliorated in the second patient. Corneal inflammation subsided markedly in the third patient with medical therapy, but a keratoplasty had to be performed because of a central opaque and ectatic cornea. This patient had already developed severe keratitis with necrosis of the cornea at the time of diagnosis.

  5. Influence of Acanthamoeba genotype on clinical course and outcomes for patients with Acanthamoeba keratitis in Spain.

    Science.gov (United States)

    Arnalich-Montiel, Francisco; Lumbreras-Fernández, Blanca; Martín-Navarro, Carmen M; Valladares, Basilio; Lopez-Velez, Rogelio; Morcillo-Laiz, Rafael; Lorenzo-Morales, Jacob

    2014-04-01

    Genotype T4 is by far the most frequent genotype of Acanthamoeba keratitis (AK) and therefore has been considered the most virulent. This study included 14 cases of AK of genotype T4 and three cases of non-T4 genotype. We found that cases of non-T4 genotype had a worse response to medical therapy, greater need for surgical intervention, greater risk of extracorneal involvement, and remarkably poorer final visual outcome than those of T4 genotype, suggesting an association between Acanthamoeba virulence and genotype that requires additional case investigation.

  6. Phagocytosis Affects Biguanide Sensitivity of Acanthamoeba spp.

    Science.gov (United States)

    Noble, Judith A.; Ahearn, Donald G.; Avery, Simon V.; Crow Jr., Sidney A.

    2002-01-01

    The incidence of Acanthamoeba keratitis, a disease associated with contact lens wear, has been in apparent decline with the advent of multipurpose contact lens solutions. The concentrations of the biguanides chlorhexidine digluconate (CHX) and particularly polyhexamethylene biguanide (PHMB) included in multipurpose solutions (MPSs) are sublethal for amoebae. We evaluated by flow cytometry the effects of these two biguanides on phagocytosis of particles and the survival of trophozoites of Acanthamoeba castellanii and A. polyphaga. Trophozoites of A. castellanii and A. polyphaga (106/ml) were exposed to solutions of 5 and 50 μg of PHMB and CHX per ml in the presence and absence of particles (i.e., heat-killed yeasts and bacteria and latex beads). In addition, trophozoites were exposed to particles treated with these concentrations of the two biguanides. In the absence of particles, trophozoites of A. polyphaga appeared to be more resistant to the biguanides than those of A. castellanii. In the presence of particles, the rates of survival of both species were decreased. In most instances, particles treated with sublethal concentrations of both biguanides that were adsorbed onto the particles reduced the incidence of phagocytosis. Particles present in MPSs in contact lens cases may be involved in the decreased incidence of Acanthamoeba keratitis. PMID:12069957

  7. Phagocytosis affects biguanide sensitivity of Acanthamoeba spp.

    Science.gov (United States)

    Noble, Judith A; Ahearn, Donald G; Avery, Simon V; Crow, Sidney A

    2002-07-01

    The incidence of Acanthamoeba keratitis, a disease associated with contact lens wear, has been in apparent decline with the advent of multipurpose contact lens solutions. The concentrations of the biguanides chlorhexidine digluconate (CHX) and particularly polyhexamethylene biguanide (PHMB) included in multipurpose solutions (MPSs) are sublethal for amoebae. We evaluated by flow cytometry the effects of these two biguanides on phagocytosis of particles and the survival of trophozoites of Acanthamoeba castellanii and A. polyphaga. Trophozoites of A. castellanii and A. polyphaga (10(6)/ml) were exposed to solutions of 5 and 50 microg of PHMB and CHX per ml in the presence and absence of particles (i.e., heat-killed yeasts and bacteria and latex beads). In addition, trophozoites were exposed to particles treated with these concentrations of the two biguanides. In the absence of particles, trophozoites of A. polyphaga appeared to be more resistant to the biguanides than those of A. castellanii. In the presence of particles, the rates of survival of both species were decreased. In most instances, particles treated with sublethal concentrations of both biguanides that were adsorbed onto the particles reduced the incidence of phagocytosis. Particles present in MPSs in contact lens cases may be involved in the decreased incidence of Acanthamoeba keratitis.

  8. Contact lens-related acanthamoeba keratitis.

    Science.gov (United States)

    Stapleton, Fiona; Ozkan, Jerome; Jalbert, Isabelle; Holden, Brien A; Petsoglou, Con; McClellan, Kathy

    2009-10-01

    Acanthamoeba keratitis is a rare but severe disease, with more than 95% of cases occurring in contact lens wearers. With a worldwide resurgence of contact lens-related disease, this report illustrates the clinical characteristics and treatment challenges representative of this disease. This report describes Acanthamoeba keratitis in a 47-year-old female using extended wear silicone hydrogel contact lenses, with a history of swimming in a home pool and failure to subsequently disinfect the contact lenses. The diagnosis was based on clinical signs, disease course, and confocal microscopy results despite a negative result for corneal smear and culture. The corneal signs included an epithelial defect, epithelial irregularities, anterior stromal infiltrates, perineural infiltrates, an anterior stromal ring infiltrate, and hypopyon. The case was diagnosed as an infective keratitis and treated promptly using intensive topical administration of fortified gentamicin and cephalothin. The high likelihood Acanthamoeba prompted immediate use of polyhexamethylbiguanide and chlorhexidine, with propamide and adjunct treatment using atropine and oral diclofenac. Steroids were added on day 3, and the frequency of administration of antibacterial treatment was gradually reduced and ceased by day 10. The analgesia was stopped at 3 months. The frequency of administration of antiamoeba therapy and steroid treatment was slowly reduced and all treatment was ceased after 18 months. Despite considerable morbidity in terms of the treatment duration, hospitalization, outpatient appointments, and associated disease costs, the final visual outcome (6/6) was excellent.

  9. Current Status of Acanthamoeba in Iran: A Narrative Review Article.

    Directory of Open Access Journals (Sweden)

    Maryam Niyyati

    2015-06-01

    Full Text Available Free-living amoebae belonging to the genus Acanthamoeba have an environmental distribution. Amoebic keratitis due to these protozoan parasites continue to rise in Iran and worldwide. In Iran, there are various researches regarding both morphological and molecular identification of Acanthamoeba spp. in environmental and clinical samples. However, there is no thorough review about Acanthamoeba genotypes and their distribution in environmental sources such as water, dust and biofilm in Iran. Besides, according to increasing cases of Amoebic keratitis in the region awareness regarding the pathogenic potential of these sight-threatening amoebae is of utmost importance.We conducted a thorough review based on the database sources such as MEDLINE, PubMed and Google scholar. No restrictions were placed on study date, study design or language of publication. We searched all valuable and relevant information considering the occurrence of the Acanthamoeba in both environmental and clinical samples.According to our thorough review Acanthamoeba belonging to T4 genotype is the most prevalent type strain in environmental and clinical samples in several regions in Iran and worldwide, however, there are reports regarding Acanthamoeba belonging to other genotypes such as T2, T3, T5, T6 and T11 and the mentioned point could leads us to more researches with the goal of presenting the real genotype dominance of Acanthamoeba and related disease in the country.Overall, the present review will focus on present status of genotypes of Acanthamoeba in Iran during recent years.

  10. Cytotoxic effect of acriflavine against clinical isolates of Acanthamoeba spp.

    Science.gov (United States)

    Polat, Zubeyda Akin; Karakus, Gulderen

    2013-02-01

    Acanthamoeba keratitis (AK) is a potentially devastating and sight-threatening infection of the cornea caused by the ubiquitous free-living amoebae, Acanthamoeba species. Its eradication is difficult because the amoebas encyst, making it highly resistant to anti-amoebic drugs. Acriflavine neutral (ACF) has been used for treatment of microbial infections for humans and fishes. The aim of our study was to evaluate the time-dependent cytotoxicities of ACF against Acanthamoeba spp. Trophozoites and cysts of three different strains (strain PAT06 Acanthamoeba castellanii, strain 2HH Acanthamoeba hatchetti, and strain 11DS A. hatchetti) of Acanthamoeba spp. were tested. All strains had been isolated from patients suffering from a severe AK. The effects of the ACF with the concentrations ranging from 15 to 500 mg mL(-1) on the cytotoxicity of Acanthamoeba strains were examined. ACF showed a time- and dose-dependent amebicidal action on the trophozoites and cysts. Pat06 (A. castellanii) was the most resistant, while strain 11DS (A. hatchetti) was the most sensitive. As a result, ACF could be concluded as a new agent for the treatment of Acanthamoeba infections. On the other hand, it still needs to be further evaluated by in vivo test systems to confirm the efficiency of its biological effect.

  11. Modulación de la respuesta inflamatoria esplácnica en la colestasis microquirúrgica experimental con ketotifén

    OpenAIRE

    Cruz Cidoncha, Arturo

    2013-01-01

    "Introducción". El desarrollo de fibrosis hepática e hipertensión portal en múltiples hepatopatías crónicas de etiología diversa es un proceso multifuncional, mediado por células inflamatorias que, al liberar mediadores fibrogénicos, activan a las células estrelladas y a los miofibroblastos. La administración de ketotifén, un fármaco estabilizador de células cebadas, que inhibe la síntesis y liberación de sus mediadores inflamatorios, en un modelo experimental microquirúrgico de colesta...

  12. Acanthamoeba induces cell-cycle arrest in host cells.

    Science.gov (United States)

    Sissons, James; Alsam, Selwa; Jayasekera, Samantha; Kim, Kwang Sik; Stins, Monique; Khan, Naveed Ahmed

    2004-08-01

    Acanthamoeba can cause fatal granulomatous amoebic encephalitis (GAE) and eye keratitis. However, the pathogenesis and pathophysiology of these emerging diseases remain unclear. In this study, the effects of Acanthamoeba on the host cell cycle using human brain microvascular endothelial cells (HBMEC) and human corneal epithelial cells (HCEC) were determined. Two isolates of Acanthamoeba belonging to the T1 genotype (GAE isolate) and T4 genotype (keratitis isolate) were used, which showed severe cytotoxicity on HBMEC and HCEC, respectively. No tissue specificity was observed in their ability to exhibit binding to the host cells. To determine the effects of Acanthamoeba on the host cell cycle, a cell-cycle-specific gene array was used. This screened for 96 genes specific for host cell-cycle regulation. It was observed that Acanthamoeba inhibited expression of genes encoding cyclins F and G1 and cyclin-dependent kinase 6, which are proteins important for cell-cycle progression. Moreover, upregulation was observed of the expression of genes such as GADD45A and p130 Rb, associated with cell-cycle arrest, indicating cell-cycle inhibition. Next, the effect of Acanthamoeba on retinoblastoma protein (pRb) phosphorylation was determined. pRb is a potent inhibitor of G1-to-S cell-cycle progression; however, its function is inhibited upon phosphorylation, allowing progression into S phase. Western blotting revealed that Acanthamoeba abolished pRb phosphorylation leading to cell-cycle arrest at the G1-to-S transition. Taken together, these studies demonstrated for the first time that Acanthamoeba inhibits the host cell cycle at the transcriptional level, as well as by modulating pRb phosphorylation using host cell-signalling mechanisms. A complete understanding of Acanthamoeba-host cell interactions may help in developing novel strategies to treat Acanthamoeba infections.

  13. Occurrence of Potentially Pathogenic Bacterial-Endosymbionts in Acanthamoeba Spp.

    Directory of Open Access Journals (Sweden)

    Maryam Niyyati

    2015-06-01

    Full Text Available Acanthamoeba- bacteria interactions enable pathogenic bacteria to tolerate harsh conditions and lead to transmission to the susceptible host. The present study was aimed to address the presence of bacterial endosymbionts of Acanthamoeba isolated from recreational water sources of Tehran, Iran. To the best of our knowledge this is the first study regarding occurrence of bacteria in environmental Acanthamoeba spp. in Iran.A total of 75 samples of recreational water sources were collected. Samples were cultured on non- nutrient agar 1.5% plates. Positive Acanthamoeba spp. were axenically grown. DNA extraction and PCR reaction was performed using JDP1-2 primers. All positive samples of Acanthamoeba were examined for the presence of endosymbionts using staining and molecular methods. The PCR products were then sequenced in order to determine the genotypes of Acanthamoeba and bacteria genera.Out of 75 samples, 16 (21.3% plates were positive for Acanthamoeba according to the morphological criteria. Molecular analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes. Five isolates (35.7% were positive for bacterial endosymbionts using staining method and PCR test. Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens.The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection. To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide.

  14. Nuevos mecanismos protectores en la enfermedad inflamatoria intestinal sistema endocannabinoide y haptoglobina /

    OpenAIRE

    Márquez Mosquera, Lucía

    2012-01-01

    La Enfermedad Inflamatoria Intestinal (EII) (enfermedad de Crohn (EC) y Colitis Ulcerosa (CU)), es una enfermedad crónica, poligénica y compleja, de curso recurrente, que afecta al tracto gastrointestinal. Estudios recientes in vitro y modelos experimentales en animales sugieren que el Sistema Endocannabinoide (SE) y la Haptoglobina (Hp) pueden tener un papel protector en la EII. Estudio 1: Sistema Endocannabinoide y EII Se analizó, mediante Western-Blot e inmunohistoquímica, la presencia de ...

  15. Caracterización de la respuesta inflamatoria en la neumonía comunitaria grave. Efecto de los corticoides en la contención de dicha respuesta y en el curso evolutivo de la infección

    OpenAIRE

    Fernández Serrano, Silvia

    2012-01-01

    [spa] La neumonía adquirida en la comunidad (NAC) continúa siendo en la actualidad una enfermedad potencialmente grave, que con frecuencia presenta una evolución desfavorable, a pesar del tratamiento antibiótico adecuado. La hipótesis de la tesis plantea que es posible monitorizar la respuesta inflamatoria que tiene lugar durante la NAC y, que dicha respuesta, puede ser influenciada por los antibióticos utilizados en su tratamiento, así como modulada mediante una terapia adyuvante con co...

  16. In vitro comparative assessment of different viability assays in Acanthamoeba castellanii and Acanthamoeba polyphaga trophozoites.

    Science.gov (United States)

    Heredero-Bermejo, I; Copa-Patiño, J L; Soliveri, J; Gómez, R; de la Mata, F J; Pérez-Serrano, J

    2013-12-01

    The species of the genus Acanthamoeba are opportunistic protozoan parasites that cause different diseases in humans, such as amoebic keratitis and granulomatous encephalitis. The rise in the rate of Acanthamoeba keratitis, mainly due to the increase in contact lens wearers, turns the development of viability assays using a multi-well plate reader as a tool for screening new antiamoebic agents in vitro into an important goal. In our study, the viability assays PrestoBlue®, resazurin sodium salt, 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) and CellTiter96® were tested for their suitability as time-saving alternatives to the classical manual or direct-counting method, assessing the effect of the antiamoebic agent chlorhexidine digluconate and temperature on Acanthamoeba castellanii (ATCC® 30234™) and Acanthamoeba polyphaga 2961. Although resazurin and MTT have already been previously used in amoeba viability assays to test the activities of antiamoebic agents in vitro, it is the first time that PrestoBlue® and CellTiter96® are used for this purpose. Results indicated that the viability assays were strain-dependent leading in some cases to an overestimation of the real situation of viable cells. This implies that each viability assay ought to be set up for each amoeba strain studied.

  17. Parachlamydia acanthamoeba is endosymbiotic or lytic for Acanthamoeba polyphaga depending on the incubation temperature.

    Science.gov (United States)

    Greub, Gilbert; La Scola, Bernard; Raoult, Didier

    2003-06-01

    Parachlamydiaceae are potential emerging pathogens that naturally infect free-living amoebae. We investigated the affects of incubation temperature on the growth and cytopathic effect of P. acanthamoeba in Acanthamoeba polyphaga. A. polyphaga were infected with P. acanthamoeba and incubated at different temperatures for ten days. Bacterial growth was quantified by real-time PCR. Cytopathic effects were determined by counting the number of cysts and viable amoebae (unstained with trypan blue) in Nageotte counting chambers. Uninfected amoebae cultures were used as negative control. At 32, 35, and 37 degrees C, we observed a significant decrease in the number of viable A. polyphaga that contrasted with the delayed and smaller decrease in the number of living A. polyphaga observed at 25, 28, and 30 degrees C. Higher incubation temperature, which is associated with amoebal lysis, surprisingly was not associated with increased growth rate. P. acanthamoeba is lytic for A. polyphaga at 32-37 degrees C but endosymbiotic at 25-30 degrees C. This suggests that A. polyphaga may be a reservoir of endosymbionts at the lower temperature of the nasal mucosa, which may be liberated by lysis at higher temperature, for instance, when the amoeba is inhaled and reaches the lower respiratory tract.

  18. Pleuresia inflamatoria en la infancia: Conducta quirúrgica

    OpenAIRE

    Arturo L. Delgado Delgado; Ángel Elizechea Hernández; Lucrecia Medina Vic; Nelson Ponce Sandoval

    2003-01-01

    La neumonía complicada por pleuresía ha sido siempre un proceso que ha afectado la morbilidad y mortalidad en la edad pediátrica y en ocasiones necesita de algún proceder quirúrgico para su solución. La decisión inicial de si realizar solo una punción pleural o agregarle una pleurostomía mínima, puede ser controversial, por lo que se decidió realizar un estudio de todos aquellos niños que fueron ingresados en el Hospital Pediátrico Universitario de San Miguel del Padrón con neumonía complicad...

  19. Nuevos esteroides en enfermedad inflamatoria intestinal: uso de la budesonida.

    OpenAIRE

    José Humberto Arango; Gustavo Mariño; Abraham Kestenberg

    2009-01-01

    Este artículo revisa las aplicaciones de los esteroides en el manejo de la colitis ulcerativa, con énfasis en la budesonida. La budesonida se diseñó para disminuir los efectos adversos, gracias a su alto metabolismo de primer paso por el hígado. El uso tópico de este esteroide es una alternativa en el manejo de los pacientes con colitis ulcerativa distal.

  20. Nuevos esteroides en enfermedad inflamatoria intestinal: uso de la budesonida.

    Directory of Open Access Journals (Sweden)

    José Humberto Arango

    2009-10-01

    Full Text Available Este artículo revisa las aplicaciones de los esteroides en el manejo de la colitis ulcerativa, con énfasis en la budesonida. La budesonida se diseñó para disminuir los efectos adversos, gracias a su alto metabolismo de primer paso por el hígado. El uso tópico de este esteroide es una alternativa en el manejo de los pacientes con colitis ulcerativa distal.

  1. Sepsis por shigella flexneri

    Directory of Open Access Journals (Sweden)

    César Cabrera C

    2005-04-01

    Full Text Available Se presenta un caso raro de sepsis por Shigella flexneri en una paciente de 45 años de edad quien estando hospitalizada para el estudio de un tumor cerebral, requirió el uso de manitol y dosis altas de corticoides; luego de ello presenta deposiciones líquidas con moco y sangre, desarrolla síndrome de respuesta inflamatoria sistémica, luego se aísla Shigella flexneri en el hemocultivo; recibió tratamiento antibiótico con ciprofloxacina. Se describen las características del caso y se comenta de acuerdo con la revisión de literatura.

  2. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment

    Directory of Open Access Journals (Sweden)

    Lorenzo-Morales Jacob

    2015-01-01

    Full Text Available Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. In view of the devastating nature of this disease, here we present our current understanding of Acanthamoeba keratitis and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be potential targets for improved diagnosis, therapeutic interventions and/or for the development of preventative measures. Novel molecular approaches such as proteomics, RNAi and a consensus in the diagnostic approaches for a suspected case of Acanthamoeba keratitis are proposed and reviewed based on data which have been compiled after years of working on this amoebic organism using many different techniques and listening to many experts in this field at conferences, workshops and international meetings. Altogether, this review may serve as the milestone for developing an effective solution for the prevention, control and treatment of Acanthamoeba infections.

  3. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment.

    Science.gov (United States)

    Lorenzo-Morales, Jacob; Khan, Naveed A; Walochnik, Julia

    2015-01-01

    Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. In view of the devastating nature of this disease, here we present our current understanding of Acanthamoeba keratitis and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be potential targets for improved diagnosis, therapeutic interventions and/or for the development of preventative measures. Novel molecular approaches such as proteomics, RNAi and a consensus in the diagnostic approaches for a suspected case of Acanthamoeba keratitis are proposed and reviewed based on data which have been compiled after years of working on this amoebic organism using many different techniques and listening to many experts in this field at conferences, workshops and international meetings. Altogether, this review may serve as the milestone for developing an effective solution for the prevention, control and treatment of Acanthamoeba infections.

  4. Diversity and Seasonal Impact of Acanthamoeba Species in a Subtropical Rivershed

    Directory of Open Access Journals (Sweden)

    Po-Min Kao

    2013-01-01

    Full Text Available This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in this study, 16 (11.7% of which contained Acanthamoeba species. Samples with the highest percentage of Acanthamoeba (32.4% were obtained during the summer season, mainly from upstream areas. The identified species in the four seasons included Acanthamoeba palestinensis (T2, Acanthamoeba sp. IS2/T4 (T4, Acanthamoeba lenticulata (T5, Acanthamoeba hatchetti (T11, Acanthamoeba healyi (T12, and Acanthamoeba jacobsi (T15. The most frequently identified Acanthamoeba genotype was T4 (68.7%. Acanthamoeba genotype T4 is responsible for Acanthamoeba keratitis and should be considered for associated human health risk potential in the rivershed.

  5. Diversity and seasonal impact of Acanthamoeba species in a subtropical rivershed.

    Science.gov (United States)

    Kao, Po-Min; Chou, Ming-Yuan; Tao, Chi-Wei; Huang, Wen-Chien; Hsu, Bing-Mu; Shen, Shu-Min; Fan, Cheng-Wei; Chiu, Yi-Chou

    2013-01-01

    This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in this study, 16 (11.7%) of which contained Acanthamoeba species. Samples with the highest percentage of Acanthamoeba (32.4%) were obtained during the summer season, mainly from upstream areas. The identified species in the four seasons included Acanthamoeba palestinensis (T2), Acanthamoeba sp. IS2/T4 (T4), Acanthamoeba lenticulata (T5), Acanthamoeba hatchetti (T11), Acanthamoeba healyi (T12), and Acanthamoeba jacobsi (T15). The most frequently identified Acanthamoeba genotype was T4 (68.7%). Acanthamoeba genotype T4 is responsible for Acanthamoeba keratitis and should be considered for associated human health risk potential in the rivershed.

  6. FACTOR NUCLEAR κB (NF-κB: SIGNALOSOMA Y SU IMPORTANCIA EN ENFERMEDADES INFLAMATORIAS Y CÁNCER

    Directory of Open Access Journals (Sweden)

    Nancy P Echeverri R

    2008-06-01

    Full Text Available El factor nuclear κB (NF-κB es un dímero constituido por proteínas de la familia Rel. El NF-κB se encuentra en el citoplasma unido a proteínas inhibidoras (IkB. Las IkB son fosforiladas por diferentes cinasas que hacen parte del signalosoma como las cinasas de IKKα e IKKP y el modulador esencial de NF-κB (NEMO, la proteína cinasa activadora de mitosis (MAPK o p38 y la cinasa inductora de NF-κB (NIK. Estas cinasas al ser activadas por señales dependientes de citocinas y luz ultravioleta, fosforilan las IkB provocando su ubiquitinación, su degradación por proteosoma y la subsecuente liberación y translocación al núcleo de NF-κB. Recientemente se le ha dado una gran importancia al NF- k B en la vía de señalización desencadenada por estrés oxidativo, estrés genotóxico y daño en el DNA. A diferencia de la vía denominada clásica, en esta ruta ocurre una SUMOilación de NEMO y translocación al núcleo. En el núcleo NEMO interactúa con la proteína de la ataxia telangiectasia mutada (ATM activada en respuesta a modificaciones en la cromatina y daño en el DNA. El complejo ATM/NEMO es translocado al citoplasma donde la ATM fosforila a las IKK llevando a la ubiquitinación y posterior liberación de NF-κB que es translocado al núcleo. NF-κB desencadena procesos de supervivencia incluyendo el aumento de la transcripción de enzimas antioxidantes como la superóxido dismutasa, catalasa y glutatión. Estas enzimas participan en el control de los niveles de especies reactivas de oxígeno en la célula. La sobreactivación de NF-κB se relaciona con inflamación y cáncer. En la actualidad se desarrolla una búsqueda de fármacos que actúen sobre moléculas del signalosoma de NF-κB, no sólo para el manejo de enfermedades inflamatorias sino también para el uso durante el tratamiento de tumores resistentes a radio y quimioterapia.

  7. Gene discovery in the Acanthamoeba castellanii genome

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iain J.; Watkins, Russell F.; Samuelson, John; Spencer,David F.; Majoros, William H.; Gray, Michael W.; Loftus, Brendan J.

    2005-08-01

    Acanthamoeba castellanii is a free-living amoeba found in soil, freshwater, and marine environments and an important predator of bacteria. Acanthamoeba castellanii is also an opportunistic pathogen of clinical interest, responsible for several distinct diseases in humans. In order to provide a genomic platform for the study of this ubiquitous and important protist, we generated a sequence survey of approximately 0.5 x coverage of the genome. The data predict that A. castellanii exhibits a greater biosynthetic capacity than the free-living Dictyostelium discoideum and the parasite Entamoeba histolytica, providing an explanation for the ability of A. castellanii to inhabit adversity of environments. Alginate lyase may provide access to bacteria within biofilms by breaking down the biofilm matrix, and polyhydroxybutyrate depolymerase may facilitate utilization of the bacterial storage compound polyhydroxybutyrate as a food source. Enzymes for the synthesis and breakdown of cellulose were identified, and they likely participate in encystation and excystation as in D. discoideum. Trehalose-6-phosphate synthase is present, suggesting that trehalose plays a role in stress adaptation. Detection and response to a number of stress conditions is likely accomplished with a large set of signal transduction histidine kinases and a set of putative receptorserine/threonine kinases similar to those found in E. histolytica. Serine, cysteine and metalloproteases were identified, some of which are likely involved in pathogenicity.

  8. [Acanthamoeba spp. as opportunistic pathogens parasites].

    Science.gov (United States)

    Castrillón, Juan C; Orozco, Lina P

    2013-04-01

    Among free-living amoeba in nature, species of the genus Acanthamoeba have been associated with human disease. These amoeba are among the most abundant protozoa in nature due to its cosmopolitan distribution and are able to survive in a wide variety of habitats because its low demand for food and in harsh environments by forming structures known as cysts. However, ecological changes and incursion of its different habitats have made this organism can invade a host and live as parasites within him. That's why this type of protozoa are known as amphizoic organism, because human can be constituted as its host, causing infections in the central nervous system, disseminated infections in skin and lungs, and keratitis. Thus, since an increase in the number of cases of Acanthamoeba infections has occurred worldwide, these protozoa have become increasingly important as agents of human disease. This review summarizes what is known of this kind of free-living amoeba, focusing on the biology, ecology, pathogenesis, diagnosis, treatment and human defense mechanism against infection by the amoeba.

  9. Raman microspectroscopy analysis in the treatment of acanthamoeba keratitis.

    Directory of Open Access Journals (Sweden)

    Giulia Rusciano

    Full Text Available Acanthamoeba keratitis is a rare but serious corneal disease, often observed in contact lens wearers. Clinical treatment of infected patients frequently involves the use of polyhexamethylene biguanide (PHMB, a polymer used as a disinfectant and antiseptic, which is toxic also for the epithelial cells of the cornea. Prompt and effective diagnostic tools are hence highly desiderable for both starting early therapy and timely suspension of the treatment. In this work we use Raman microspectroscopy to analyse in vitro a single Acanthamoeba cell in cystic phase. In particular, we investigate the effect of PHMB at the single-cell level, providing useful information on both the underlying biochemical mechanism and the time frame for Acanthamoeba eradication in ocular infections. Furthermore, we demonstrate that Raman spectroscopy, in conjunction with standard multivariate analysis methods, allows discriminating between live and dead Acanthamoebas, which is fundamental to optimizing patients' treatment.

  10. Chlorhexidine monotherapy with adjunctive topical corticosteroids for acanthamoeba keratitis

    Directory of Open Access Journals (Sweden)

    Firoozeh Rahimi

    2015-01-01

    Conclusion: Chlorhexidine is effective for monotherapy in AK and could be a good choice for initiating treatment. After the initial response to anti-Acanthamoeba agents, corticosteroids can be used as adjunctive therapy depending on the clinical condition.

  11. Advances in the Diagnosis and Treatment of Acanthamoeba Keratitis

    Directory of Open Access Journals (Sweden)

    Benjamin Clarke

    2012-01-01

    Full Text Available This paper aims to review the recent literature describing Acanthamoeba keratitis and outline current thoughts on pathogenesis, diagnosis, and treatment as well as currently emerging diagnostic and treatment modalities.

  12. Acanthamoeba keratitis-A report of two cases

    Directory of Open Access Journals (Sweden)

    Girija T

    1992-01-01

    Full Text Available Two cases of unilateral corneal ulcers caused by Acanthamoeba are reported. Neither of the patients had contact lenses at any time. The diagnosis was confirmed by Giemsa stain and cultures of the corneal scrapings.

  13. Radial keratoneuritis as a presenting sign in Acanthamoeba keratitis

    Directory of Open Access Journals (Sweden)

    Abdullah Alfawaz

    2011-01-01

    Full Text Available The visual outcomes of Acanthamoeba keratitis, a rare cause of corneal infection, can be devastating. This paper reports two contact lens wearers with severe pain and photophobia who presented to the emergency room. Biomicroscopy revealed radial keratoneuritis in both individuals. Tissue culture on a nonnutrient agar plate with Escherichia coli overlay resulted in a heavy growth of Acanthamoeba. The inpatient treatment included 0.02% polyhexamethylene biguanide, chlorhexidine, neomycin/polymyxin B/bacitracin (Neosporin, and oral fluconazole, which successfully controlled the corneal infection and improvement in the best corrected visual acuity in both patients. Infection did not recur during the 12-month follow-up period. Acanthamoeba keratitis can present as radial keratoneuritis, mimicking other common corneal infections resulting in diagnostic and treatment delays. Early diagnosis and prudent treatment of Acanthamoeba keratitis are the keys to restoring vision and avoiding the subsequent need for penetrating keratoplasty.

  14. Acanthamoeba encephalitis: A Case Report and Review of Therapy

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    A Zamora

    2014-01-01

    Full Text Available Background: Acanthamoeba is a rare cause of encephalitis yet is associated with high mortality. Treatment protocols vary greatly and generally include combination therapy across a wide spectrum of antiinfective classes. Case Description: A 63-year-old male who underwent renal transplantation presented 6 months after transplantation with depressed level of consciousness. Imaging of the head with computerized tomography showed an enhancing lesion suspicious for brain abscess. Biopsy of the lesion showed Acanthamoeba cysts. The patient was treated with sulfadiazine, fluconazole, flucytosine, azithromycin, and miltefosine but without success. We review recently published cases of Acanthamoeba encephalitis with an emphasis on treatment protocols and outcomes. Conclusion: Free-living protozoans such as Acanthamoeba are ubiquitous in the environment and should be suspected in immunosuppressed persons who present with central nervous system findings and brain abscess. Biopsy is critical to establish the etiology so that appropriate combination therapy can be deployed.

  15. Current advances in diagnostic methods of Acanthamoeba keratitis

    Institute of Scientific and Technical Information of China (English)

    Wang Yuehua; Feng Xianmin; Jiang Linzhe

    2014-01-01

    Objective The objective of this article was to review the current advances in diagnostic methods for Acanthamoeba keratitis (AK).Data sources Data used in this review were retrieved from PubMed (1970-2013).The terms "Acanthamoeba keratitis" and "diagnosis" were used for the literature search.Study selection Data from published articles regarding AK and diagnosis in clinical trials were identified and reviewed.Results The diagnostic methods for the eight species implicated in AK were reviewed.Among all diagnostic procedures,corneal scraping and smear examination was an essential diagnostic method.Polymerase chain reaction was the most sensitive and accurate detection method.Culturing of Acanthamoeba was a reliable method for final diagnosis of AK.Confocal microscopy to detect Acanthamoeba was also effective,without any invasive procedure,and was helpful in the early diagnosis of AK.Conclusion Clinically,conjunction of various diagnostic methods to diagnose AK was necessary.

  16. Raman microspectroscopy analysis in the treatment of acanthamoeba keratitis.

    Science.gov (United States)

    Rusciano, Giulia; Capriglione, Paola; Pesce, Giuseppe; Del Prete, Salvatore; Cennamo, Gilda; Di Cave, David; Cerulli, Luciano; Sasso, Antonio

    2013-01-01

    Acanthamoeba keratitis is a rare but serious corneal disease, often observed in contact lens wearers. Clinical treatment of infected patients frequently involves the use of polyhexamethylene biguanide (PHMB), a polymer used as a disinfectant and antiseptic, which is toxic also for the epithelial cells of the cornea. Prompt and effective diagnostic tools are hence highly desiderable for both starting early therapy and timely suspension of the treatment. In this work we use Raman microspectroscopy to analyse in vitro a single Acanthamoeba cell in cystic phase. In particular, we investigate the effect of PHMB at the single-cell level, providing useful information on both the underlying biochemical mechanism and the time frame for Acanthamoeba eradication in ocular infections. Furthermore, we demonstrate that Raman spectroscopy, in conjunction with standard multivariate analysis methods, allows discriminating between live and dead Acanthamoebas, which is fundamental to optimizing patients' treatment.

  17. Participación del factor transcripcional NF-KB en la endocitosis de staphylococcus aureus en células de epitelio marino bovino estimuladas por prolactina

    OpenAIRE

    Lara Zárate, Leticia

    2011-01-01

    La mastitis bovina es considerada una de las enfermedades de mayor relevancia en la ganadería lechera a nivel mundial. Esta patología se caracteriza por una respuesta inflamatoria de la glándula mamaria de origen multifactorial, ocasionada principalmente por infecciones bacterianas.

  18. Several staining techniques to enhance the visibility of Acanthamoeba cysts.

    Science.gov (United States)

    El-Sayed, Nagwa Mostafa; Hikal, Wafaa Mohamed

    2015-03-01

    Acanthamoeba is one of the most common free-living amoebae. It is widespread in the environment and can infect humans causing keratitis. Delayed diagnosis or misdiagnosis leads to extensive corneal inflammation and profound visual loss. Therefore, accurate and rapid diagnosis of Acanthamoeba keratitis is essential for successful treatment and good prognosis. This study was designed to use different staining techniques to facilitate the identification of Acanthamoeba cysts. Acanthamoeba cysts were isolated by cultivation of either corneal scraping specimens or tap water samples onto non-nutrient agar plates seeded with Escherichia coli. Subcultures were done from positive cultures until unique cysts were isolated. Acanthamoeba cysts were stained temporarily using iodine, eosin, methylene blue, and calcofluor white (CFW) stains and as permanent slides after processing for mounting using modified trichrome, Gimenez and Giemsa staining. These stains were compared on the basis of staining quality including clarity of morphological details, differentiation between cytoplasm and nuclei, color and contrast, and also other characteristics of the staining techniques, including ease of handling, time taken for the procedure, and cost effectiveness. The cysts of Acanthamoeba were recognized in the form of double-walled cysts: the outer wall (ectocyst) that was being differentiated from the variably stained surrounding background and the inner wall (endocyst) that was sometimes stellated, polygonal, round, or oval and visualized as separate from the spherical, sometimes irregular, outline of the ectocyst. Regarding the temporary stains, it was found that they were efficient for visualizing the morphological details of Acanthamoeba cysts. In CFW staining, Acanthamoeba cysts appeared as bluish-white or turquoise oval halos although the internal detail was not evident. On the other hand, the results of permanent-stained slides showed the most consistent stain for identification of

  19. Ex vivo laser confocal microscopy findings of cultured Acanthamoeba trophozoites

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    Yamazaki N

    2012-08-01

    Full Text Available Natsuko Yamazaki,1 Akira Kobayashi,1 Hideaki Yokogawa,1 Yasuhisa Ishibashi,2 Yosaburo Oikawa,3 Masaharu Tokoro,4 Kazuhisa Sugiyama11Department of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan; 2Department of Ophthalmology, East Washinomiya Hospital, Kuki, Japan; 3Department of Medical Zoology, Kanazawa Medical University, Kahoku, Japan; 4Department of Parasitology, Kanazawa University Graduate School of Medical Science, Kanazawa, JapanPurpose: The purpose of the current study was to investigate ex vivo laser confocal microscopic findings of cultured Acanthamoeba trophozoites obtained from Acanthamoeba keratitis patients.Methods: Eight cultured samples of Acanthamoeba trophozoites from eight eyes of seven patients (mean age, 26.9 years; age range, 18–52 years were used. Seven samples were from corneal scrapings of Acanthamoeba keratitis patients and one sample was from the solution in a soft contact lens case. Ex vivo laser confocal microscopy was performed to qualitatively evaluate the shape and degree of light reflection of the living Acanthamoeba trophozoites.Results: Ex vivo laser confocal microscopy demonstrated highly reflective, high-contrast Acanthamoeba trophozoites with no walls (mean size, 25.4 µm; range, 17.1–58.5 µm. The shapes of the trophozoites were highly pleomorphic, and some showed characteristic acanthopodia by laser confocal microscopy.Conclusion: Ex vivo laser confocal microscopy was effective in demonstrating cultured Acanthamoeba trophozoites of various shapes and sizes. The observations of the current study may be helpful when similar structures are identified under in vivo conditions.Keywords: Acanthamoeba, trophozoite, laser confocal microscopy

  20. Crescent bodies of Parachlamydia acanthamoeba and its life cycle within Acanthamoeba polyphaga: an electron micrograph study.

    Science.gov (United States)

    Greub, Gilbert; Raoult, Didier

    2002-06-01

    Parachlamydiaceae are endosymbionts of free-living amoeba first identified in 1997. Two developmental stages, elementary and reticulate bodies, were observed; however, their localization and proportions according to culture condition and duration remain unknown. The life cycle of Parachlamydia acanthamoeba within Acanthamoeba polyphaga was studied by transmission electron microscopy of 8-, 36-, and 144-h coculture. Morphometry and quantification were performed using SAMBA software. The elementary body, the predominant stage within the amoebae, was located mainly within their vacuoles. The multiplication of Parachlamydia bacteria by binary fission of reticulate bodies was independently associated with culture in PYG broth (odds ratio [OR] = 4.4; 95% confidence interval [CI], 1.55 to 12.46) and with the presence of reticulate bodies within the amoebae (OR = 2.10; 95% CI, 1.53 to 2.89). A third developmental stage was observed, the crescent body. Its presence outside and inside the amoebae was associated mainly with prolonged incubation time (OR = 3.98; 95% CI, 1.49 to 10.68, and OR = 5.98; 95% CI, 1.75 to 20.4, respectively). Elementary and crescent bodies were released into the extracellular medium within vesicles or after amoebal lysis. For both, phagocytosis was their mode of entry. This electron micrograph study revealed another infective developmental stage, the crescent body, and provided quantitative analysis of the life cycle of P. acanthamoeba within A. polyphaga.

  1. Tratamiento medicoquirúrgico en la enfermedad inflamatoria pélvica aguda

    OpenAIRE

    Rosalba Roque González; Evelio Ramírez Hernández; Armando Leal Mursulí; Hernán Pérez Oramas; Nélida Ramos Díaz; Esteban Goitizolo Vildostegui

    2000-01-01

    Se revisaron 267 historias clínicas de pacientes ingresadas de urgencia con el diagnóstico de enfermedad inflamatoria pélvica aguda que aparece frecuentemente entre los 15 y 24 años de edad. El dolor abdominal, la leucorrea y la leucocitosis, constituyen los síntomas, signos y resultados de laboratorio más frecuentes de esta entidad. Los medios diagnósticos más utilizados fueron el ultrasonido y la laparoscopia. Los hallazgos laparoscópicos fueron confirmados en el acto quirúrgico en la mayor...

  2. Nutrición y enfermedad inflamatoria intestinal Nutrition and inflammatory bowel disease

    OpenAIRE

    2007-01-01

    En este artículo se revisan las causas y consecuencias de los déficits nutricionales en la enfermedad inflamatoria intestinal, con especial énfasis en sus implicaciones fisiopatológicas. Además, se sientan las bases de la intervención nutricional en estos pacientes tanto desde la perspectiva del consejo dietético como del uso de soporte nutricional especializado. Se hace un especial hincapié en la discusión de las evidencias existentes en favor de un papel terapéutico primario de la nutrición...

  3. Statins and voriconazole induce programmed cell death in Acanthamoeba castellanii.

    Science.gov (United States)

    Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Sifaoui, Ines; Reyes-Batlle, María; Valladares, Basilio; Martínez-Carretero, Enrique; Piñero, José E; Maciver, Sutherland K; Lorenzo-Morales, Jacob

    2015-05-01

    Members of the genus Acanthamoeba are facultative pathogens of humans, causing a sight-threatening keratitis and a life-threatening encephalitis. In order to treat those infections properly, it is necessary to target the treatment not only to the trophozoite but also to the cyst. Furthermore, it may be advantageous to avoid parasite killing by necrosis, which may induce local inflammation. We must also avoid toxicity of host tissue. Many drugs which target eukaryotes are known to induce programmed cell death (PCD), but this process is poorly characterized in Acanthamoeba. Here, we study the processes of programmed cell death in Acanthamoeba, induced by several drugs, such as statins and voriconazole. We tested atorvastatin, fluvastatin, simvastatin, and voriconazole at the 50% inhibitory concentrations (IC50s) and IC90s that we have previously established. In order to evaluate this phenomenon, we investigated the DNA fragmentation, one of the main characteristics of PCD, with quantitative and qualitative techniques. Also, the changes related to phosphatidylserine exposure on the external cell membrane and cell permeability were studied. Finally, because caspases are key to PCD pathways, caspase activity was evaluated in Acanthamoeba. All the drugs assayed in this study induced PCD in Acanthamoeba. To the best of our knowledge, this is the first study where PCD induced by drugs is described quantitatively and qualitatively in Acanthamoeba.

  4. Isolation of Acanthamoeba spp. from Different Environmental Sources

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    A Motevalli Haghi

    2008-04-01

    Full Text Available Background: Acanthamoeba spp. are free-living amebas found in a wide variety of natural habitats. The high percentage of Acanthamoeba in different environmental sources represents a sanitary risk for public health especially contact lens users and immunocompromised patients. The aim of this study was to determine the presence of Acanthamoeba spp. in different environments such as water, soil, dust and ophthalmology wards. Methods: From March to November 2007, 80 samples were collected from numerous localities in Tehran city including university campus, Laleh park and ophthalmology center. Sample types were water, soil, dust, cow faeces and medical in­strument. Each sample was filtered through nitrate membrane and cultured on 1% non-nutrient agar. These plates were fol­lowed up daily for 2 weeks. Monitoring continued for two months on a weekly basis.  Results: Overall, 46.25% of samples contained Acanthamoeba spp. All of the soil samples had shown positive culture in contrast to tap water. Of 61 dust samples, 28 were positive. Interestingly, we were able to isolate Acanthamoeba in treat­ment unit of an ophthalmology center in Tehran. It should be mentioned that two cow faeces showed positive culture as well.Conclusion: The widespread distribution of Acanthamoeba spp. across the environmental sources and increasing numbers of HIV+ patients and contact lens wearers, as well as its ability as a pathogen carrier for humans, demands  more awareness and knowledge for public as a risk for human health.

  5. Genotyping of clinical isolates of Acanthamoeba genus in Venezuela.

    Science.gov (United States)

    Wagner, Carolina; Reyes-Batlle, María; Ysea, María Alejandra Vethencourt; Pérez, Mónica V Galindo; de Rondón, Carmen Guzmán; Paduani, Anaibeth J Nessi; Pérez, Angelyseb Dorta; López-Arencibia, Atteneri; Sifaoui, Ines; de Galindo, María Virginia Pérez; de Suárez, Eva Pérez; Martínez-Carretero, Enrique; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2016-12-01

    Free-living amoebae of Acanthamoeba genus are opportunistic pathogens distributed worldwide. Strains included in this genus are causative agents of a fatal encephalitis and a sight-threating keratitis in humans and other animals. In this study, 550 clinical samples which were collected between 1984 and 2014 from different patients with suspected infections due to Acanthamoeba were initially screened for the presence of this amoebic genus at the Laboratorio de Amibiasis-Escuela de Bioanálisis at the Universidad Central de Venezuela. Samples were cultured in 2% Non-Nutrient agar plates seeded with a layer of heat killed Escherichia coli. From the 550 clinical samples included in this study, 18 of them were positive for Acanthamoeba genus after culture identification. Moreover, positive samples were confirmed after amplification of the Diagnostic Fragment 3 (DF3) of the Acanthamoeba18S rDNA genus and sequencing was carried out in order to genotype the isolated strains of Acanthamoeba. Furthermore, the pathogenic potential of the strains was checked by performing thermotolerance and osmotolerance assays. Sequencing of the DF3 region resulted in the identification of genotype T4 in all the isolated strains. Moreover, most isolates were thermotolerant or both thermotolerant and osmotolerant and thus were classified as potentially pathogenic strains. To the best of our knowledge, this is the first report on the molecular characterization at the genotype level of Acanthamoeba strains in Venezuela.

  6. Isolation of Acanthamoeba from the rhizosphere of maize and lucerne plants

    Science.gov (United States)

    Orosz, Erika; Farkas, Ágnes; Ködöböcz, László; Becsak, Péter; Danka, József; Kucsera, István; Füleky, György

    2013-04-01

    Acanthamoeba species are free-living amoebae that can be found in almost every range of environments. Within this genus, a number of species are recognized as human pathogens, potentially causing Acanthamoeba keratitis, granulomatous amoebic encephalitis, and chronic granulomatous lesions. Soil and water samples were taken from experimental station at Julianna Major of Plant Protection Institute of Centre for Agricultural Research, Hungarian Academy of Sciences. We detected living Acanthamoeba spp. based on culture- confirmed detection combined with the molecular taxonomic identification method. Living Acanthamoeba spp. were detected in thirteen (65%) samples. The presence of Acanthamoeba spp. in the samples depends significantly on the rhizosphere plants. The most frequently identified living Acanthamoeba genotype was T4 followed by T11, T2/T6 and T17. Genotypes T4 and T11 of Acanthamoeba, are responsible for Acanthamoeba keratitis as well as granulomatous amoebic encephalitis, and should therefore be considered as a potential health risk associated with human activities in the environment.

  7. Effects of Grazing by the Free-Living Soil Amoebae Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis on Various Bacteria

    OpenAIRE

    Weekers, Peter H. H.; Paul l.E. Bodelier; Wijen, John P. H.; Vogels, Godfried D.

    1993-01-01

    Cultures of 10 different bacteria were used to serve as food sources for axenically grown Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. The nonpigmented enterobacteriaceae Escherichia coli K-12 and Klebsiella aerogenes appeared to be excellent feed to all three amoebae. Hardly any growth or ammonium production was observed in tests with Chromatium vinosum and Serratia marcescens, which share the presence of pigmented compounds. Distinct differences in net amm...

  8. Enfermedad inflamatoria pélvica: un reto en el diagnóstico y tratamiento precoz

    OpenAIRE

    Baquedano Mainar,Laura; Lamarca Ballestero,Marta; Puig Ferrer,Fernando; Ruiz Conde,Miguel Angel

    2014-01-01

    Objetivo: Actualizar los datos disponibles en referencia a la enfermedad inflamatoria pélvica para poder unificar criterios diagnósticos y terapéuticos y así minimizar las complicaciones que a corto y/o largo plazo puedan derivar. Método: Revisión de la literatura en Pubmed atendiendo sobre todo a las guías clínicas más actualizadas y ensayos clínicos aleatorizados. Resultados: La enfermedad inflamatoria pélvica es un cuadro infeccioso común entre las mujeres en edad fértil. Su mecanismo de t...

  9. CORIOAMNIONITIS HISTOLÓGICA Y MORBIMORTALIDAD NEONATAL: APROXIMACIÓN AL SÍNDROME DE RESPUESTA INFLAMATORIA FETAL

    OpenAIRE

    Rincón Ricote,M. Inmaculada; Magdaleno Dans, Fernando; Sancha Naranjo,Marta; Omeñaca Teres,Félix; González González,Antonio

    2010-01-01

    Antecedentes: El síndrome de respuesta inflamatoria fetal (SRIF) es una entidad relacionada con la presencia de inflamación intrauterina y suele asociarse a infección intraamniótica. Su consecuencia más grave es la lesión cerebral y posterior desarrollo de parálisis cerebral. Objetivo: Evaluar la relación entre el síndrome de respuesta inflamatoria fetal y el desarrollo de complicaciones neonatales. Método: Estudio descriptivo y retrospectivo, realizado en el Hospital Universitario La Paz de ...

  10. Enfermedad inflamatoria pélvica: un reto en el diagnóstico y tratamiento precoz

    OpenAIRE

    Baquedano Mainar,Laura; Lamarca Ballestero,Marta; Puig Ferrer,Fernando; Ruiz Conde,Miguel Angel

    2014-01-01

    Objetivo: Actualizar los datos disponibles en referencia a la enfermedad inflamatoria pélvica para poder unificar criterios diagnósticos y terapéuticos y así minimizar las complicaciones que a corto y/o largo plazo puedan derivar. Método: Revisión de la literatura en Pubmed atendiendo sobre todo a las guías clínicas más actualizadas y ensayos clínicos aleatorizados. Resultados: La enfermedad inflamatoria pélvica es un cuadro infeccioso común entre las mujeres en edad fértil. Su mecanismo de t...

  11. The pathogenesis of Acanthamoeba infections: current status\\ud and future implications

    OpenAIRE

    Khan, Naveed Ahmed

    2004-01-01

    Acanthamoeba are opportunistic protozoan parasites that can cause painful, visionthreatening\\ud keratitis. However the pathogenesis and pathophysiology of\\ud Acanthamoeba keratitis remain incompletely understood. Most cases of\\ud Acanthamoeba keratitis develop as a result of poor hygiene in contact lens care but it\\ud is unclear how amoebae transmigrate from the environment into the cornea leading to\\ud inflammation, photophobia and blindness. Acanthamoeba keratitis has become\\ud increasingly...

  12. Development of Colorimetric Microtiter Plate Assay for Assessment of Antimicrobials against Acanthamoeba

    OpenAIRE

    McBride, James; Ingram, Paul R.; Henriquez, Fiona L.; Roberts, Craig W.

    2005-01-01

    We have developed and optimized a 96-well microtiter plate assay, based on the reduction of alamarBlue, to assess the efficacies of much needed new antimicrobials against Acanthamoeba species. This assay has been optimized for determination of drug efficacy against two potentially pathogenic species, Acanthamoeba castellanii and Acanthamoeba polyphaga, and has been validated by comparison of their relative susceptibilities to chlorhexidine, a drug widely used to treat Acanthamoeba keratitis. ...

  13. Encystment in Acanthamoeba castellanii: a review.

    Science.gov (United States)

    Lloyd, David

    2014-11-01

    Differentiation of Acanthamoeba castellanii trophozoites involves massive turnover of cellular components and remodelling of organelle structure and function so as to produce a cryptobiotic cell, resistant to desiccation, heat, freezing, and chemical treatments. This review presents a summary of a decade of research on the most studied aspects of the biochemistry of this process, with emphasis on problems of biocide and drug resistances, putative new targets, molecular and cell biology of the process of encystment, and the characteristics of the encysted state. As well as the intrinsic pathogenicity of the organism towards the cornea, and the ability of related species to invade the human brain, its propensity for harbouring and transmitting pathogenic bacteria and viruses is considerable and leads to increasing concerns. The long-term survival and resistance of cysts to drugs and biocides adds another layer of complexity to the problem of their elimination.

  14. Acanthamoeba castellanii cysts: new ultrastructural findings.

    Science.gov (United States)

    Chávez-Munguía, Bibiana; Salazar-Villatoro, Lizbeth; Lagunes-Guillén, Anel; Omaña-Molina, Maritza; Espinosa-Cantellano, Martha; Martínez-Palomo, Adolfo

    2013-03-01

    During Acanthamoeba castellanii trophozoite-cysts differentiation, four morphological stages were identified by scanning electron microscopy: trophozoite, precyst, immature cysts, and mature cysts. Fluorescence microscopy reveals the presence of small cumulus of actin in the cytoplasm of precysts after treatment with rhodamine phalloidin. By the contrary, in mature cysts, fluorescence was not observed. However, when excystation was induced, large fluorescent patches were present. By transmission electron microscopy, encysting amebas showed small cytoplasmic vesicles containing fibrillar material, surrounded by a narrow area of thin fibrils. Similar appearance was observed in pseudopods and phagocytic invaginations. In addition, large aggregates of rod-shape elements, similar to the chromatoid bodies, described in other amebas, were present in the cytoplasm. These cysts presented large areas with orange fluorescence after treatment with acridine orange.

  15. Ecto-ATPases of clinical and non-clinical isolates of Acanthamoeba.

    Science.gov (United States)

    Sissons, James; Alsam, Selwa; Jayasekera, Samantha; Khan, Naveed Ahmed

    2004-11-01

    Acanthamoeba are opportunistic protozoan parasites that can cause fatal granulomatous amoebic encephalitis and eye keratitis, however the pathogenic mechanisms of Acanthamoeba remain unclear. In this study, we described the ability of live Acanthamoeba to hydrolyse extracellular ATP. Both clinical and non-clinical isolates belonging to genotypes, T1, T2, T3, T4 and T7 exhibited ecto-ATPase activities in vitro. Using non-denaturing polyacrylamide gel electrophoresis, ecto-ATPases were further characterized. All Acanthamoeba isolates tested, exhibited a single ecto-ATPase band (approximate molecular weight of 272 kDa). However, clinical isolates exhibited additional bands suggesting that ecto-ATPases may play a role in the pathogenesis of Acanthamoeba. This was supported using suramin (ecto-ATPase inhibitor), which inhibited Acanthamoeba-induced host cell cytotoxicity. Previously, we and others have shown that Acanthamoeba binds to host cells using their mannose-binding protein and binding can be blocked using exogenous alpha-mannose. In this study, we observed that alpha-mannose significantly increased ecto-ATPase activities of pathogenic Acanthamoeba belonging to T1, T2, T3 and T4 genotypes but had no effect on non-pathogenic Acanthamoeba (belonging to T7 genotype). Overall, we have shown, for the first time, that Acanthamoeba exhibit ecto-ATPase activities, which may play a role in the pathogenesis of Acanthamoeba as well as their potential role in the differentiation of pathogenic Acanthamoeba.

  16. Citocinas anti-inflamatorias y sus acciones y efectos en la sepsis y el choque séptico (Anti-inflammatory cytokines and their actions and effects in the sepsis and septic shock.

    Directory of Open Access Journals (Sweden)

    González Álvarez, Ricardo

    2009-08-01

    Full Text Available ResumenLas citocinas anti- inflamatorias reconocidas son las interleucinas (IL IL-4, IL-10, IL-6, IL-13, el factor estimulante de colonia granulocito –macrófago (FEC-GM y el interferón alfa (IFN-alfa, los cuales tienen la capacidad de inhibir la liberación de las citocinas pro-inflamatorias y de inducir la producción del antagonista del receptor de la IL-1 y la liberación del receptor soluble del FNT, los cuales limitan algunas de las actividades de las citocinas pro-inflamatorias IL-1 y FNT-α. Sin embargo, los eventos que ocurren durante la inflamación y la sepsis no son tan simples como parajustificar siempre acciones antagonistas entre las citocinas proinflamatorias y las anti-inflamatorias, las cuales, pueden tener acorde con las circunstancias un comportamiento dual y/o antagónico en el choque séptico. Así por ejemplo la IL-10 tiene propiedades anti-inflamatorias, lo cual ocurre en la diabetes auto-inmune cuyo comienzo y desarrollo son acelerados en ratones transgénicos que sobre-expresan la IL-10 en los islotes pancreáticos. Esto también ocurre con la IL-10 en un modelo de uveítis en el que se incrementó la inflamación ocular. Ejemplos como los anteriores son extensivos a otras citocinas anti-inflamatorias como la IL-4, IL-13, el factor estimulante de colonia granulocito –macrófago y el factor beta transformante del crecimiento (FβTC, y que serán mencionadas y descritas en el presente artículo de revisión, así como los factores que determinan ese comportamiento dual de las citocinas.Summaryinterleucinas. The anti-inflammatory cytokines which are recognized are the interleukins (IL IL-4, IL-10, IL-13, interferon-alpha (IFN-α and because they are able to inhibit the release of pro inflammatory cytokines, to induce the production of IL-1 receptor antagonist (IL-ra and the release of soluble TNF receptor and to limit some of the pro-inflammatory activities of IL-1 and TNF. However the events which occur during

  17. Quinazolinas S-sustituidas y sus aplicaciones terapéuticas para el tratamiento de enfermedades mediadas por PDE7

    OpenAIRE

    Gil Ayuso-Gontán, Carmen; Martínez, Ana; García Fernández, Ana María; Pérez Fernández, Daniel Ignacio

    2015-01-01

    La presente invención se refiere a una familia de derivados de quinazolinas S-sustituidas que presentan actividad inhibitoria de la enzima fosfodiesterasa 7 (PDE7), útiles para el tratamiento o prevención de enfermedades mediadas por dicha enzima, especialmente enfermedades inflamatorias, neurodegenerativas, neurológicas, psiquiátricas y/o autoinmunes.

  18. Acanthamoeba spp. as agents of disease in humans.

    Science.gov (United States)

    Marciano-Cabral, Francine; Cabral, Guy

    2003-04-01

    Acanthamoeba spp. are free-living amebae that inhabit a variety of air, soil, and water environments. However, these amebae can also act as opportunistic as well as nonopportunistic pathogens. They are the causative agents of granulomatous amebic encephalitis and amebic keratitis and have been associated with cutaneous lesions and sinusitis. Immuno compromised individuals, including AIDS patients, are particularly susceptible to infections with Acanthamoeba. The immune defense mechanisms that operate against Acanthamoeba have not been well characterized, but it has been proposed that both innate and acquired immunity play a role. The ameba's life cycle includes an active feeding trophozoite stage and a dormant cyst stage. Trophozoites feed on bacteria, yeast, and algae. However, both trophozoites and cysts can retain viable bacteria and may serve as reservoirs for bacteria with human pathogenic potential. Diagnosis of infection includes direct microscopy of wet mounts of cerebrospinal fluid or stained smears of cerebrospinal fluid sediment, light or electron microscopy of tissues, in vitro cultivation of Acanthamoeba, and histological assessment of frozen or paraffin-embedded sections of brain or cutaneous lesion biopsy material. Immunocytochemistry, chemifluorescent dye staining, PCR, and analysis of DNA sequence variation also have been employed for laboratory diagnosis. Treatment of Acanthamoeba infections has met with mixed results. However, chlorhexidine gluconate, alone or in combination with propamidene isethionate, is effective in some patients. Furthermore, effective treatment is complicated since patients may present with underlying disease and Acanthamoeba infection may not be recognized. Since an increase in the number of cases of Acanthamoeba infections has occurred worldwide, these protozoa have become increasingly important as agents of human disease.

  19. A case of radial keratoneuritis in non-Acanthamoeba keratitis

    Directory of Open Access Journals (Sweden)

    Mutoh T

    2012-09-01

    Full Text Available Tetsuya Mutoh, Yukihiro Matsumoto, Makoto ChikudaDepartment of Ophthalmology, Dokkyo Medical University Koshigaya Hospital, Saitama, JapanAbstract: A case of non-Acanthamoeba keratitis with radial keratoneuritis, which is thought to be pathognomonic for Acanthamoeba keratitis, is reported. A healthy 32-year-old woman with a history of frequent replacement of her contact lenses due to wear was examined at Dokkyo Medical University Koshigaya Hospital (Saitama, Japan and found to have a slight corneal opacity that was accompanied by radial keratoneuritis. Based on both the patient’s clinical findings and past history, the presence of Acanthamoeba keratitis was highly suspected. However, direct light microscopy of corneal scrapings stained by the Parker ink–potassium hydroxide method only found Acanthamoeba-type material in the specimen collected at her initial visit. In all other specimens collected from the patient, no Acanthamoeba was found either when using the same method or when performing cultures of the surgical debridement of the corneal lesion. In addition, topical antifungal eye drops, systemic antifungal drugs, and surgical debridement were also not effective in this case. Since a precise diagnosis could not be made, the patient was treated with topical 0.1% betamethasone sodium, which ultimately resulted in a dramatic improvement of her corneal inflammation. At 23 days after initiation of topical administration of 0.1% betamethasone sodium, visual acuity was 20/250, with a slight corneal opacity noted at the original site of infection. The outcome of the current case suggests that radial keratoneuritis is not always pathognomonic for Acanthamoeba keratitis.Keywords: radial keratoneuritis, non-Acanthamoeba keratitis, topical corticosteroid

  20. Acil coenzima-a sintetasas de cadena larga (acsls) como biomarcadores citoserológicos en enfermedades inflamatorias o autoinmunes

    OpenAIRE

    Matesanz, F.; Fedetz, María; Ndagire, D.; Alcina, Antonio; Sabio, José Mario

    2010-01-01

    [ES] Uso de las Acil-Coenzima-A sintetasas de cadena larga (ACSL) como biomarcadores cito-serológicos de enfennedades inflamatorias o autoinmunes, y especialmente de lupus eritematoso sistémico, y método de obtención de datos útiles para el diagnóstico y seguimiento de dichas enfennedades.

  1. Citocinas anti-inflamatorias y sus acciones y efectos en la sepsis y el choque séptico

    National Research Council Canada - National Science Library

    Ricardo González Álvarez; Zullyt Zamora Rodríguez; Yaima Alonso González

    2009-01-01

    ResumenLas citocinas anti- inflamatorias reconocidas son las interleucinas (IL) IL-4, IL-10, IL-6, IL-13, el factor estimulante de colonia granulocito –macrófago (FEC-GM) y el interferón alfa (IFN-alfa...

  2. Cytotoxic activity of N-chlorotaurine on Acanthamoeba spp.

    Science.gov (United States)

    Fürnkranz, Ursula; Nagl, Markus; Gottardi, Waldemar; Köhsler, Martina; Aspöck, Horst; Walochnik, Julia

    2008-02-01

    Acanthamoeba spp. are the causative agents of Acanthamoeba keratitis (AK), which mainly occurs in contact lens wearers, and of skin lesions, granulomatous amoebic encephalitis (GAE), and disseminating diseases in the immunocompromised host. AK therapy is complex and irritating for the eye, skin lesions are difficult to treat, and there is no effective treatment for GAE. Therefore, new anti-Acanthamoeba drugs are needed. We investigated the anti-Acanthamoeba activity of N-chlorotaurine (NCT), an endogenous mild antiseptic. It was shown that NCT has amoebicidal qualities, both in phosphate-buffered saline (PBS) and in amoebic culture medium. After 6 h of treatment with 10 mM NCT in PBS, the levels of trophozoites of all strains investigated already showed at least a 2-log reduction. When the trophozoites were treated with 20 mM NCT in culture medium, they showed a 2-log reduction after 24 h. The addition of NH(4)Cl to NCT led to a faster decrease in the numbers of living cells, if tests were carried out in PBS. A delay of excystation was observed when cysts were treated with 55 mM (1%) NCT in culture medium. A complete failure of excystment was the result of treatment with 1% NCT plus 1% NH(4)Cl in PBS. Altogether, NCT clearly demonstrated amoebicidal activity at concentrations well tolerated by human tissues and might be useful as a topical drug for the treatment of Acanthamoeba infections. The addition of ammonium chloride can be considered to enhance the activity.

  3. Recent advances in the treatment of Acanthamoeba keratitis.

    Science.gov (United States)

    Kumar, Raman; Lloyd, David

    2002-08-15

    Infection of the eye caused by Acanthamoeba species constitutes a burgeoning and unsolved problem. Of individuals with Acanthamoeba keratitis, 85% wear contact lenses; abrasion of the cornea is implicated. Corneal infection often can be prevented by good lens care and hygiene. Severe Acanthamoeba keratitis often can be very difficult to treat; surgery can be less than successful and may lead to further problems. The encysted stage in the life cycle of Acanthamoeba species appears to cause the most problems; many biocides are ineffective in killing the highly resistant cysts. Combination therapy--that is, use of 2 or 3 biocides, sometimes with antibacterial antibiotics--appears to work best. Recurrence is common if treatment is stopped prematurely. Immunologic methods are being investigated as a form of prevention, and oral immunization of animals recently has been successful in the prevention of Acanthamoeba keratitis by inducing immunity before infection occurs. Immunization thus may eventually become the best approach for reduction of the incidence of amebic infection in humans.

  4. Expressed sequence tags (ESTs) analysis of Acanthamoeba healyi

    Science.gov (United States)

    Kong, Hyun-Hee; Hwang, Mee-Yeul; Kim, Hyo-Kyung

    2001-01-01

    Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface proteins, enzymes for DNA, energy production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba. PMID:11441502

  5. [Treatment of Acanthamoeba keratitis: possibilities, problems, and new approaches].

    Science.gov (United States)

    Walochnik, Julia; Duchêne, Michael; Eibl, Hansjörg; Aspöck, Horst

    2003-01-01

    Acanthamoeba keratitis is a corneal disease associated predominantly with contact lens wear. The occurrence of Acanthamoeba keratitis has been rising since 1990 in correlation to the growing number of contact lens wearers. To date approximately 2000 cases have been published around the world. Due to the complicated diagnostics, the elaborate treatment and the usually bad compliance of the patients, Acanthamoeba keratitis unfortunately very often takes a serious progression, which may lead to serious visual loss and perforating keratoplasty. Today, local treatment with a combination of polyhexamethylene biguanide (PHMB) and propamidine isethionate (Brolene) is considered the first line therapy for Acanthamoeba keratitis. Alternatively also a combination of propamidine and chlorhexidine or neomycine achieves good therapeutic results. However, the complicated mode of application consistently remains a problem. The intensive local treatment, i.e. hourly application of therapeutics during the first three days day and night makes hospitalization inevitable. Moreover, sufficient efficacy can not always be achieved, and also resistance against propamidine has already been observed. Recently propamidine has sometimes been replaced by hexamidine, which seems to have a greater cysticidal activity. A new path might be struck by the application of alkylphosphocholines. These are phosphocholines esterified to aliphatic alcohols. They exhibit in vitro and in vivo antineoplastic activity and have been shown to be cytotoxic against Leishmania donovani, Trypanosoma cruzi, and Entamoeba histolytica. A recent study has demonstrated that particularly hexadecylphosphocholine is highly effective also against various strains of Acanthamoeba.

  6. Protein profiles and immunoreactivities of Acanthamoeba morphological groups and genotypes.

    Science.gov (United States)

    Pumidonming, Wilawan; Koehsler, Martina; Leitsch, David; Walochnik, Julia

    2014-11-01

    Acanthamoeba is a free-living protozoan found in a wide variety of habitats. A classification of Acanthamoeba into currently eighteen genotypes (T1-T18) has been established, however, data on differences between genotypes on the protein level are scarce. The aim of this study was to compare protein and immunoreactivity profiles of Acanthamoeba genotypes. Thirteen strains, both clinical and non-clinical, from genotypes T4, T5, T6, T7, T9, T11 and T12, representing three morphological groups, were investigated for their protein profiles and IgG, IgM and IgA immunoreactivities. It was shown that protein and immunoreactivity profiles of Acanthamoeba genotypes T4, T5, T6, T7, T9, T11 and T12 are clearly distinct from each other, but the banding patterns correlate to the morphological groups. Normal human sera revealed anti-Acanthamoeba antibodies against isolates of all investigated genotypes, interestingly, however only very weak IgM and virtually no IgA immunoreactivity with T7 and T9, both representing morphological group I. The strongest IgG, IgM and IgA immunoreactivities were observed for genotypes T4, T5 and T6. Differences of both, protein and immunological patterns, between cytopathic and non-cytopathic strains, particularly within genotype T4, were not at the level of banding patterns, but rather in expression levels.

  7. Acanthamoeba keratitis: an emerging disease gathering importance worldwide?

    Science.gov (United States)

    Lorenzo-Morales, Jacob; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Arnalich-Montiel, Francisco; Piñero, José E; Valladares, Basilio

    2013-04-01

    Acanthamoeba keratitis (AK) is increasingly being recognized as a severe sight-threatening ocular infection worldwide. Although contact lens wear is the leading risk factor for AK, Acanthamoeba parasites are also an important cause of keratitis in non-contact lens wearers. Diagnosis of AK is challenging, and the available treatments are lengthy and not fully effective against all strains. The pathogenesis of Acanthamoeba is still under study, and the identification of the key factors involved in this process should be useful for the development of fully effective therapies. This review focuses on recent developments on AK pathogenesis and diagnosis as well as novel strategies for the evaluation of anti-amoebic agents that could be applied in the near future against these pathogens.

  8. Enucleation following treatment with intravenous pentamidine for Acanthamoeba sclerokeratitis

    Directory of Open Access Journals (Sweden)

    Rebecca A Kuennen

    2010-09-01

    Full Text Available Rebecca A Kuennen, Reynell Harder Smith, Thomas F Mauger, Elson CraigDepartment of Ophthalmology, The Ohio State University, Columbus, Ohio, USAPurpose: To describe the course and outcome of treatment of advanced Acanthamoeba sclerokeratitis with intravenous pentamidine.Methods: A case of advanced Acanthamoeba sclerokeratitis was resistant to conventional therapy and was treated with intravenous pentamidine. The eye was later removed due to incapacitating pain.Results: The eye showed Acanthamoeba organisms within the cornea and evidence of acute and chronic inflammation throughout the remainder of the eye. The patient has survived without orbital recurrence for 2 years.Conclusions: This case demonstrates late inflammation with active Acanthameoba keratitis following systemic pentamidine therapy.Keywords: keratitis, scleritis

  9. Acanthamoeba T4 genotype associated with keratitis infections in Tunisia.

    Science.gov (United States)

    Dendana, F; Sellami, H; Trabelsi, H; Neji, S; Cheikhrouhou, F; Makni, F; Ayadi, A

    2013-01-01

    Acanthamoeba keratitis (AK) is a sight-threatening infection. We report five cases of AK diagnosed from 2005 to 2009 in the Laboratory of Parasitology-Mycology at Habib Bourguiba Sfax Hospital, Tunisia. All were associated with improper care of contact lenses (rinsing of contact lenses with tap water and inappropriate cleaning) and lens storage. The patients displayed different clinical presentations: corneal inflammation, corneal ulceration, and corneal abscess. The diagnosis was made after direct examination, culture, and polymerase chain reaction amplification with specific primers. The genotype classification was based on the highly variable DF3 region in the 18S rRNA gene. This is the first study characterizing Acanthamoeba genotype in Tunisia and North Africa. All Acanthamoeba isolates were associated to the T4 genotype. Three different DF3 sequence types were related to AK infections T4/10, T4/15, and T4/16.

  10. ERRORES FRECUENTES EN EL DIAGNÓSTICO Y TRATAMIENTO DE LOS PACIENTES CON ENFERMEDAD INFLAMATORIA INTESTINAL

    Directory of Open Access Journals (Sweden)

    JAIME LUBASCHER C., DR.

    2015-09-01

    Full Text Available El creciente aumento de pacientes con enfermedad inflamatoria intestinal (EII, sumado a la multiplicidad de alternativas terapéuticas y a la poca experiencia en general de los médicos para tratar estas patologías, ya que son enfermedades emergentes, ha facilitado que se cometan errores tanto en el diagnóstico como en el tratamiento de las EII. En este artículo, se presentarán los más frecuentes e importantes, según la experiencia del autor, con el objeto de corregir estas conductas y alertar a los equipos médicos sobre éstas. El listado incluye errores cometidos en la historia clínica, laboratorio general, endoscopia y tratamiento.

  11. Pathogenic assays of acanthamoeba belonging to the t4 genotype.

    Directory of Open Access Journals (Sweden)

    Hamed Mirjalali

    2013-12-01

    Full Text Available Acanthamoeba genus is introduced as opportunistic and cosmopolitan parasite. Monkey and wistar rat are appropriate models for experimental study on Acanthamoeba infection. In this study Acanthamoeba spp. were isolated from hot spring (HS, windows dust (WD and a corneal sample of keratitis patient (KP and their pathogenicity surveyed by in vitro and in vivo tests.Isolates of Acanthamoeba were cultivated axenically for 12 months in PYG medium. Overall, 30 wistar rats, in 6 equal groups were used for developing experimental Acanthamoeba keratitis (AK and Granulomatous Amoebic Encephalitis (GAE. The Keratitis and Granulomatous Encephalitis experiments were performed by intrastromal and intranasal inoculation of Acanthamoeba cysts, respectively. Pathogenicity of the three isolates was also evaluated by in vitro test using osmotolerance and temperature tolerance assays. Identification of genotypes were performed by PCR technique and sequencing.None of the isolates could perform AK and GAE in wistar rats, although all isolates were described as T4 genotype. Isolates obtained from KP and WD could grow only in 30 °C, but not in 37 °C and 40 °C. On the other hand, HS isolate grew in 30 °C and 37 °C but not in 40 °C. Moreover, all of isolate grew in 0.5 M mannitol but not in 1 M and 1.5 M.T4 isolates with a long-term axenic culture and different factors related to host and parasite may play role in pathogenicity of these free-living amoebae.

  12. IL-17A-mediated protection against Acanthamoeba keratitis.

    Science.gov (United States)

    Suryawanshi, Amol; Cao, Zhiyi; Sampson, James F; Panjwani, Noorjahan

    2015-01-15

    Acanthamoeba keratitis (AK) is a very painful and vision-impairing infection of the cornea that is difficult to treat. Although past studies have indicated a critical role of neutrophils and macrophages in AK, the relative contribution of the proinflammatory cytokine, IL-17A, that is essential for migration, activation, and function of these cells into the cornea is poorly defined. Moreover, the role of the adaptive immune response, particularly the contribution of CD4(+) T cell subsets, Th17 and regulatory T cells , in AK is yet to be understood. In this report, using a mouse corneal intrastromal injection-induced AK model, we show that Acanthamoeba infection induces a strong CD4(+) T effector and regulatory T cell response in the cornea and local draining lymph nodes. We also demonstrate that corneal Acanthamoeba infection induces IL-17A expression and that IL-17A is critical for host protection against severe AK pathology. Accordingly, IL-17A neutralization in Acanthamoeba-infected wild-type mice or Acanthamoeba infection of mice lacking IL-17A resulted in a significantly increased corneal AK pathology, increased migration of inflammatory cells at the site of inflammation, and a significant increase in the effector CD4(+) T cell response in draining lymph nodes. Thus, in sharp contrast with other corneal infections such as herpes and Pseudomonas aeruginosa keratitis where IL-17A exacerbates corneal pathology and inflammation, the findings presented in this article suggest that IL-17A production after Acanthamoeba infection plays an important role in host protection against invading parasites.

  13. The role of proteases in the differentiation of Acanthamoeba castellanii.

    Science.gov (United States)

    Dudley, Ricky; Alsam, Selwa; Khan, Naveed Ahmed

    2008-09-01

    Proteases are significant determinants of protozoan pathogenicity and cytolysis of host cells. However, there is now growing evidence of their involvement in cellular differentiation. Acanthamoeba castellanii of the T4 genotype elaborates a number of proteases, which are inhibited by the serine protease inhibitor phenylmethylsulphonyl fluoride. Using this and other selective protease inhibitors, in tandem with siRNA primers, specific to the catalytic site of Acanthamoeba serine proteases, we demonstrate that serine protease activity is crucial for the differentiation of A. castellanii. Furthermore, both encystment and excystment of A. castellanii was found to be dependent on serine protease function.

  14. Acanthamoeba DNA can be directly amplified from corneal scrapings.

    Science.gov (United States)

    El-Sayed, Nagwa Mostafa; Younis, Mohamed Saad; Elhamshary, Azza Mohamed; Abd-Elmaboud, Amina Ibrahim; Kishik, Shereen Magdy

    2014-09-01

    This study evaluated the performance of direct amplification of Acanthamoeba-DNA bypassing DNA extraction in the diagnosis of Acanthamoeba keratitis in clinically suspected cases in comparison to direct microscopic examination and in vitro culture. Corneal scrapings were collected from 110 patients who were clinically suspected to have Acanthamoeba keratitis, 63 contact lens wearers (CLW), and 47 non-contact lens wearers (NCLW). Taken samples were subjected to direct microscopic examination, cultivation onto the non-nutrient agar plate surface seeded with Escherichia coli, and PCR amplification. The diagnostic performance of these methods was statistically compared. The results showed that Acanthamoeba infection was detected in 21 (19.1%) of clinically suspected cases (110); 17 (81%) of them were CLW and the remaining 4 (19%) positive cases were NCLW. Regarding the used diagnostic methods, it was found that direct amplification of Acanthamoeba DNA bypassing nucleic acid extraction was superior to microscopy and culture in which 21 cases (19.1%) were positive for Acanthamoeba by PCR compared to 19 positive cases by culture (17.3%) and one case (0.9%) by direct smear. The difference in detection rates between culture and direct smear was highly statistically significant (P = 0.001). On the other hand, there was no significant difference in detection rates between culture and PCR (P = 0.86). On using culture as the gold standard, PCR showed three false-positive samples that were negative by culture and one false-negative sample that was positive by culture. At the same time, direct smear showed 18 false-negative samples. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of PCR were 94.7, 96.7, 85.7, 98.9, and 96.4, respectively, while those of direct smear were 5.3, 100, 100, 83.5, and 83.6, respectively. In conclusion, direct amplification of Acanthamoeba-DNA bypassing DNA extraction is a reliable

  15. Survival of environmental mycobacteria in Acanthamoeba polyphaga.

    Science.gov (United States)

    Adékambi, Toïdi; Ben Salah, Skandar; Khlif, Mohamed; Raoult, Didier; Drancourt, Michel

    2006-09-01

    Free-living amoebae in water are hosts to many bacterial species living in such an environment. Such an association enables bacteria to select virulence factors and survive in adverse conditions. Waterborne mycobacteria (WBM) are important sources of community- and hospital-acquired outbreaks of nontuberculosis mycobacterial infections. However, the interactions between WBM and free-living amoebae in water have been demonstrated for only few Mycobacterium spp. We investigated the ability of a number (n = 26) of Mycobacterium spp. to survive in the trophozoites and cysts of Acanthamoeba polyphaga. All the species tested entered the trophozoites of A. polyphaga and survived at this location over a period of 5 days. Moreover, all Mycobacterium spp. survived inside cysts for a period of 15 days. Intracellular Mycobacterium spp. within amoeba cysts survived when exposed to free chlorine (15 mg/liter) for 24 h. These data document the interactions between free-living amoebae and the majority of waterborne Mycobacterium spp. Further studies are required to examine the effects of various germicidal agents on the survival of WBM in an aquatic environment.

  16. Interaction between Vibrio mimicus and Acanthamoeba castellanii.

    Science.gov (United States)

    Abd, Hadi; Valeru, Soni Priya; Sami, Susan Marouf; Saeed, Amir; Raychaudhuri, Saumya; Sandström, Gunnar

    2010-02-01

    Vibrio mimicus is a Gram-negative bacterium, which causes gastroenteritis and is closely related to Vibrio cholerae. The environmental reservoir of this bacterium is far from defined. Acanthamoeba as well as Vibrio species are found in diverse aquatic environments. The present study was aimed to investigate the ability of A. castellanii to host V. mimicus, the role of bacterial protease on interaction with A. castellanii and to disclose the ability of cysts to protect intracellular V. mimicus. Co-cultivation, viable counts, gentamicin assay, electron microscopy and statistical analysis showed that co-cultivation of wild type and luxO mutant of V. mimicus strains with A. castellanii did not inhibit growth of the amoeba. On the other hand co-cultivation enhanced growth and survival of V. mimicus strains. Vibrio mimicus showed intracellular behaviour because bacteria were found to be localized in the cytoplasm of amoeba trophozoites and remain viable for 14 days. The cysts protected intracellular V. mimicus from high level of gentamicin. The intracellular growth of V. mimicus in A. castellanii suggests a role of A. castellanii as a host for V. mimicus.

  17. Lobectomía por VATS Video-thoracoscopic lobectomy

    OpenAIRE

    Patricio Rodríguez D; Felipe Undurraga M; Raimundo Santolaya C; David Lazo P; Jacqueline López R

    2012-01-01

    Introducción: En las últimas dos décadas los avances en cirugía mínimamente invasiva han revolucionado la práctica quirúrgica en todas las subespecialidades. En resecciones pulmonares la lobectomía por videotoracoscopía (VATS), en comparación con resecciones por toracotomía, ha demostrado, además del menor número de complicaciones perioperatorias, una menor respuesta inflamatoria y equivalencia en resultados oncológicos. Pacientes y Métodos: Se presenta una serie retrospectiva de 30 pacientes...

  18. In vitro Effect of Monosaccharides on the Virulence of Acanthamoeba Isolated from Patients with Amoebic Keratitis

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    Y. Maroofi

    2007-04-01

    Full Text Available Introduction & Objective: Acanthamoeba is free-living amoeba that is found in soil, water, air as well as in human pharynx. Acanthamoeba is causative agent of granulomatose amoebic encephalitis (GAE in immunosuppressed and AIDS individuals and amoebic keratitis in people who use the lens. Pathogenic species of Acanthamoeba have protein receptors named mannose binding protein (MBP. Acanthamoeba via MBP adhere to the glycoproteins included mannose. Acanthamoeba adhesion to the target cells induces a protease secretion is called mannose inducing protein-133 (MIP-133. Exogense mannose can inhibit the adherence of Acanthamoeba; also, it can increase the cytopathatic effect (CPE through increase the secretion of MIP-133. In the present work, the effect of monosaccharides on the virulance of Acanthamoeba isolated from patient with amoebic keratitis, in HeLa cell culture was investigated.Materials & Methods: The isolates were cultured in HeLa cell culture, then 100, 50, 10, 1 and 0.1 mM of galactose, glucose and mannose were added to plates. Plates were observed with invert microscope in 8, 16, 32, 48, and 72 hours after culture.Results: Data implicated that mannose (100 mM showed the highest effect on increasing cytopathy of Acanthamoeba in HeLa cell culture. Meanwhile, galactose (100 mM increased the virulence of Acanthamoeba in the cell culture after 32 hours.Conclusion: Adding mannose and galactose to HeLa cell culture contain Acanthamoeba can increase the virulence of the parasite significantly.

  19. Epidemiological typing of Acanthamoeba strains isolated from keratitis cases in Belgium.

    Science.gov (United States)

    De Jonckheere, J F

    2003-01-01

    From the corneas of nine keratitis patients and from their contact lenses, contact lens boxes and saline solutions, 15 strains of Acanthamoeba have been isolated. An Acanthamoeba strain was isolated from the swimming pool where one of the patients swam, while in the tapwater of the houses of three patients investigated, no Acanthamoeba could be detected. All the Acanthamoeba isolates from the cornea belong to genotype T4, but are different subtypes of T4. The Acanthamoeba detected on the contact lenses (and/or associated paraphernalia) of a patient are of the same subtype as that isolated from the cornea. The only Acanthamoeba strain isolated from a contact lens which was not related to an Acanthamoeba keratitis infection proved to be another genotype. A strain of Hartmannella from a cornea and two vahlkampfiids isolated from contact lenses had no connection with keratitis. This study confirms that, as found elsewhere, only Acanthamoeba genotype T4 of the 12 known Acanthamoeba genotypes is responsible for keratitis in Belgium. Most cases of Acanthamoeba keratitis cases are due to poor hygiene in the treatment (cleaning and storage) of contact lenses.

  20. Rapid and sensitive diagnosis of Acanthamoeba keratitis by loop-mediated isothermal amplification.

    Science.gov (United States)

    Ge, Z; Qing, Y; Zicheng, S; Shiying, S

    2013-11-01

    A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of Acanthamoeba. The sensitivity of the LAMP assay was tested using different copies of positive DNA. The specificity of the assay was tested using DNA extracted from Acanthamoeba, Pseudomonas aeruginosa, Candida albicans, herpes simplex virus-1 and human corneal epithelial cells. Its effectiveness was evaluated and compared with culture, corneal smear examination and real-time PCR in corneal samples from mice with Acanthamoeba keratitis. We also tested three corneal samples from patients with suspected Acanthamoeba or fungal infection using LAMP. Loop-mediated isothermal amplification was confirmed to be very sensitive, with the lowest detection limit being ten copies/tube of Acanthamoeba DNA. The LAMP primers only amplified Acanthamoeba DNA. During the development of Acanthamoeba keratitis in mice, almost all of the positive rates of LAMP at each time post-infection were higher than those of culture or corneal smear examination. The total positive rate of LAMP was significantly higher than those of culture and corneal smear examination (p Acanthamoeba keratitis tested positive for Acanthamoeba using LAMP along with culture or corneal smear examination, whereas the other suspected fungal keratitis tested negative. The LAMP assay is a simple, rapid, highly specific and sensitive method for the diagnosis of keratitis caused by Acanthamoeba.

  1. Tratamiento cognitivo-conductual protocolizado en grupo de las enfermedades inflamatorias intestinales Protocolized cognitive-behavioural group therapy for inflammatory bowel disease

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    M. A. Díaz Sibaja

    2007-10-01

    Full Text Available Introducción: los pacientes con enfermedades inflamatorias intestinales presentan una mayor tasa de alteraciones psicopatológicas que la población normal, predominantemente ansiedad y depresión. Objetivo: el principal objetivo de este estudio fue diseñar, poner en práctica y validar un programa de tratamiento psicológico protocolizado, basado en el modelo de competencias y con una metodología psicoeducativa, en el que se incluyeron los siguientes módulos de intervención: información de la enfermedad, modelo de afrontamiento, solución de problemas, técnicas de relajación, entrenamiento en habilidades sociales, técnicas de distracción y técnicas de reestructuración cognitiva. Todas estás técnicas fueron adaptadas a la problemática característica de la enfermedad inflamatoria intestinal. Método: la muestra estuvo constituida por 57 pacientes diagnosticados de enfermedad inflamatoria intestinal de la Asociación de Enfermos de Crohn y Colitis Ulcerosa de España. La muestra se distribuyó aleatoriamente en dos grupos experimentales, 33 formaron parte del programa de tratamiento en grupo y 24 fueron asignados al grupo control en lista de espera. Las variables emocionales medidas fueron: ansiedad y depresión. Resultados: los resultados reflejaron una mejoría clínica y estadísticamente significativa en las variables de ansiedad (p Introduction: patients affected by inflammatory bowel disease have a high rate of psychiatric disorders such as depression and anxiety. Objective: the main goal of this study was to design, to put into practice, and to validate a protocolized psychological treatment program based on the coping model, and a psycho-educational methodology in which the following intervention modules were included- illness information, coping model, problem solving techniques, relaxation, social skill training, distraction, and cognitive restructuring techniques. All these techniques were adapted to the characteristic

  2. Vaccination of patients with inflammatory bowel disease: Practical recommendations Vacunación de pacientes con enfermedad inflamatoria intestinal: Recomendaciones prácticas

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    Magda Campins

    2013-02-01

    Full Text Available Patients with inflammatory bowel disease (IBD have a greater risk of infection associated with the endogenous immunosuppression brought about by their underlying disease as well as the exogenous immunosuppression resulting from their therapies. In the last few years guidelines and consensus papers have been issued on the indication of vaccines for these patients as primary prevention of infection. However, vaccine coverage is low, likely because the risk for infection and both safety and efficacy concerns regarding vaccines are scarcely perceived in such cases. The available scientific evidence shows that immunization is safe for most preparations, and bears no association with an increased risk for disease activity. This paper reviews the available scientific literature, and provides recommendations on the vaccination of adults with IBD.Los pacientes con enfermedad inflamatoria intestinal (EII tienen un mayor riesgo de infecciones, asociado tanto a la inmunosupresión endógena condicionada por su enfermedad de base, como a la exógena generada por los tratamientos que reciben. En los últimos años se han publicado guías y documentos de consenso sobre indicaciones de vacunación en estos pacientes, como medida de prevención primaria de infecciones. Sin embargo, las coberturas vacunales alcanzadas son bajas, probablemente por la falta de percepción del riesgo de infección y las dudas sobre la seguridad y eficacia de las vacunas en estos casos. La evidencia científica disponible muestra que la inmunización es segura para la mayoría de preparados y no se asocia a un incremento del riesgo de actividad de la enfermedad. En este documento se revisa la literatura científica disponible y se presentan unas recomendaciones de vacunación para pacientes adultos con EII.

  3. Citocinas anti-inflamatorias y sus acciones y efectos en la sepsis y el choque séptico - Anti-inflammatory cytokines and their actions and effects in the sepsis and septic shock

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    González Álvarez, Ricardo

    2009-09-01

    Full Text Available ResumenLas citocinas anti- inflamatorias reconocidas son las interleucinas (ILIL-4, IL-10, IL-6, IL-13, el factor estimulante de colonia granulocito – macrófago (FEC-GM y el interferón alfa (IFN-alfa, los cuales tienen la capacidad de inhibir la liberación de las citocinas pro-inflamatorias y de inducir la producción del antagonista del receptor de la IL-1 y la liberación del receptor soluble del FNT, los cuales limitan algunas de las actividades de las citocinas pro-inflamatorias IL-1 y FNT-α. Sin embargo, los eventos que ocurren durante la inflamación y la sepsis no son tan simples como para justificar siempre acciones antagonistas entre las citocinas pro-inflamatorias y las antiinflamatorias, las cuales, pueden tener acorde con las circunstancias un comportamiento dual y/o antagónico en el choque séptico. Así por ejemplo la IL-10 tiene propiedades anti-inflamatorias, lo cual ocurre en la diabetes auto-inmune cuyo comienzo y desarrollo son acelerados en ratones transgénicos que sobre-expresan la IL-10 en los islotes pancreáticos. Esto también ocurre con la IL-10 en un modelo de uveítis en el que se incrementó la inflamación ocular. Ejemplos como los anteriores son extensivos a otras citocinas antiinflamatorias como la IL-4, IL-13, el factor estimulante de colonia granulocito –macrófago y el factor beta transformante del crecimiento (FβTC, y que serán mencionadas y descritas en el presente artículo de revisión, así como los factores que determinan ese comportamiento dual de las citocinas.SummaryThe anti-inflammatory cytokines which are recognized are theinterleukins (IL IL-4, IL-10, IL-13, interferon-alpha (IFN-α andbecause they are able to inhibit the release of pro inflammatorycytokines, to induce the production of IL-1 receptor antagonist (IL-raand the release of soluble TNF receptor and to limit some of the proinflammatory activities of IL-1 and TNF. However the events whichoccur during inflammation and sepsis are

  4. A retrospective study of nine cases of Acanthamoeba keratitis

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    Tetsuya Mutoh

    2010-10-01

    Full Text Available Tetsuya Mutoh, Isao Ishikawa, Yukihiro Matsumoto, Makoto ChikudaDokkyo Medical University Koshigaya Hospital, Saitama, JapanPurpose: To evaluate the clinical features of Acanthamoeba keratitis in nine patients diagnosed at Dokkyo Medical University Koshigaya Hospital, Saitama, Japan.Methods: In nine eyes of nine patients, Acanthamoeba keratitis was diagnosed by direct light microscopy of corneal scrapings stained by the Parker ink-potassium hydroxide method between September 2006 and September 2009. Their clinical features and course were studied retrospectively. Antifungal eye drops, systemic antifungal therapy, and surgical debridement of the corneal lesions were performed in all patients.Results: At presentation, the clinical stage was initial in six cases, transient in one case, and complete in two cases. The patients were all contact lens wearers who had washed their lens storage cases with tap water. After treatment, final visual acuity was improved in six cases, unchanged in one case, and worse in two cases. The patient with the worst final vision (hand motion had rheumatoid arthritis and was taking oral prednisolone, which led to corneal perforation and prevented adequate debridement from being done.Conclusion: Acanthamoeba keratitis is closely related to wearing contact lenses and washing the lens storage case with tap water. Although final visual acuity improved after treatment in most patients, insufficient surgical debridement resulted in a poor visual prognosis.Keywords: surgical debridement, Acanthamoeba keratitis, contact lens wearers

  5. Cytotoxic activities of alkylphosphocholines against clinical isolates of Acanthamoeba spp.

    Science.gov (United States)

    Walochnik, Julia; Duchêne, Michael; Seifert, Karin; Obwaller, Andreas; Hottkowitz, Thomas; Wiedermann, Gerhard; Eibl, Hansjörg; Aspöck, Horst

    2002-03-01

    Free-living amoebae of the genus Acanthamoeba are causing serious chronic conditions such as destructive keratitis in contact lens wearers or granulomatous amoebic encephalitis in individuals with compromised immune systems. Both are characterized by the lack of availability of sufficiently effective and uncomplicated, manageable treatments. Hexadecylphosphocholine (miltefosine) is licensed for use as a topical antineoplastic agent, but it is also active in vitro against several protozoan parasites, and it was applied very successfully for the treatment of human visceral leishmaniasis. The aim of our study was to evaluate the efficacy of hexadecylphosphocholine and other alkylphosphocholines (APCs) against Acanthamoeba spp. The in vitro activities of eight different APCs against three Acanthamoeba strains of various pathogenicities were determined. All substances showed at least amoebostatic effects, and some of them disrupted the amoebae, as shown by the release of cytoplasmic enzyme activity. Hexadecylphosphocholine exhibited the highest degree of cytotoxicity against trophozoites, resulting in complete cell death at a concentration as low as 40 microM, and also displayed significant cysticidal activity. Hexadecylphosphocholine may be a promising new candidate for the topical treatment of Acanthamoeba keratitis and, conceivably, even for the oral treatment of granulomatous amoebic encephalitis.

  6. three cases of acanthamoeba keratitis (Ak in Yazd

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    MR Shoja

    2006-04-01

    Full Text Available Acanthamoeba keratitis is a vision- threatening infection and the most important risk factors are contact lenses Characteristic symptoms include disproportionately severe ocular pain, epithelial ulcer, ring stromal infiltrate and resistance to antimicrobial treatment. The aim is to report the first cases of Acanthamoeba keratitis in Yazd in a retrospective study confirmed on histopathological and confocal microscopy. All of our patients used soft contact lenses and were women. The age range was between 20-29 years. One eye was`misdiagnosed as herpes simplex keratitis and two were misdiagnosed as bacterial keratitis. A delayed visit as well as a late diagnosis led to the advanced stage of eye disease. Improvement in visual acuity following medical therapy was seen in two patients and one patient needed urgent keratoplasty for corneal necrosis A prompt clinical and laboratory diagnosis along with treatment with propamidine- isentionate resulted in significant improvement of eye condition Failure to disinfect contact lenses and introduction of tap water rinsing of storage cases were associated with increased risk of Acanthamoeba infection. Penetrating keratoplasty is an effective treatment for medically unresponsive acanthamoeba keratitis. Confocal microscopy is the most effective and a fast diagnostic tool because it ensures the detection of acanthamebic cysts and trophozoids in corneal stroma .

  7. Colonization of broilers by Campylobacter jejuni internalized within Acanthamoeba castellanii

    Science.gov (United States)

    We present the first report that Campylobacter jejuni, internalized within Acanthamoeba castellanii, colonized broilers. After 1, 3, 7 and 14 days post challenge none of the broilers challenged with negative controls were colonized, but were with internalized C. jejuni. The biology of protozoa-Cam...

  8. Acanthamoeba polyphaga mimivirus virophage seroconversion in travelers returning from Laos.

    Science.gov (United States)

    Parola, Philippe; Renvoisé, Aurélie; Botelho-Nevers, Elisabeth; La Scola, Bernard; Desnues, Christelle; Raoult, Didier

    2012-09-01

    During January 2010, a husband and wife returned from Laos to France with probable parasitic disease. Increased antibodies against an Acanthamoeba polyphaga mimivirus virophage indicated seroconversion. While in Laos, they had eaten raw fish, a potential source of the virophage. This virophage, associated with giant viruses suspected to cause pneumonia, could be an emerging pathogen.

  9. Acanthamoeba polyphaga in rheumatoid arthritis: possibility for a chronic infection.

    Science.gov (United States)

    Jeansson, S; Kvien, T K

    2001-06-01

    Acanthamoeba polyphaga (AP) is ubiquitous in nature and frequently infects humans. AP has some features, such as persistence, which makes it an attractive candidate in studies of a possible infectious aetiology in rheumatoid arthritis (RA). In this study the occurrence of AP-specific antibodies was compared between RA patients and matched controls.

  10. Identification of unusual phospholipid fatty acyl compositions of Acanthamoeba castellanii.

    Science.gov (United States)

    Palusinska-Szysz, Marta; Kania, Magdalena; Turska-Szewczuk, Anna; Danikiewicz, Witold; Russa, Ryszard; Fuchs, Beate

    2014-01-01

    Acanthamoeba are opportunistic protozoan pathogens that may lead to sight-threatening keratitis and fatal granulomatous encephalitis. The successful prognosis requires early diagnosis and differentiation of pathogenic Acanthamoeba followed by aggressive treatment regimen. The plasma membrane of Acanthamoeba consists of 25% phospholipids (PL). The presence of C20 and, recently reported, 28- and 30-carbon fatty acyl residues is characteristic of amoeba PL. A detailed knowledge about this unusual PL composition could help to differentiate Acanthamoeba from other parasites, e.g. bacteria and develop more efficient treatment strategies. Therefore, the detailed PL composition of Acanthamoeba castellanii was investigated by 31P nuclear magnetic resonance spectroscopy, thin-layer chromatography, gas chromatography, high performance liquid chromatography and liquid chromatography-mass spectrometry. Normal and reversed phase liquid chromatography coupled with mass spectrometric detection was used for detailed characterization of the fatty acyl composition of each detected PL. The most abundant fatty acyl residues in each PL class were octadecanoyl (18∶0), octadecenoyl (18∶1 Δ9) and hexadecanoyl (16∶0). However, some selected PLs contained also very long fatty acyl chains: the presence of 28- and 30-carbon fatty acyl residues was confirmed in phosphatidylethanolamine (PE), phosphatidylserine, phosphatidic acid and cardiolipin. The majority of these fatty acyl residues were also identified in PE that resulted in the following composition: 28∶1/20∶2, 30∶2/18∶1, 28∶0/20∶2, 30∶2/20∶4 and 30∶3/20∶3. The PL of amoebae are significantly different in comparison to other cells: we describe here for the first time unusual, very long chain fatty acids with Δ5-unsaturation (30∶35,21,24) and 30∶221,24 localized exclusively in specific phospholipid classes of A. castellanii protozoa that could serve as specific biomarkers for the presence of these

  11. Identification of unusual phospholipid fatty acyl compositions of Acanthamoeba castellanii.

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    Marta Palusinska-Szysz

    Full Text Available Acanthamoeba are opportunistic protozoan pathogens that may lead to sight-threatening keratitis and fatal granulomatous encephalitis. The successful prognosis requires early diagnosis and differentiation of pathogenic Acanthamoeba followed by aggressive treatment regimen. The plasma membrane of Acanthamoeba consists of 25% phospholipids (PL. The presence of C20 and, recently reported, 28- and 30-carbon fatty acyl residues is characteristic of amoeba PL. A detailed knowledge about this unusual PL composition could help to differentiate Acanthamoeba from other parasites, e.g. bacteria and develop more efficient treatment strategies. Therefore, the detailed PL composition of Acanthamoeba castellanii was investigated by 31P nuclear magnetic resonance spectroscopy, thin-layer chromatography, gas chromatography, high performance liquid chromatography and liquid chromatography-mass spectrometry. Normal and reversed phase liquid chromatography coupled with mass spectrometric detection was used for detailed characterization of the fatty acyl composition of each detected PL. The most abundant fatty acyl residues in each PL class were octadecanoyl (18∶0, octadecenoyl (18∶1 Δ9 and hexadecanoyl (16∶0. However, some selected PLs contained also very long fatty acyl chains: the presence of 28- and 30-carbon fatty acyl residues was confirmed in phosphatidylethanolamine (PE, phosphatidylserine, phosphatidic acid and cardiolipin. The majority of these fatty acyl residues were also identified in PE that resulted in the following composition: 28∶1/20∶2, 30∶2/18∶1, 28∶0/20∶2, 30∶2/20∶4 and 30∶3/20∶3. The PL of amoebae are significantly different in comparison to other cells: we describe here for the first time unusual, very long chain fatty acids with Δ5-unsaturation (30∶35,21,24 and 30∶221,24 localized exclusively in specific phospholipid classes of A. castellanii protozoa that could serve as specific biomarkers for the presence of

  12. Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays.

    Science.gov (United States)

    Khan, N A; Jarroll, E L; Paget, T A

    2001-09-01

    Acanthamoeba are opportunistic pathogens with invasive and noninvasive species. For clinical purposes it is important to differentiate potentially pathogenic from nonpathogenic isolates. For the rapid and sensitive identification of Acanthamoeba at the genus level, we used a polymerase chain reaction (PCR)-based method which detected as few as five cells. Further, we tested nine isolates of Acanthamoeba for their ability to produce cytopathic effects (CPE) on corneal epithelial cells. On the basis of the results, Acanthamoeba were divided into pathogenic or nonpathogenic groups. However, because CPE assays are not available to every diagnostic laboratory, we developed a simple plating assay based on osmotolerance which correlated well with the CPE assays. Pathogenic Acanthamoeba showed growth on higher osmolarity (agar plates containing one molar mannitol), while growth of nonpathogens was inhibited on these plates. In conclusion, we have developed methods for the rapid identification and differentiation of Acanthamoeba.

  13. Acanthamoeba keratitis - A six year epidemiological review from a tertiary care eye hospital in South India

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    Manikandan P

    2004-01-01

    Full Text Available PURPOSE: This study analyses the prevalence, demography, predisposing factors and seasonal variation of Acanthamoeba keratitis. METHODS: A retrospective review of all cases presenting with keratitis at the cornea clinic, Aravind Eye Hospital, Coimbatore, from August 1997 to July 2003, was done for screening patients with a provisional diagnosis of Acanthamoeba keratitis. Their records were further analyzed for microbiological details. Cases with culture proven Acanthamoeba keratitis were included for epidemiological analysis. RESULTS: From a total of 4519 patients who attended cornea clinic 32 (33 eyes patients were confirmed to be positive for Acanthamoeba keratitis. Twenty cases (62.5% were males. Majority (18; 54.2% of the Acanthamoeba keratitis eyes reported corneal trauma by solid objects. No peak period was observed in a year, as the number of cases was almost uniform in all months. CONCLUSION: This study indicates the increasing prevalence of Acanthamoeba keratitis among non-contact lens users in this region during the 6-year period.

  14. Molecular detection and genotyping of Acanthamoeba spp. among stray dogs using conjunctival swab sampling.

    Science.gov (United States)

    Karakuş, Mehmet; Aykur, Mehmet; Özbel, Yusuf; Töz, Seray; Dağcı, Hande

    2016-12-01

    Acanthamoeba is one of the most common free-living amoebas (FLA) that present in environment. In humans, Acanthamoeba can cause an infection of the eye termed Acanthamoeba keratitis, which mostly occurs in contact lens wearers. In the present study, we aimed to screen the presence of Acanthamoeba DNA in stray dogs using previously collected conjunctival swab samples in a hyper-endemic area for canine leishmaniasis. Totally, 184 dogs were included in the study and 27 of them (14.6%) were found positive for Acanthamoeba according to the 18s rRNA gene sequencing. Two different genotypes (T4 and T5) were identified and T5 was firstly reported in Turkey in the present study. Statistical analysis was performed and no correlation was found between Leishmania and Acanthamoeba positivity (PAcanthamoeba among stray dogs. Further studies are necessary to reveal the infection status and genotypes among dogs and its possible correlation with leishmaniasis.

  15. A method for microbial decontamination ofAcanthamoeba cultures using the peritoneal cavity of mice

    Institute of Scientific and Technical Information of China (English)

    Rodrigo Gurgel-Gonalves; Patrcia Albuquerque; Csar Augusto Cuba-Cuba

    2015-01-01

    Objective:To evaluate whether the inoculation of contaminated cultures in the peritoneal cavity of mice could implement decontamination ofAcanthamoeba cultures. Methods: Suspensions ofAcanthamoeba,Acanthamoeba polyphagaATCC30461, or Acanthamoeba spp. isolated from soil (UnB13 strain) were inoculated in the peritoneal cavity of Swiss mice (n = 24). After 1, 6, 12, or 24 h of exposure the peritoneal cavity was washed and assessed for the presence of bacteria, fungi, andAcanthamoeba. Results: After 1 h of intraperitoneal inoculation at least 97% of the bacteria and 96% of the fungi (P Conclusions: Our data demonstrated that this technique has great potential for decontamination ofAcanthamoeba cultures in a short period of time.

  16. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii Trophozoites and Cysts

    OpenAIRE

    Matin, Abdul; Jung, Suk-Yul

    2011-01-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. ...

  17. Diversity and Seasonal Impact of Acanthamoeba Species in a Subtropical Rivershed

    OpenAIRE

    Po-Min Kao; Ming-Yuan Chou; Chi-Wei Tao; Wen-Chien Huang; Bing-Mu Hsu; Shu-Min Shen; Cheng-Wei Fan; Yi-Chou Chiu

    2013-01-01

    This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in th...

  18. Acanthamoeba castellanii an environmental host for Shigella dysenteriae and Shigella sonnei.

    Science.gov (United States)

    Saeed, Amir; Abd, Hadi; Edvinsson, Benjamin; Sandström, Gunnar

    2009-01-01

    The interaction between Shigella dysenteriae or Shigella sonnei and Acanthamoeba castellanii was studied by viable counts, gentamicin assay and electron microscopy. The result showed that Shigella dysenteriae or Shigella sonnei grew and survived in the presence of amoebae for more than 3 weeks. Gentamicin assay showed that the Shigella were viable inside the Acanthamoeba castellanii which was confirmed by electron microscopy that showed the Shigella localized in the cytoplasm of the Acanthamoeba castellanii. In conclusion, the relationship between Shigella dysenteriae and Shigella sonnei with Acanthamoeba castellanii is symbiotic, and accordingly free-living amoebae may serve as a transmission reservoir for Shigella in water.

  19. The role of the innate and adaptive immune responses in Acanthamoeba keratitis.

    Science.gov (United States)

    Niederkorn, Jerry Y

    2002-01-01

    Infections of the corneal surface are an important cause of blindness. Protozoal, viral, bacterial, and helminthic infections of the cornea account for up to 9 million cases of corneal blindness. Free-living amoebae of the genus Acanthamoeba produce a progressive infection of the cornea called Acanthamoeba keratitis. Disease is usually transmitted by Acanthamoeba trophozoites bound to soft contact lenses. Infection of the cornea is initiated when the parasite binds to the corneal epithelial surface. Recrudescence can occur and suggests that the adaptive immune response is not aroused by corneal Acanthamoeba infections. Systemic immunization with Acanthamoeba antigens elicits robust Th1 cell-mediated immunity and serum IgG antibody, yet fails to prevent the development of Acanthamoeba keratitis. However, immunization via mucosal surfaces induces anti-Acanthamoeba IgA antibodies in the tears and provides solid protection against the development of Acanthamoeba keratitis. Unlike other immune effector mechanisms that rely on cytolysis, inflammation, release of toxic molecules, or the induction of host cell death, the adaptive immune apparatus prevents Acanthamoeba infections of the cornea by simply preventing the attachment of the parasite to the epithelial surface. The beauty of this mechanism lies in its exquisite simplicity and efficacy.

  20. Acanthamoeba protease activity promotes allergic airway inflammation via protease-activated receptor 2.

    Science.gov (United States)

    Park, Mi Kyung; Cho, Min Kyoung; Kang, Shin Ae; Park, Hye-Kyung; Kim, Dong-Hee; Yu, Hak Sun

    2014-01-01

    Acanthamoeba is a free-living amoeba commonly present in the environment and often found in human airway cavities. Acanthamoeba possesses strong proteases that can elicit allergic airway inflammation. To our knowledge, the aeroallergenicity of Acanthamoeba has not been reported. We repeatedly inoculated mice with Acanthamoeba trophozoites or excretory-secretory (ES) proteins intra-nasally and evaluated symptoms and airway immune responses. Acanthamoeba trophozoites or ES proteins elicited immune responses in mice that resembled allergic airway inflammation. ES proteins had strong protease activity and activated the expression of several chemokine genes (CCL11, CCL17, CCL22, TSLP, and IL-25) in mouse lung epithelial cells. The serine protease inhibitor phenyl-methane-sulfonyl fluoride (PMSF) inhibited ES protein activity. ES proteins also stimulated dendritic cells and enhanced the differentiation of naive T cells into IL-4-secreting T cells. After repeated inoculation of the protease-activated receptor 2 knockout mouse with ES proteins, airway inflammation and Th2 immune responses were markedly reduced, but not to basal levels. Furthermore, asthma patients had higher Acanthamoeba-specific IgE titers than healthy controls and we found Acanthamoeba specific antigen from house dust in typical living room. Our findings suggest that Acanthamoeba elicits allergic airway symptoms in mice via a protease allergen. In addition, it is possible that Acanthamoeba may be one of the triggers human airway allergic disease.

  1. Characterization of isolates of Acanthamoeba from the nasal mucosa and cutaneous lesions of dogs.

    Science.gov (United States)

    Carlesso, A M; Mentz, M B; da Machado, M L S; Carvalho, A; Nunes, T E T; Maschio, V J; Rott, M B

    2014-06-01

    Acanthamoeba spp. are free-living amoebae that are ubiquitously distributed in the environment and can cause encephalomyelitis in animals and humans. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread, and host susceptibility. The aim of the present study was to characterize isolates of Acanthamoeba from the nasal mucosa and cutaneous lesions of dogs in order to access the occurence and pathogenicity of these organisms in this animal group. We studied 13 isolates of Acanthamoeba confirmed by polymerase chain reaction. They were sequenced, the genotype was determined, and their potential of pathogenicity was evaluated.

  2. Seasonal distribution of potentially pathogenic Acanthamoeba species from drinking water reservoirs in Taiwan.

    Science.gov (United States)

    Kao, Po-Min; Hsu, Bing-Mu; Hsu, Tsui-Kang; Liu, Jorn-Hon; Chang, Hsiang-Yu; Ji, Wen-Tsai; Tzeng, Kai-Jiun; Huang, Shih-Wei; Huang, Yu-Li

    2015-03-01

    In order to detect the presence/absence of Acanthamoeba along with geographical variations, water quality variations and seasonal change of Acanthamoeba in Taiwan was investigated by 18S ribosomal RNA (rRNA) gene TaqMan quantitative real-time PCR. Samples were collected quarterly at 19 drinking water reservoir sites from November 2012 to August 2013. Acanthamoeba was detected in 39.5 % (30/76) of the water sample, and the detection rate was 63.2 % (12/19) from samples collected in autumn. The average concentration of Acanthamoeba was 3.59 × 10(4) copies/L. For geographic distribution, the detection rate for Acanthamoeba at the northern region was higher than the central and southern regions in all seasons. Results of Spearman rank test revealed that heterotrophic plate count (HPC) had a negative correlation (R = -0.502), while dissolved oxygen (DO) had a positive correlation (R = 0.463) in summer. Significant differences were found only between the presence/absence of Acanthamoeba and HPC in summer (Mann-Whitney U test, P Acanthamoeba were identified, and T4 was the most commonly identified Acanthamoeba genotypes. The presence of Acanthamoeba in reservoirs presented a potential public health threat and should be further examined.

  3. Acanthamoeba protease activity promotes allergic airway inflammation via protease-activated receptor 2.

    Directory of Open Access Journals (Sweden)

    Mi Kyung Park

    Full Text Available Acanthamoeba is a free-living amoeba commonly present in the environment and often found in human airway cavities. Acanthamoeba possesses strong proteases that can elicit allergic airway inflammation. To our knowledge, the aeroallergenicity of Acanthamoeba has not been reported. We repeatedly inoculated mice with Acanthamoeba trophozoites or excretory-secretory (ES proteins intra-nasally and evaluated symptoms and airway immune responses. Acanthamoeba trophozoites or ES proteins elicited immune responses in mice that resembled allergic airway inflammation. ES proteins had strong protease activity and activated the expression of several chemokine genes (CCL11, CCL17, CCL22, TSLP, and IL-25 in mouse lung epithelial cells. The serine protease inhibitor phenyl-methane-sulfonyl fluoride (PMSF inhibited ES protein activity. ES proteins also stimulated dendritic cells and enhanced the differentiation of naive T cells into IL-4-secreting T cells. After repeated inoculation of the protease-activated receptor 2 knockout mouse with ES proteins, airway inflammation and Th2 immune responses were markedly reduced, but not to basal levels. Furthermore, asthma patients had higher Acanthamoeba-specific IgE titers than healthy controls and we found Acanthamoeba specific antigen from house dust in typical living room. Our findings suggest that Acanthamoeba elicits allergic airway symptoms in mice via a protease allergen. In addition, it is possible that Acanthamoeba may be one of the triggers human airway allergic disease.

  4. Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish

    OpenAIRE

    Im, Kyung-Il; Shin, Ho-Joon

    2003-01-01

    A new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Acanthamoeba sp. YM-4 (Korean isolate YM-4). The trophozoites were 11.0-23.0 µm in length and had hyaline filamentous projections. Cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Acanthamoeba YM-4 can survive at 40℃, and its generation time was 19.6 hr, which was longer than that of A. culbertsoni. In terms of the in vitro c...

  5. Thermotolerant Acanthamoeba spp. isolated from therapeutic hot springs in Northwestern Iran.

    Science.gov (United States)

    Solgi, Rahmat; Niyyati, Maryam; Haghighi, Ali; Taghipour, Niloofar; Tabaei, Seyyed Javad Seyyed; Eftekhar, Mohamad; Nazemalhosseini Mojarad, Ehsan

    2012-12-01

    This study was conducted to address the distribution of Acanthamoeba genotypes in therapeutic hot springs in Iran. Sixty water and sediment samples were collected from bicarbonate, sulphur, and sodium chloride thermal springs in the northwest. All hot springs examined are used mainly for health purposes in Iran. Acanthamoeba were identified by both morphology and PCR (polymerase chain reaction). Genotype identification was based on the sequencing of a highly variable and informative region of Diagnostic Fragment 3 (stem 29-1 of 18S rRNA gene) within Acanthamoeba-specific amplimer (ASA.S1). Twenty percent of hot springs were contaminated with thermotolerant Acanthamoeba belonging to the potentially pathogenic T4 and T3 genotypes. A high number (91.7%) of strains showed growth at 37 °C, and eight isolates showed growth at 42 °C. A single isolate (HSNW2) was detected in waters at 70 °C. The presence of thermotolerant Acanthamoeba highlights a risk factor for susceptible individuals, as Acanthamoeba-related keratitis continues to rise in Iran. Periodic surveillance of thermal waters as well as improved filtration and disinfection is recommended to prevent disease related to pathogenic Acanthamoeba. This is the first comprehensive molecular study of Acanthamoeba genotypes in hot springs in Iran and the first to report the occurrence of the T3 genotype (corresponding to Acanthamoeba griffini) in thermal water sources in this country.

  6. MASCULINO DE 55 AÑOS CON MENINGOENCEFALITIS POR FLAVIVIRUS, POLIRRADICULOPATÍA INFLAMATORIA Y EVC ISQUÉMICO

    OpenAIRE

    Quesada Aguilar, Carlos I.

    2011-01-01

    We present a young patient with untreated hypertension, who complained of acute loss of muscular strength on the left side of his body. He hadfever and the diagnostic work up showed lymphocytic meningitis. He also presented withgeneralized weakness in his four extremities anda neurophysiologic study showed an inflammato-ry poliradiculoneuropathy. The PCR analysis ofthe cerebrospinal fluid was positive for flavivirusinfection. Se describe el caso de un paciente masculino joven, hipertenso s...

  7. Estudio PET/TC en patología inflamatoria-infecciosa PET/CT in infectious and inflammatory pathology

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    Cecilia Carrera

    2012-06-01

    Full Text Available Objetivo. Demostrar la utilidad del examen PET/TC en patología inflamatoria-infecciosa. Materiales y Métodos. Evaluación retrospectiva (enero de 2009 - mayo de 2011 de los exámenes de tomografía por Emisión de Positrones/ Tomografía Computada (PET/TC, realizados en nuestra institución con un equipo híbrido SIEMENS-BIOGRAPH 16 (Siemens, Erlangen, Alemania. Se seleccionaron 5 pacientes. Resultados. Caso 1: paciente de 68 años de edad con fiebre de 6 meses de duración, fatiga y pérdida de peso. El examen reumatológico demostró disminución en pulsos radiales sin otros síntomas asociados. La paciente fue sometida a biopsia de arteria temporal. Ésta confirmó una arteritis de la arteria temporal y el estudio PET/TC demostró hipermetabolismo en la aorta torácica y ramas principales. Caso 2: paciente de 85 años con fiebre de origen desconocido (FOD y sospecha de osteomielitis de cadera. En contraposición, el PET/TC demostró un foco ávido de celulitis glútea y neumopatía. Caso 3: paciente de 35 años con fiebre vespertina. El PET/TC mostró múltiples adenomegalias ávidas por fluorodexosiglucosa (FDG en mediastino, axilas y retroperitoneo, y compromiso difuso esplénico asociado a calcificaciones. Se confirmó infección por citomegalovirus por inmuno-globulina G y M. Caso 4: paciente de 39 años con infección por HIV que consultó por hipercalcemia. El PET/TC mostró implantes de silicona en glúteos con proceso inflamatorio ávido asociado. Se confirmó por la biopsia de uno de ellos. Caso 5: paciente de 45 años con historia de cáncer de mama en control presentó en los últimos estudios tomográficos aumento del tamaño de los ganglios supraclaviculares y mediastínicos, y compromiso esplénico multifocal difuso. Estos resultaron ávidos en el examen PET/TC. Se confirmó el diagnóstico de sarcoidosis por el estudio anatomopatológico de un ganglio supraclavicular. Conclusiones. El PET/TC es un método no invasivo de

  8. Nutrición y enfermedad inflamatoria intestinal Nutrition and inflammatory bowel disease

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    E. Cabré Gelada

    2007-05-01

    Full Text Available En este artículo se revisan las causas y consecuencias de los déficits nutricionales en la enfermedad inflamatoria intestinal, con especial énfasis en sus implicaciones fisiopatológicas. Además, se sientan las bases de la intervención nutricional en estos pacientes tanto desde la perspectiva del consejo dietético como del uso de soporte nutricional especializado. Se hace un especial hincapié en la discusión de las evidencias existentes en favor de un papel terapéutico primario de la nutrición artificial y algunos nutrientes específicos en la enfermedad de Crohn.In this paper, the causes and consequences of nutritional deficiencies in inflammatory bowel disease are reviewed. Particular emphasis is made on their pathophysiological implications. In addition, the basis for nutritional intervention in these patients are described (both in terms of dietary counseling and the use of specialized nutritional support. Particular mention is made on the current evidences in favour of a primary therapeutic role of artificial nutrition and some specific nutrients in Crohn's disease.

  9. Molecular and physiological differentiation between pathogenic and nonpathogenic Acanthamoeba.

    Science.gov (United States)

    Khan, Naveed A; Jarroll, Edward L; Paget, Timothy A

    2002-09-01

    In this study, 14 isolates of Acanthamoeba from both clinical and environmental sources belonging to seven different species were assayed for tolerance of high osmotic pressure, temperature tolerance, extracellular proteases, and cytopathic effects (CPE) on immortalized rabbit corneal epithelial cells. On the basis of the results, amoeba isolates were divided into pathogenic and nonpathogenic groups. Ribosomal DNA sequencing was performed on these isolates. Phylogenetic relationships revealed that all the pathogenic strains tested clustered together as one group, while nonpathogenic strains clustered into other groups. Sequence comparisons with previously published sequences determined that among the six new pathogenic isolates used in this study, five belong to T4 genotype and one to T11. This is the first report of a T11 genotype being found in Acanthamoeba keratitis.

  10. Double-Biguanide Therapy for Resistant Acanthamoeba Keratitis

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    Giulio Ferrari

    2011-11-01

    Full Text Available Aims: To report the clinical and diagnostic findings of a patient with Acanthamoeba keratitis resistant to both polyhexamethylene biguanide (PHMB-hexamidine and chlorhexidine-hexamidine treatment. Methods: Slit-lamp biomicroscopy, corneal cell scraping and histopathology were performed on a 39-year-old woman presenting with corneal ulcer in her left eye. Results: The patient was successfully treated with PHMB-chlorhexidine association therapy. Subsequent perforating keratoplasty remained clear at the last follow-up visit after 7 months and increased visual acuity to 20/20 with correction. Conclusions: This case emphasizes the proteiform aspects of Acanthamoeba drug resistance, and suggests that PHMB-chlorhexidine association might represent an additional option for cases resistant to standard therapy.

  11. Detection of glycoproteins in the Acanthamoeba plasma membrane

    Energy Technology Data Exchange (ETDEWEB)

    Paatero, G.I.L. (Abo Akademi (Finland)); Gahmberg, C.G. (Univ. of Helsinki (Finland))

    1988-11-01

    In the present study the authors have shown that glycoproteins are present in the plasma membrane of Acanthamoeba castellanii by utilizing different radioactive labeling techniques. Plasma membrane proteins in the amoeba were iodinated by {sup 125}I-lactoperoxidase labeling and the solubilized radiolabeled glycoproteins were separated by lectin-Sepharose affinity chromatography followed by polyacrylamide gel electrophoresis. The periodate/NaB{sup 3}H{sub 4} and galactose oxidase/NaB{sup 3}H{sub 4} labeling techniques were used for labeling of surface carbohydrates in the amoeba. Several surface-labeled glycoproteins were observed in addition to a diffusely labeled region with M{sub r} of 55,000-75,000 seen on electrophoresis, which could represent glycolipids. The presence of glycoproteins in the plasma membrane of Acanthamoeba castellanii was confirmed by metabolic labeling with ({sup 35}S)methionine followed by lectin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis.

  12. O-Linked glycosylation in Acanthamoeba polyphaga mimivirus.

    Science.gov (United States)

    Hülsmeier, Andreas J; Hennet, Thierry

    2014-08-01

    Acanthamoeba polyphaga mimivirus is a member of the giant nucleocytoplasmic large DNA viruses, infecting various Acanthamoeba spp. The genomes of giant viruses encode components previously thought to be exclusive to cellular life, such as proteins involved in nucleic acid and protein synthesis. Recent work on enzymes involved in carbohydrate biosynthesis and metabolism show that instead of utilizing host cell resources, Mimivirus produces its own glycosylation machinery. To obtain a more detailed view of glycosylation in Mimivirus, we developed a periodate oxidation-based method to selectively enrich Mimivirus surface glycoproteins. O-Glycosylation in Mimivirus glycoproteins was identified by permethylation and matrix-assisted laser desorption/ionization-mass spectrometry analyses of beta-eliminated glycans. We sequenced 26 previously undescribed O-glycans, most of which contain glucose as their reducing end saccharide. These data will facilitate future studies on the functional significance of glycosylation in Mimivirus.

  13. Partial characterization of Acanthamoeba castellanii (T4 genotype) DNase activity.

    Science.gov (United States)

    Iqbal, Junaid; Panjwani, Shamvil; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2015-02-01

    The deoxyribonuclease (DNase) activities of Acanthamoeba castellanii belonging to the T4 genotype were investigated. Using zymographic assays, the DNase activities had approximate molecular masses of 25 and 35 kDa. A. castellanii DNases exhibited activity at wide-ranging temperature of up to 60 °C and at pH ranging from 4 to 9. The DNases activities were unaffected by proteinase-K treatment, divalent cations such as Ca(++), Cu(++), Mg(++), and Zn(++), or divalent cation chelating agent ethylenediaminetetraacetic acid (EDTA) or sodium dodecyl sulfate (SDS). The non-reliance on divalent cations and homology data suggests that A. castellanii DNases belong to the class of eukaryotic lysosomal DNase II but exhibit robust properties. The DNases activity in A. castellanii interfered with the genomic DNA extraction. Extraction methods involving EDTA, SDS, and proteinase-K resulted in low yield of genomic DNA. On the other hand, these methods resulted in high yield of genomic DNA from human cells suggesting the robust nature of A. castellanii DNases that are unaffected by reagents normally used in blocking eukaryotic DNases. In contrast, the use of chaotropic agent such as guanidine thiocyanate improved the yield of genomic DNA from A. castellanii cells significantly. Further purification and characterization of Acanthamoeba DNases is needed to study their non-classic distinct properties and to determine their role in the biology, cellular differentiation, cell cycle progression, and arrest of Acanthamoeba.

  14. Genotypic characterization of amoeba isolated from Acanthamoeba keratitis in Poland.

    Science.gov (United States)

    Derda, Monika; Solarczyk, Piotr; Cholewiński, Marcin; Hadaś, Edward

    2015-03-01

    Free-living amoebae belonging to the genus Acanthamoeba are the causative factor of many diseases. Among others, they cause Acanthamoeba keratitis (AK), a condition that usually occurs in contact lens wearers, though it is also observed in non-wearers. The number of diagnosed cases of AK increased more than eightfold during 8 years in the USA, and a proportional increase in frequency also occurred in Poland and Europe. Cases of AK are usually diagnosed late, and their therapy is difficult and rarely successful. AK is an uncommon diagnosis in Poland. The increased number of positive cases observed in our laboratory may reflect the growing at-risk population of contact lens wearers. Acanthamoeba as a genus of facultative human parasites is currently classified into 17 genotypes. Isolates belonging to seven genotypes were found to be associated with AK. One genotype in particular, T4, was found to be overrepresented in human disease. The main finding of our study is that in Poland, AK is almost always associated with the T4 genotype.

  15. Use of multiple immunosuppressive agents in recalcitrant ACANTHAMOEBA scleritis.

    Science.gov (United States)

    Igras, Estera; Murphy, Conor

    2015-04-15

    A 48-year-old woman who is a contact lens wearer presented with unilateral ACANTHAMOEBA keratitis, confirmed by PCR, which responded initially to topical polyhexamethylene biguanide (PHMB) and brolene. Three months later, despite continued treatment, she developed diffuse anterior scleritis with severe pain and marked scleral injection but without evidence of recurrence keratitis. Oral non-steroidal anti-inflammatories and oral high-dose corticosteroids were added without success. Subsequent treatment with intravenous methylprednisolone and high-dose cyclosporine led to a temporary improvement. Re-presenting with signs of recurrent scleritis and severe pain, the antitumor necrosis factor monoclonal antibody adalimumab, and later oral cyclophosphamide, were added. This led to complete quiescence of the scleritis. Unfortunately, frequent recurrences of ACANTHAMOEBA keratitis and anterior uveitis occurred on immunosuppression requiring continued treatment with PHMB, brolene and topical corticosteroids. This is the first case of severe refractory ACANTHAMOEBA scleritis requiring the concomitant use of four immunosuppressive agents to achieve continued disease control. The challenges in managing this case are discussed.

  16. The fine structure of the Acanthamoeba polyphaga cyst wall.

    Science.gov (United States)

    Lemgruber, Leandro; Lupetti, Pietro; De Souza, Wanderley; Vommaro, Rossiane C; da Rocha-Azevedo, Bruno

    2010-04-01

    Members of the genus Acanthamoeba are present in diverse environments, from freshwater to soil, and also in humans, causing serious brain and corneal infections. Their life cycle presents two stages: the dividing trophozoite and the quiescent cyst. The structures of these life stages have been studied for many years, and structural data have been used for taxonomy. The ultrastructural work on Acanthamoeba cysts was carried out previously by routine transmission electron microscopy (TEM), a process that requires the use of chemical fixation, a procedure that can cause serious artifacts in the ultrastructure of the studied material. In order to prevent fixation artifacts, we processed Acanthamoeba polyphaga cysts by ultrarapid freezing, followed by freeze-fracturing and deep-etching, in order to obtain a 3D visualization of the arrangements of the cyst wall. The exocyst presented an irregular surface, with vesicles located within or near this layer. The endocyst, instead, showed a biphasic arrangement with a more compact district in its innermost part, and a more loosened outer layer. For this reason, it was difficult to distinguish the filaments present in the intercyst space from those forming the endocyst. Surprisingly, the intercyst space was thinner when compared with samples processed by conventional TEM, evidencing the possible damage consequent to the use of chemical fixation.

  17. Balamuthia mandrillaris and Acanthamoeba Amebic Encephalitis with Neurotoxoplasmosis Coinfection in a Patient with Advanced HIV Infection

    OpenAIRE

    Pietrucha-Dilanchian, Paula; Chan, Joseph C.; Castellano-Sanchez, Amilcar; Hirzel, Alicia; Laowansiri, Panthipa; Tuda, Claudio; Visvesvara, Govinda S.; Qvarnstrom, Yvonne; Ratzan, Kenneth R.

    2012-01-01

    We describe a patient with advanced HIV infection and Balamuthia mandrillaris and Acanthamoeba amebic encephalitis with Toxoplasma gondii coinfection. A multidisciplinary effort and state-of-the-art diagnostic techniques were required for diagnosis. Our patient is the first reported case of an HIV-infected person with dual Balamuthia mandrillaris and Acanthamoeba amebic encephalitis with neurotoxoplasmosis coinfection.

  18. Microarray analysis of differentially expressed genes between cysts and trophozoites of Acanthamoeba castellanii.

    Science.gov (United States)

    Moon, Eun-Kyung; Xuan, Ying-Hua; Chung, Dong-Il; Hong, Yeonchul; Kong, Hyun-Hee

    2011-12-01

    Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

  19. Evidence for a previously unrecognized mycobacterial endosymbiont in Acanthamoeba castellanii strain Ma (ATCC ® 50370 ™).

    Science.gov (United States)

    Glaser, Kathleen C; Hetrick, Neil D; Molestina, Robert E

    2011-01-01

    We describe the isolation of a mycobacterium from Acanthamoeba castellanii strain Ma (ATCC(®) 50370(™)). The mycobacterium resides within vacuoles of A. castellanii, can be cultured by routine methodologies, and is a member of the Mycobacterium avium complex. Previously unrecognized mycobacterial endosymbionts are likely common among strains of Acanthamoeba housed at culture collections.

  20. Development of an immunochromatographic assay kit using fluorescent silica nanoparticles for rapid diagnosis of Acanthamoeba keratitis.

    Science.gov (United States)

    Toriyama, Koji; Suzuki, Takashi; Inoue, Tomoyuki; Eguchi, Hiroshi; Hoshi, Saichi; Inoue, Yoshitsugu; Aizawa, Hideki; Miyoshi, Kazutomi; Ohkubo, Michio; Hiwatashi, Eiji; Tachibana, Hiroshi; Ohashi, Yuichi

    2015-01-01

    We developed an immunochromatographic assay kit that uses fluorescent silica nanoparticles bound to anti-Acanthamoeba antibodies (fluorescent immunochromatographic assay [FICGA]) and evaluated its efficacy for the detection of Acanthamoeba and diagnosis of Acanthamoeba keratitis (AK). The sensitivity of the FICGA kit was evaluated using samples of Acanthamoeba trophozoites and cysts diluted to various concentrations. A conventional immunochromatographic assay kit with latex labels (LICGA) was also evaluated to determine its sensitivity in detecting Acanthamoeba trophozoites. To check for cross-reactivity, the FICGA was performed by using samples of other common causative pathogens of infectious keratitis, such as Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, and Candida albicans. Corneal scrapings from patients with suspected AK were tested with the FICGA kit to detect the presence of Acanthamoeba, and the results were compared with those of real-time PCR. The FICGA kit detected organisms at concentrations as low as 5 trophozoites or 40 cysts per sample. There were no cross-reactivities with other pathogens. The FICGA was approximately 20 times more sensitive than the LICGA for the detection of Acanthamoeba trophozoites. The FICGA kit yielded positive results for all 10 patients, which corresponded well with the real-time PCR results. The FICGA kit demonstrated high sensitivity for the detection of Acanthamoeba and may be useful for the diagnosis of AK.

  1. The Isolation and Detection of Acanthamoeba Keratitis in Rural Water Sources of Arak, Iran

    Directory of Open Access Journals (Sweden)

    Mosayebi, M. (PhD

    2014-05-01

    Full Text Available Background and Objective: Acanthamoeba species are free-living protozoa that can be isolated from all environments. They can bring about different diseases in healthy individuals and immune suppressed patients, for example, Granulomatous Amoebic Encephalitis (GAE, Acanthamoeba Keratitis (AK, Cutaneous and Nasopharyngeal infections. The aim of this study was to evaluate the Acanthamoeba prevalence in rural water sources of Markazi province. Material and Methods: In this cross sectional study, 54 water samples were collected from 36 villages of Markus province. First, the Samples were filtered by filter paper (watchman 42. Next, the filtered paper were placed in page saline solution and centrifuged. Then, the obtained sediment was cultured on non-nutrient agar (NNA plates overlaid with heat-killed Escherichia coli. After that, the provided smear (after 4 - 7 days stained with Geimsa. Results: The samples were positive (33; 61.11% and negative (21; 38.89% for Acanthamoeba cyst. The best result for isolation of Acanthamoeba cysts was obtained after shaking of filter paper. Conclusion: A high percentage of rural water sources have been contaminated with Acanthamoeba, which can be the major factor in causing human infections. Therefore, some effective methods are required to prevent from water sources contamination. Keywords: Acanthamoeba; Acanthamoeba Keratitis; Markazi Province; Rural; Water Sources

  2. Variabilidad genética de la respuesta inflamatoria: I. Polimorfismo -511 C/T en el gen IL1β en diferentes subpoblaciones peruanas

    Directory of Open Access Journals (Sweden)

    Óscar Acosta

    2012-07-01

    Full Text Available El polimorfismo -511 citosina/timina (-511 C/T en la región promotora del gen interleuquina 1 beta (IL1β estα implicado en la producciσn diferencial de la citoquina y por tanto puede estar asociado a la respuesta inmuno-inflamatoria en obesidad, dislipidemias, cardiopatías, cáncer, infecciones, y el tratamiento con nutrientes y fármacos. Objetivos: Establecer la distribución de frecuencias de los genotipos y alelos del polimorfismo -511 C/T del gen IL1β en diferentes subpoblaciones peruanas. Diseño: Estudio descriptivo, observacional, transversal. Instituciones: Centro de Investigación de Bioquímica y Nutrición e Instituto de Medicina Tropical D.A. Carrión, Facultad de Medicina, UNMSM y Centro de Genética y Biología Molecular, Facultad de Medicina, USMP, Lima, Perú. Participantes: Pobladores peruanos. Intervenciones: Extracción de ADN genómico a partir de muestras sanguíneas o epitelio bucal según metodología estándar, de 168 individuos de 9 grupos subpoblacionales: 23 mestizos de Lima, 33 amazónicos (20 de Pucallpa y 13 de Amazonas y 112 andinos (12 de Ancash, 10 de Cajamarca, 18 de Huarochirí-Lima, 25 de Puno-Taquile, 25 de Puno-Uros y 22 de Puno-Anapia. Análisis del polimorfismo -511 C/T mediante la técnica de PCR/RFLP, con primers específicos y digestión con la enzima de restricción AvaI, detectándose los fragmentos por electroforesis en geles de agarosa al 2% y tinción con bromuro de etidio. Principales medidas de resultados: Frecuencias genotípicas y alélicas del gen IL1β. Resultados: Se encontró las siguientes frecuencias genotípicas CC=0,024; CT=0,369 y TT=0,607, consistentes con el equilibrio de Hardy-Weinberg; y las frecuencias alélicas fueron alelo C=0,208 y aleloT= 0,792. La frecuencia del alelo T, considerado el mutante, fue muy alta en los Uros de Puno (0.940 y más baja en los mestizos de Lima (0.609. La comparación de las frecuencias genotípicas (TT versus CT+CC y alélicas (T versus C

  3. Efecto de un nuevo xanthanólido sesquiterpeno sobre la activación de mastocitos inducida por neurópeptidos pro-inflamatorios

    OpenAIRE

    Vargas, P.; Martino, E; Fogal, T.; Tonn, C.; Penissi, A.

    2009-01-01

    Los mastocitos son células del tejido conectivo que participan en la génesis y modulación de las respuestas inflamatorias celulares. En trabajos previos hemos demostrado que xanthatina (xanthanólido sesquiterpeno aislado de Xanthium cavanillesii Schouw) inhibe la activación de mastocitos inducida por secretagogos experimentales. Sin embargo, se desconoce su efecto sobre la activación de mastocitos inducida por estímulos fisiopatológicos. Estos...

  4. Characterisation and expression analysis of trophozoite and cyst proteins of Acanthamoeba spp. isolated from Acanthamoeba keratitis (AK) patient.

    Science.gov (United States)

    Behera, Himansu Sekhar; Satpathy, Gita

    2016-01-01

    The study was carried out to characterise and analyze the expression pattern of proteins of infective trophozoite and cyst forms of Acanthamoeba spp. isolated from an amoebic keratitis patient. Protein was isolated from the trophozoites and cysts of Acanthamoeba spp. isolates and subjected to SDS PAGE, 2D PAGE analysis where a large number of protein bands and protein spots were observed. Four prominent protein spots i.e. 2 from trophozoites and 2 from cysts that appeared more intense compared to the corresponding spots in other corresponding gel were excised from the 2D PAGE gels and analysed by MALDI-TOF/TOF MS assay and Mascot search software. Protein spots from trophozoites were identified as "hypothetical protein ACA1" and "eukaryotic porin protein" and those from cysts were identified as "chaperone protein DnaK" and "chaperonin protein" respectively. Proteomic results of 4 proteins were further validated by reverse genomics using quantitative real time PCR assay which showed a 1388 fold and 4.35 fold increase in expression of "hypothetical protein ACA1" gene and "eukaryotic porin protein" gene respectively in trophozoites compared to cysts and a 15 fold and 12.36 fold increase in expression of "chaperone protein DnaK" gene and "chaperonin protein" gene respectively in cysts compared to trophozoites. "Hypothetical protein ACA1" of trophozoites, whose function is unknown might have some important role in the parasite division and pathogenicty of Acanthamoeba spp. which needs further study. As trophozoites are the active and feeding form of Acanthamoeba spp., "eukaryotic porin" proteins may have some important role in efflux of toxic metabolites and exudates from interior of cell to outside along with some role in pathogenicity. Similarly proteins such as "chaperone protein DnaK" and "chaperonin protein" which belongs to group of heat shock proteins may have a role in folding of cyst specific proteins in cyst which needs further study.

  5. Voriconazole as a first-line treatment against potentially pathogenic Acanthamoeba strains from Peru.

    Science.gov (United States)

    Cabello-Vílchez, Alfonso Martín; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Reyes-Batlle, María; Sifaoui, Ines; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-02-01

    Pathogenic strains of Acanthamoeba genus are the causative agents of fatal granulomatous amoebic encephalitis and a serious sight-threatening infection of the eye known as Acanthamoeba keratitis. In a previous study, Acanthamoeba strains were isolated from nasal swabs collected from healthy individuals in Peru. In the present study, the pathogenic potential of the isolated strains was established based on temperature and osmotolerance assays as well as the secretion rate of extracellular proteases. Based on these experiments, four strains that showed the highest pathogenic potential were selected for sensitivity assays against two molecules (voriconazole and chlorhexidine) which are currently used for the treatment of Acanthamoeba infections. After performing sensitivity and activity assays, it was found that both drugs were active against the tested strains. However, voriconazole showed higher activity against the studied strains compared to chlorhexidine. Therefore, voriconazole should be established as a first-line treatment against Acanthamoeba infections at least in the studied region of Peru.

  6. Insights from the DNA databases: approaches to the phylogenetic structure of Acanthamoeba.

    Science.gov (United States)

    Fuerst, Paul A

    2014-11-01

    Species of Acanthamoeba have been traditionally described using morphology (primarily cyst structure), or cytology of nuclear division (used by Pussard and Pons, 1977). Twenty-plus putative species were proposed based on such criteria. Morphology, however, is often plastic, dependent upon culture conditions. DNA sequences of the nuclear small subunit (18S) rRNA that can be used for the study of the phylogeny of Acanthamoeba have increased from a single sequence in 1986 to more than 1800 in 2013. Some of the patterns of the sequence data for Acanthamoeba are reviewed, and some of the insights that this data illuminates are illustrated. In particular, the data suggest the existence of 20 or more genotypic types, a number not dissimilar to the number of named species of Acanthamoeba. However, molecular studies make clear that the relationship between phylogenetic relatedness and species names as we know them for Acanthamoeba is tenuous at best.

  7. Isolation and Molecular Characterization of Acanthamoeba Strains from Dental Units in Costa Rica.

    Science.gov (United States)

    Retana-Moreira, Lissette; Abrahams-Sandí, Elizabeth; Castro-Artavia, Esteban; Fernández-Sánchez, Ana; Castro-Castillo, Alfredo; Reyes-Batlle, María; Lorenzo-Morales, Jacob

    2015-01-01

    Free-living amoebae are protozoa widely distributed in nature, which can be found in a variety of environments. Four genera are recognized as causal agents of infections in humans and animals: Acanthamoeba, Naegleria, Balamuthia, and Sappinia. In this study, the presence of Acanthamoeba in dental units was determined and the isolates obtained were molecularly characterized; osmotolerance and thermotolerance assays were also performed to evaluate multiplication under these conditions, frequently associated with pathogenicity. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of Acanthamoeba genotype T4 in 14% of the units sampled. Osmotolerance and thermotolerance tests were positive for more than 80% of the isolates. Up to date, this is the first study that reports the detection, identification, and genotyping of Acanthamoeba isolated from dental units in Costa Rica and even in Latin-America. Further assays to determine the potential pathogenicity of these Acanthamoeba isolates are underway.

  8. Estudios clínicos de neumonitis por hipersensibilidad: Espartosis, suberosis y pulmón del cuidador de aves

    OpenAIRE

    Roger Reig, Àlex

    2013-01-01

    Las neumonitis por hipersensibilidad (NH) son un grupo de enfermedades producidas por la exposición a diferentes sustancias orgánicas, y por su consecuente inhalación, caracterizadas por la presencia de una reacción inflamatoria, en individuos susceptibles, que tiene lugar en alveolos, bronquiolos e intersticio pulmonar. En España, las NH ocupan el 5º lugar en el listado de enfermedades pulmonares intersticiales más frecuentemente diagnosticadas. El pulmón del cuidador de aves (PCA) es la ent...

  9. Importancia de los lípidos en el tratamiento nutricional de las patologías de base inflamatoria Importance of lipids in the nutritional treatment of inflammatory diseases

    Directory of Open Access Journals (Sweden)

    M. D. Mesa García

    2006-05-01

    Full Text Available En las últimas décadas, el conocimiento acerca de las propiedades antiinflamatorias de los lípidos ha facilitado el desarrollo de productos adaptados a la nutrición enteral y parenteral que han sido utilizados como terapia en numerosas enfermedades de base inflamatoria. En esta revisión se ofrece una visión general de los efectos de los lípidos utilizados en nutrición enteral, sobre ciertas patologías en cuya base están implicados procesos inflamatorios, como es el caso de las enfermedades inflamatorias intestinales, la aterosclerosis, la fibrosis pulmonar y la artritis reumatoide, entre otras. Durante los procesos inflamatorios se producen mediadores químicos, como los eicosanoides, a partir de ácidos grasos poliinsaturados presentes en las membranas celulares. La actividad inflamatoria de estas moléculas depende del ácido graso precursor. Los derivados del ácido araquidónico (de la serie n-6 tienen una elevada actividad proinflamatoria, mientras que los derivados del eicosapentaenoico (de la serie n-3 son débilmente inflamatorios. Así, al ingerir aceites de pescado, ricos en ácidos grasos n-3, aumenta el contenido de eicosapentaenoico y disminuye el araquidónico en las células del sistema inmune, por lo que se reduce la estimulación inflamatoria debido a la presencia de mediadores menos activos. Éste parece ser el mecanismo inmunomodulador fundamental de las dietas ricas en n-3. Asimismo, pero mediante mecanismos de acción diferentes, se ha demostrado que el ácido oleico disminuye la intensidad de algunos procesos inflamatorios, al disminuir la producción de mediadores quimiotácticos de inflamación. Podemos concluir que la suplementación de dietas para nutrición enteral con ácidos grasos poliinsaturados n-3 y oleico resulta beneficiosa para el tratamiento de ciertas patologías donde los procesos inflamatorios participan activamente en su desarrollo, mejorando su evolución y disminuyendo las dosis de f

  10. Análisis y aplicación de estrategias para modular la respuesta inflamatoria en patologías intestinales

    OpenAIRE

    Orsini Delgado, María Lucía

    2016-01-01

    Durante las últimas décadas se ha reportado un importante incremento en la incidencia de patologías inmunes inflamatorias, no infecciosas, fundamentalmente en países desarrollados o en vías de desarrollo. Debido a la velocidad con que se ha incrementado la incidencia de estas patologías, sólo los cambios en los factores ambientales son capaces de explicarlo. Estudios epidemiológicos han permitido observar una asociación entre el incremento en patologías inflamatorias no infecciosas y la dismi...

  11. Identifying differentially expressed genes in trophozoites and cysts of Acanthamoeba T4 genotype: Implications for developing new treatments for Acanthamoeba keratitis.

    Science.gov (United States)

    Abedkhojasteh, Hoda; Niyyati, Maryam; Rezaei, Sasan; Mohebali, Mehdi; Farnia, Shohreh; Kazemi-Rad, Elham; Roozafzoon, Reza; Sianati, Hamed; Rezaeian, Mostafa; Heidari, Mansour

    2015-02-01

    Acanthamoeba T4 genotype is the most prevalent genotype associated with amoebic keratitis. Acanthamoeba keratitis therapy is difficult due to transformation of trophozoite to cyst stage, which hinders the treatment of the disease. Although encystation assists the organism to survive against the chemotherapeutic compounds, the precise mechanism of encystation remains poorly understood. The purpose of this work was to identify differentially expressed genes in Acanthamoeba T4 genotype which might be useful for understanding of the encystment process and may thus help develop more efficient treatment. The mRNA profile of trophozoite and cyst of Acanthamoeba T4 genotype isolated from a soft contact lens wearer were analyzed using a cDNA amplified fragment length polymorphism (cDNA-AFLP) technique. Subsequently, a real time reverse transcriptase-PCR was performed to validate the cDNA-AFLP results. Three genes, heat shock protein70 (hsp70), actin-I and elongation factor-1alpha (EF-1α) were differentially expressed during Acanthamoeba differentiation. An in silico result predicted that transformation of trophozoite to cyst could be mediated through their cooperation with the protein partners interaction. Taken together, our experimental and bioinformatics findings suggested potential functions of hsp70, EF-1α and actin-I in differentiation of Acanthamoeba T4 genotype which may be useful in the design of an efficient therapeutic strategy in AK.

  12. Artemether Exhibits Amoebicidal Activity against Acanthamoeba castellanii through Inhibition of the Serine Biosynthesis Pathway.

    Science.gov (United States)

    Deng, Yihong; Ran, Wei; Man, Suqin; Li, Xueping; Gao, Hongjian; Tang, Wei; Tachibana, Hiroshi; Cheng, Xunjia

    2015-08-01

    Acanthamoeba sp. parasites are the causative agents of Acanthamoeba keratitis, fatal granulomatous amoebic encephalitis, and cutaneous infections. However, there are currently no effective drugs for these organisms. Here, we evaluated the activity of the antimalarial agent artemether against Acanthamoeba castellanii trophozoites and identified potential targets of this agent through a proteomic approach. Artemether exhibited in vitro amoebicidal activity in a time- and dose-dependent manner and induced ultrastructural modification and cell apoptosis. The iTRAQ quantitative proteomic analysis identified 707 proteins that were differentially expressed after artemether treatment. We focused on phosphoglycerate dehydrogenase and phosphoserine aminotransferase in the serine biosynthesis pathway because of their importance to the growth and proliferation of protozoan and cancer cells. The expression of these proteins in Acanthamoeba was validated using quantitative real-time PCR and Western blotting after artemether treatment. The changes in the expression levels of phosphoserine aminotransferase were consistent with those of phosphoglycerate dehydrogenase. Therefore, the downregulation of phosphoserine aminotransferase may be due to the downregulation of phosphoglycerate dehydrogenase. Furthermore, exogenous serine might antagonize the activity of artemether against Acanthamoeba trophozoites. These results indicate that the serine biosynthesis pathway is important to amoeba survival and that targeting these enzymes would improve the treatment of Acanthamoeba infections. Artemether may be used as a phosphoglycerate dehydrogenase inhibitor to control or block Acanthamoeba infections.

  13. A Case of Non-Contact Lens related Acanthamoeba keratitis in Malaysia

    Directory of Open Access Journals (Sweden)

    Mohamed Kamel, A. G.

    2005-01-01

    Full Text Available Acanthamoeba is a ubiquitous free-living amoeba and is responsible for an uncommon yet increasingly diagnosed keratitis in humans. Acanthamoeba keratitis is perhaps the most challenging ocular infection to manage successfully and it can result in permanently impaired vision or blindness. Although contact lens use is the principal risk factor, about 10% of cases occur following trauma and exposure to contaminated soil or water. Cases of Acanthamoeba keratitis involving contact lens wearers have previously been reported in Malaysia but this is the first time, a non-contact lens relatedAcanthamoeba keratitis is reported. The case involved a 28 year old Indonesian male construction worker who had a trauma of the right eye. While working his eye was struck by some sand and dust particles and he quickly washed his eye with water from an open tank at the construction site. He then experienced pain, redness, glaring and blurring of vision of the right eye. The diagnosis was missed at the initial presentation but subsequent culture of the corneal scraping demonstrated Acanthamoeba as the aetiological agent. The history, clinical findings, diagnosis and treatment of non-contact lens related Acanthamoeba keratitis are briefly discussed in this communication. We hope to create awareness especially among the medical and paramedical staff about the existence of this infection in the country and fully support the consideration of Acanthamoeba keratitis as part of the differential diagnosis of most cases of presumed microbial keratitis.

  14. Loop-mediated isothermal amplification targeting 18S ribosomal DNA for rapid detection of Acanthamoeba.

    Science.gov (United States)

    Yang, Hye-Won; Lee, Yu-Ran; Inoue, Noboru; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Kim, Hong-Kyun; Lee, Junhun; Goo, Youn-Kyoung; Kong, Hyun-Hee; Chung, Dong-Il; Hong, Yeonchul

    2013-06-01

    Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

  15. Abietane diterpenoids from Salvia sclarea transformed roots as growth inhibitors of pathogenic Acanthamoeba spp.

    Science.gov (United States)

    Kuźma, Łukasz; Derda, Monika; Hadaś, Edward; Wysokińska, Halina

    2015-01-01

    Amoebae from the genus Acanthamoeba are known agents leading to various diseases such as granulomatous amoebic encephalitis (GAE), a chronic progressive disease of the central nervous system, amoebic keratitis (AK), chronic eye infection, amoebic pneumitis (AP), chronic lung infection, and skin infections. It is known that various synthetic anti-Acanthamoeba substances are ineffective. Therefore, other substances, e.g., natural plant compounds, are the focus of biological investigations regarding anti-parasite activity. In this work, the ability of four abietane diterpenoids (ferruginol, salvipisone, aethiopinone, and 1-oxo-aethiopinone) to inhibit Acanthamoeba growth is reported. All investigated compounds were active against Acanthamoeba growing in vitro. Among them, ferruginol demonstrated the highest activity against Acanthamoeba. This compound inhibited Acanthamoeba growth by about 72% in a 3-day exposure period (IC50 17.45 μM), while aethiopinone and 1-oxo-aethiopinone demonstrated this activity at the level of 55-56%. Salvipisone reduced the growth of Acanthamoeba in vitro culture by 39%. For this compound, the value of IC50 was 701.94 μM after 72 h of exposure.

  16. Isolation and genotyping of Acanthamoeba spp. from recreational soil of parks in Tehran, Iran

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    M Niyyati

    2013-11-01

    Full Text Available Background & aim: Acanthamoeba is a genus of free-living amoebae found in environmental sources. These amphizoic amoebae can lead to severe human disease such as encephalitis and keratitis. Acanthamoeba transmits to humans through contact with soil and dust from scratching the skin. The aim of the present study was to identify the genotypes of Acanthamoeba in parks of the city of Tehran using molecular and morphological - based methods. Methods: In this study, 52 samples of soil were collected from 17 parks in Tehran. Samples were then filtered and cultured on 1.5% non-nutrient agar. DNA extraction and PCR amplification was performed using genus specific primers. Sequencing analysis and BLAST search were done for genotype identification. Results: Out of 52 soil samples, 14 strain (26.9% were positive for Acanthamoeba amoebae by microscopic observation. Out Of 14 positive isolates, 9 (17.3% were positive for Acanthamoeba using genus specific primer pairs. Of 14 strains, 9 were sequenced successfully. Genotype identification was revealed that all strains were belonged to T4 type. T4 genotypes among strains are human pathogens. Conclusions: Identification of pathogenic Acanthamoeba belonging to T4 genotype in recreational parks could be of utmost importance. Results of this study show that soil contamination, particularly in parks where children play and assemble is a sanitary risk for them. Key words: Acanthamoeba, Genotypes, Soil, Park

  17. A method for microbial decontamination of Acanthamoeba cultures using the peritoneal cavity of mice简

    Institute of Scientific and Technical Information of China (English)

    Daniella; de; Sousa; Mendes; Moreira; Alves; Rodrigo; Gurgel-Gon?alves; Patrícia; Albuquerque; César; Augusto; Cuba-Cuba; Maria; Imaculada; Muniz-Junqueira; Selma; Aparecida; Souza; Kückelhaus

    2015-01-01

    Objective: To evaluate whether the inoculation of contaminated cultures in the peritoneal cavity of mice could implement decontamination of Acanthamoeba cultures.Methods: Suspensions of Acanthamoeba, Acanthamoeba polyphaga ATCC 30461, or Acanthamoeba spp. isolated from soil(Un B13 strain) were inoculated in the peritoneal cavity of Swiss mice(n = 24). After 1, 6, 12, or 24 h of exposure the peritoneal cavity was washed and assessed for the presence of bacteria, fungi, and Acanthamoeba.Results: After 1 h of intraperitoneal inoculation at least 97% of the bacteria and 96% of the fungi(P < 0.05) and 99% of the bacteria(P < 0.05) were successfully eliminated from the ATCC 30461 strain and from the soil isolate Un B13 strain, respectively. This method also allowed the recovery of most trophozoites and cysts from both Acanthamoeba cultures at the end of 24 h.Conclusions: Our data demonstrated that this technique has great potential for decontamination of Acanthamoeba cultures in a short period of time.

  18. Mechanisms associated with phagocytosis of Arcobacter butzleri by Acanthamoeba castellanii.

    Science.gov (United States)

    Medina, Gustavo; Flores-Martin, Sandra; Fonseca, Belchiolina; Otth, Carola; Fernandez, Heriberto

    2014-05-01

    Acanthamoeba castellanii is a free-living amoeba widely found in environmental matrices such as soil and water. Arcobacter butzleri is an emerging potential zoonotic pathogen that can be isolated from environmental water sources, where they can establish endosymbiotic relationships with amoebas. The aim of this study was to describe the implication of mannose-binding proteins and membrane-associated receptors of glucose and galactose present in the amoebic membrane, during the attachment of Arcobacter butzleri by blocking with different saccharides. Another objective was to describe the signaling pathways involved in phagocytosis of these bacteria using specific inhibitors and analyze the implication of phagolysosome formation on the survival of Arcobacter butzleri inside the amoeba. We infer that the attachment of Arcobacter butzleri to the amoeba is a process which involves the participation of mannose-binding proteins and membrane-associated receptors of glucose and galactose present in the amoeba. We also demonstrated an active role of protozoan actin polymerization in the phagocytosis of Arcobacter butzleri and a critical involvement of PI3K and RhoA pathways. Further, we demonstrated that the tyrosine kinase-induced actin polymerization signal is essential in Acanthamoeba-mediated bacterial uptake. Through phagolysosomal formation analysis, we conclude that the survival of Arcobacter butzleri inside the amoeba could be related with the ability to remain inside vacuoles not fused with lysosomes, or with the ability to retard the fusion between these structures. All these results help the understanding of the bacterial uptake mechanisms used by Acanthamoeba castellanii and contribute to evidence of the survival mechanisms of Arcobacter butzleri.

  19. Acanthamoeba spp. in Contact Lenses from Healthy Individuals from Madrid, Spain

    Science.gov (United States)

    Gomes, Thiago dos Santos; Magnet, Angela; Izquierdo, Fernando; Vaccaro, Lucianna; Redondo, Fernando; Bueno, Sara; Sánchez, Maria Luisa; Angulo, Santiago; Fenoy, Soledad; Hurtado, Carolina; del Aguila, Carmen

    2016-01-01

    Purpose Acanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus aureus. Methods One hundred and seventy-seven healthy individuals from Madrid contributed their discarded CL and answered a questionnaire on hygiene habits. DNA was extracted from the CL solution and analyzed by real-time PCR for Acanthamoeba, Pseudomonas aeruginosa and Staphylococcus aureus. These CL and their solutions were also cultured on non-nutrient agar to isolate Acanthamoeba. Results Among the 177 samples, Acanthamoeba DNA was detected in 87 (49.2%), P. aeruginosa DNA in 14 (7.9%) and S. aureus DNA in 19 (10.7%). Cultivable amoebae, however, were observed in only one sample (0.6%). This isolate was genotyped as T4. The habits reported by this CL owner included some recognized risk factors for AK, but in this study only the practice of “not cleaning the CL case” presented some statistical significant association with Acanthamoeba DNA presence. Detection of the investigated bacterial DNA did not demonstrate statistical significant association with the studied practices, but the presence of P. aeruginosa revealed a possible inhibition of Acanthamoeba in these samples. Conclusions The PCR results suggest a high presence of Acanthamoeba spp. in healthy CL wearers from Madrid, but we can assume that CL solutions are properly disinfecting the CL since only 1.1% of the positive PCR samples correspond to viable amoebae and, after four years, only one participant reported stronger ocular problems. Nevertheless, more studies are necessary to corroborate this hypothesis. PMID

  20. Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis

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    Sharma Savitri

    2001-01-01

    Full Text Available Purpose: We describe a simple procedure of Immunoperoxidase (IP technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. Methods: A high titre polyclonal antibody against a local clinical isolate (axenic of Acanthamoeba species (trophozoite lysate antigen was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1 and patients showing no cysts in similar scrapings (pool-2, were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison with calcofluor white staining and culture. Results: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%. Conclusion: The immunoperoxidase technique is a simple and useful test in the diagnosis of

  1. Impacto de la nutrición en la evolución de la enfermedad inflamatoria intestinal Impact of nutritional treatment in the evolution of inflammatory bowel disease

    Directory of Open Access Journals (Sweden)

    M.ª D. Ballesteros Pomar

    2010-04-01

    Full Text Available La enfermedad inflamatoria intestinal es una entidad de etiopatogenia aún no bien conocida, con importantes implicaciones nutricionales y metabólicas, por la alta prevalencia de malnutrición que conlleva; por la posible implicación de factores dietéticos en su patogenia; y por la hipótesis de que la intervención nutricional pudiera ser un tratamiento primario de la enfermedad. Algunos nutrientes, además de su función exclusivamente nutricional, podrían inducir un bajo estímulo antigénico, regular respuestas inflamatorias e inmunológicas y estimular el trofismo de la mucosa intestinal. La evidencia disponible actual apoya el empleo de nutrición enteral en enfermedad de Crohn como terapia primaria en adultos si el tratamiento con corticoides no es posible (fracaso o contraindicación (grado de recomendación A o bien en terapia combinada con fármacos en pacientes malnutridos y estenosis inflamatoria del intestino. En los pacientes en remisión clínica duradera no se ha demostrado beneficio de la nutrición enteral o suplementos en ausencia de déficits nutricionales. No se recomienda el uso de fórmulas elementales ni modificadas (glutamina, ácidos grasos omega 3. En colitis ulcerosa, no se ha demostrado la influencia de la nutrición sobre la actividad de la enfermedad, aunque disponemos de datos prometedores sobre el papel de los ácidos grasos w3 con cubierta entérica y de un posible papel de los probióticos. En el tratamiento y profilaxis de la pouchitis crónica, el empleo de probióticos puede tener un papel (VSL#3. La nutrición debe considerarse un componente integral en el manejo de los pacientes con EII.Inflammatory bowel disease is an entity with not wellknown pathogenesis, and important nutritional and metabolic implications because of the high prevalence of malnutrition, the possible implication of dietary factors in its pathogenesis and because of the hypothesis that nutritional intervention could be a primary

  2. Protozoan Acanthamoeba polyphaga as a potential reservoir for Campylobacter jejuni.

    Science.gov (United States)

    Axelsson-Olsson, Diana; Waldenström, Jonas; Broman, Tina; Olsen, Björn; Holmberg, Martin

    2005-02-01

    We showed by a laboratory experiment that four different Campylobacter jejuni strains are able to infect the protozoan Acanthamoeba polyphaga. C. jejuni cells survived for longer periods when cocultured with amoebae than when grown in culture alone. The infecting C. jejuni cells aggregated in amoebic vacuoles, in which they were seen to be actively moving. Furthermore, a resuscitation of bacterial cultures that were previously negative in culturability tests was observed after reinoculation into fresh amoeba cultures. After spontaneous rupture of the amoebae, C. jejuni could be detected by microscopy and culturability tests. Our results indicate that amoebae may serve as a nonvertebrate reservoir for C. jejuni in the environment.

  3. Acil Coenzima-A sintetasas de cadena larga (ACSLs) como biomarcadores cito-serológicos en enfermedades inflamatorias o autoinmunes

    OpenAIRE

    Matesanz, F.; Fedetz, María; Ndagire, D.; Alcina, Antonio; Sabio, José Mario

    2010-01-01

    Uso de las Acil-Coenzima-A sintetasas de cadena larga (ACSL) como biomarcadores cito-serológicos de enfermedades inflamatorias o autoinmunes, y especialmente de lupus eritematoso sistémico, y método de obtención de datos útiles para el diagnóstico y seguimiento de dichas enfermedades. Venta

  4. Utilidad de la determinación sérica de microRNA-21 en la enfermedad inflamatoria del colon

    Directory of Open Access Journals (Sweden)

    P. Menéndez

    2013-04-01

    Conclusiones: Es necesario continuar la búsqueda de cuáles son los microRNA más apropiados para que su determinación en suero conlleve una mayor precisión en el establecimiento del diagnóstico y pronóstico de las enfermedades inflamatorias intestinales.

  5. Enhanced killing of Acanthamoeba cysts with a plant peroxidase-hydrogen peroxide-halide antimicrobial system.

    Science.gov (United States)

    Hughes, Reanne; Andrew, Peter W; Kilvington, Simon

    2003-05-01

    The activity of H(2)O(2) against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H(2)O(2), and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P Acanthamoeba cysts and prevention of acanthamoeba keratitis.

  6. Disseminated Acanthamoeba sinusitis in a patient with AIDS: a possible role for early antiretroviral therapy.

    Science.gov (United States)

    Carter, Wendy W; Gompf, Sandra G; Toney, John F; Greene, John N; Cutolo, Edward P

    2004-01-01

    Acanthamoeba, a free-living ameba, has been reported to infect humans with subacute encephalitis, sinusitis, or keratitis. Multiple cases of Acanthamoeba sinusitis with dissemination have been reported in association with AIDS, with high mortality. We report successful treatment of a 35-year-old woman who presented with sinusitis that progressed to disseminated acanthamebiasis as her initial manifestation of AIDS. To our knowledge, our patient was one of the few and longest-lived survivors of disseminated Acanthamoeba infection with AIDS. As with other opportunistic infections, early aggressive therapy including HAART may alter the outcome in this almost uniformly fatal disease.

  7. Pathogenic and nonpathogenic Acanthamoeba spp. in thermally polluted discharges and surface waters

    Energy Technology Data Exchange (ETDEWEB)

    de Jonckheere, J.F.

    1981-02-01

    During spring and autumn, the total number of amoebae and the number of acanthamoeba species able to grow at 37 degrees C were determined in six thermally polluted factory discharges and the surrounding surface waters. The isolated Acanthamoeba strains were studied for growth in axenic medium, cytopathic effect in Vito cell cultures, and virulence in mice. Although more amoebae were isolated in autumn, the number of Acanthamoeba species was lower than in spring, when the percent of pathogenic strains among the isolates was highest. Higher concentrations of amoebae were found in warm discharges, and more virulent strains occurred in thermal discharges than in surface waters.

  8. Acanthamoeba differentiation: a two-faced drama of Dr Jekyll and Mr Hyde.

    Science.gov (United States)

    Siddiqui, Ruqaiyyah; Dudley, Ricky; Khan, Naveed Ahmed

    2012-06-01

    The ability of cyst-forming protists such as Acanthamoeba to escape death by transforming into a cyst form, that is resistant to harsh physiological, environmental and pharmacological conditions, has continued to pose a serious challenge to human and animal health. A complete understanding of the fundamental principles of genome evolution and biochemical pathways of cellular differentiation offers unprecedented opportunities to counter detrimental outcomes. Acanthamoeba can elude inhospitable conditions by forming cysts. Here we unravel the processes involved in the phenotypic switching of Acanthamoeba, which are critical in our efforts to find potential targets for chemotherapy.

  9. Nuevas consideraciones fisiopatológicas sobre el síndrome de respuesta inflamatoria sistémica relacionada con la sepsis

    Directory of Open Access Journals (Sweden)

    Vivian R. Mena Miranda

    1996-04-01

    Full Text Available El shock séptico constituye una de las primeras causas de morbilidad y mortalidad en las unidades de cuidados intensivos y forma parte de los estadios evolutivos la respuesta inflamatoria sistémica (SRIS en relación con la infección, que fueron modificados por Jaffari para su aplicación en la edad pediátrica. Está relacionado con la exposición del organismo a agentes exógenos y/o mediadores endógenos, que producen una inadecuada perfusión hística con alteración en el consumo de oxígeno, en la permeabilidad vascular y la aparición de trastornos hemodinámicos que comprometen la vida del paciente. Se considera que el mediador central en el shock séptico es el factor de necrosis tumoral-a(FNT- a, aunque también responden de forma descontrolada otras citocinas, los polimorfonucleares, el sistema de contacto, el fibrinolítico, el de la coagulación, así como otras sustancias endógenas. El conocimiento de su fisiopatología hace posible el diagnóstico y tratamiento precoz, sobre todo en los grupos de riesgo, y la reducción de la mortalidad en las unidades de terapia intensiva por esta afección.The septic shock is one of the first causes of morbidity and mortality in intensive care units, and the systemic inflammatory response syndrome (SIRS is a part of the evolutive stages related to the infection, that were modified by Jaffari for its application in the pediatric age. Septic shock is related with the organism exposition to exogenous agents, and/or endogenous mediators that produce an inadequate histic perfusion with alteration in oxygen consumption, in vascular permeability, and the presentation of hemodynamic disorders that compromise the patient's life. It is considered that the main mediator in septic shock is the tumoral necrosis factor - (TBF- , although other cytosines also reply in an uncontrolled fashion, the polymorphonuclears, the contact system, the fibrinolytic, the coagulation one, as well as other endogenous

  10. Citocinas anti-inflamatorias y sus acciones y efectos en la sepsis y el choque séptico - Anti-inflammatory cytokines and their actions and effects in the sepsis and septic shock

    OpenAIRE

    González Álvarez, Ricardo; Zamora Rodríguez, Zullit; Alonso González, Yaima

    2009-01-01

    ResumenLas citocinas anti- inflamatorias reconocidas son las interleucinas (IL) IL-4, IL-10, IL-6, IL-13, el factor estimulante de colonia granulocito –macrófago (FEC-GM) y el interferón alfa (IFN-alfa), los cuales tienen la capacidad de inhibir la liberación de las citocinas pro-inflamatorias y de inducir la producción del antagonista del receptor de la IL-1 y la liberación del receptor soluble del FNT, los cuales limitan algunas de las actividades de las citocinas pro-inflamatorias IL-1 y F...

  11. New long chain bases in lipophosphonoglycan of Acanthamoeba castellanii.

    Science.gov (United States)

    Karaś, Magdalena A; Russa, Ryszard

    2013-06-01

    The polymer called lipophosphonoglycan (LPG) was isolated from Acanthamoeba castellanii membranes after exhaustive delipidation and butanol extraction. A novel extremely long phytosphingosine was revealed in glycoinositolphosphosphingolipid (GIPSL). All data obtained by gas-liquid chromatography coupled with MS analyses of products liberated during acid methanolysis and products of sodium metaperiodate and permanganate-periodate oxidations showed an unusual pattern of long chain bases (LCB) with branched bases (anteiso-C₂₄, anteiso-C₂₅, anteiso-C₂₆, iso-C₂₆, anteiso-C₂₇, and anteiso-C28) and normal ones (C₂₄, C₂₅, C₂₆, C₂₇). The phytosphingosines with hexa-, hepta-, and octacosanoic chains have not been detected in Acanthamoeba cells up to now. Also, the isomer configuration of long chain bases in LPG of A. castellanii was not defined in earlier reports. In the GC-MS chromatograms, the component forming a peak corresponding to anteiso-C₂₅ phytosphingosine was the most abundant and constituted more than 50 % of all LCB.

  12. Effects of mannose on pathogenesis of Acanthamoeba castellanii.

    Science.gov (United States)

    Yoo, Kyung-Tae; Jung, Suk-Yul

    2012-12-01

    Acanthamoeba spp. are single-celled protozoan organisms that are widely distributed in the environment. In this study, to understand functional roles of a mannose-binding protein (MBP), Acanthamoeba castellanii was treated with methyl-alpha-D-mannopyranoside (mannose), and adhesion and cytotoxicity of the amoeba were analyzed. In addition, to understand the association of MBP for amoeba phagocytosis, phagocytosis assay was analyzed using non-pathogenic bacterium, Escherichia coli K12. Amoebae treated with mannose for 20 cycles exhibited larger vacuoles occupying the most area of the amoebic cytoplasm in comparison with the control group amoebae and glucose-treated amoebae. Mannose-selected amoebae exhibited lower levels of binding to Chinese hamster ovary (CHO) cells. Exogenous mannose inhibited >50% inhibition of amoebae (control group) binding to CHO cells. Moreover, exogenous mannose inhibited amoebae (i.e., man-treated) binding to CHO cells by <15%. Mannose-selected amoebae exhibited significantly decreased cytotoxicity to CHO cells compared with the control group amoebae, 25.1% vs 92.1%. In phagocytic assay, mannose-selected amoebae exhibited significant decreases in bacterial uptake in comparison with the control group, 0.019% vs 0.03% (P<0.05). Taken together, it is suggested that mannose-selected A. castellanii trophozoites should be severely damaged and do not well interact with a target cell via a lectin of MBP.

  13. Encystation in Acanthamoeba castellanii: development of biocide resistance.

    Science.gov (United States)

    Lloyd, D; Turner, N A; Khunkitti, W; Hann, A C; Furr, J R; Russell, A D

    2001-01-01

    Since the early 1960s, axenic culture and the development of procedures for the induction of encystation have made Acanthamoeba spp. superb experimental systems for studies of cell biology and differentiation. More recently, since their roles as human pathogens causing keratitis and encephalitis have become widely recognized, it has become urgent to understand the parameters that determine differentiation, as cysts are much more resistant to biocides than are the trophozoites. Viability of trophozoites of the soil amoeba Acanthamoeba castellanii (Neff), is conveniently measured by its ability to form plaques on a lawn of Escherichia coli. Use of confocal laser scanning microscopy with Calcofluor white, Congo Red or the anionic oxonol dye, DiBAC4(3) or flow cytometry with propidium iodide diacetate and fluorescein or oxonol provides more rapid assessment. For cysts, the plaque method is still the best, because dye exclusion does not necessarily indicate viability and therefore the plate count method has been used to study the sequence of development of biocide resistance during the differentiation process. After two hours, resistance to HCl was apparent. Polyhexamethylene biguanide, benzalkonium chloride, propamidine isethionate, pentamidine isethionate, dibromopropamine isethionate, and H2O2 and moist heat, all lost effectiveness at between 14 and 24 h after trophozoites were inoculated into encystation media. Chlorhexidine diacetate resistance was observed at between 24 and 36 h. The molecular biology and biochemistry of the modifications that underlie these changes are now being investigated.

  14. Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga.

    Science.gov (United States)

    Caumo, Karin Silva; Monteiro, Karina Mariante; Ott, Thiely Rodrigues; Maschio, Vinicius José; Wagner, Glauber; Ferreira, Henrique Bunselmeyer; Rott, Marilise Brittes

    2014-12-01

    Acanthamoeba polyphaga is a free-living protozoan pathogen, whose infective trophozoite form is capable of causing a blinding keratitis and fatal granulomatous encephalitis in humans. The damage caused by A. polyphaga trophozoites in human corneal or brain infections is the result of several different pathogenic mechanisms that have not yet been elucidated at the molecular level. We performed a comprehensive analysis of the proteins expressed by A. polyphaga trophozoites, based on complementary 2-DE MS/MS and gel-free LC-MS/MS approaches. Overall, 202 non-redundant proteins were identified. An A. polyphaga proteomic map in the pH range 3-10 was produced, with protein identification for 184 of 370 resolved spots, corresponding to 142 proteins. Additionally, 94 proteins were identified by gel-free LC-MS/MS. Functional classification revealed several proteins with potential importance for pathogen survival and infection of mammalian hosts, including surface proteins and proteins related to defense mechanisms. Our study provided the first comprehensive proteomic survey of the trophozoite infective stage of an Acanthamoeba species, and established foundations for prospective, comparative and functional studies of proteins involved in mechanisms of survival, development, and pathogenicity in A. polyphaga and other pathogenic amoebae.

  15. Acanthamoeba keratitis: study of the 5-year incidence in Israel.

    Science.gov (United States)

    Graffi, Shmuel; Peretz, Avi; Jabaly, Haneen; Koiefman, Anna; Naftali, Modi

    2013-11-01

    Acanthamoeba keratitis (AK) is not a notifiable disease in Israel, so there are no accurate incidence rates for this condition in Israel. The aim of this study was to estimate the incidence of AK in Israel for the years 2008-2012. We distributed a survey questionnaire to laboratory managers in Israel. The laboratories were affiliated to medical institutes that either provided ophthalmology services or served community ophthalmology clinics. Our questionnaire requested survey respondents to provide information regarding the methods used to diagnose AK, and the number of positive and negative cultures for Acanthamoebae species performed for each of the years from 2008 to 2012. Six laboratories used non-nutrient agar with Escherichia coli as the culture medium, one used calcofluor-white staining with fluorescent microscopy, and two used PCR for diagnosing AK. Twenty-three AK cases were identified, to give an estimated incidence of 1/1 668 552. AK is mostly attributable to the use of contact lenses. As contact lenses are popular in Israel, we expected a higher incidence rate. A lower than expected incidence rate may indicate insufficient awareness of AK in Israel.

  16. 18S Ribosomal DNA Typing and Tracking of Acanthamoeba Species Isolates from Corneal Scrape Specimens, Contact Lenses, Lens Cases, and Home Water Supplies of Acanthamoeba Keratitis Patients in Hong Kong

    OpenAIRE

    Booton, G. C.; Kelly, D J; Chu, Y.-W.; Seal, D. V.; Houang, E.; Lam, D S C; Byers, T. J.; Fuerst, P A

    2002-01-01

    We examined partial 18S ribosomal DNA (Rns) sequences of Acanthamoeba isolates cultured in a study of microbial keratitis in Hong Kong. Sequence differences were sufficient to distinguish closely related strains and were used to examine links between strains obtained from corneal scrape specimens, contact lenses, lens cases, lens case solutions, and home water-supply faucets of patients with Acanthamoeba. We also looked for evidence of mixed infections. Identification of Acanthamoeba Rns geno...

  17. Azole Antifungal Agents To Treat the Human Pathogens Acanthamoeba castellanii and Acanthamoeba polyphaga through Inhibition of Sterol 14α-Demethylase (CYP51).

    Science.gov (United States)

    Lamb, David C; Warrilow, Andrew G S; Rolley, Nicola J; Parker, Josie E; Nes, W David; Smith, Stephen N; Kelly, Diane E; Kelly, Steven L

    2015-08-01

    In this study, we investigate the amebicidal activities of the pharmaceutical triazole CYP51 inhibitors fluconazole, itraconazole, and voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against Acanthamoeba infections in humans. Amebicidal activities of the triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition, triazole effectiveness was assessed by ligand binding studies and inhibition of CYP51 activity of purified A. castellanii CYP51 (AcCYP51) that was heterologously expressed in Escherichia coli. Itraconazole and voriconazole bound tightly to AcCYP51 (dissociation constant [Kd] of 10 and 13 nM), whereas fluconazole bound weakly (Kd of 2,137 nM). Both itraconazole and voriconazole were confirmed to be strong inhibitors of AcCYP51 activity (50% inhibitory concentrations [IC50] of 0.23 and 0.39 μM), whereas inhibition by fluconazole was weak (IC50, 30 μM). However, itraconazole was 8- to 16-fold less effective (MIC, 16 mg/liter) at inhibiting A. polyphaga and A. castellanii cell proliferation than voriconazole (MIC, 1 to 2 mg/liter), while fluconazole did not inhibit Acanthamoeba cell division (MIC, >64 mg/liter) in vitro. Voriconazole was an effective inhibitor of trophozoite proliferation for A. castellanii and A. polyphaga; therefore, it should be evaluated in trials versus itraconazole for controlling Acanthamoeba infections.

  18. Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus.

    Science.gov (United States)

    Hansen, Birgitte; Kronborg, Gitte

    2003-01-01

    Acanthamoeba keratitis is potentially blinding and often associated with contact lens wearing. A human immunodeficiency virus (HIV)-positive patient, a non-contact lens wearer, presented with keratitis. She experienced a protracted course of disease, characterized by exacerbations and remissions, and was treated with various topical antibiotics and steroids. 13 months after symptom onset the eye was removed owing to serious scarring of cornea and unbearable pain. Microbiological and histopathological examination of the cornea showed Acanthamoeba. In non-contact lens wearers suffering from Acanthamoeba keratitis the diagnosis is delayed, pathognomonic features are often not seen and visual outcome is usually poor. There is no known relation between HIV infection and Acanthamoeba keratitis.

  19. Diagnosis of acanthamoeba keratitis-A report of four cases and review of literature

    Directory of Open Access Journals (Sweden)

    Sharma Savitri

    1990-01-01

    Full Text Available Acanthamoeba keratitis is not reported often in India. We reported the first case diagnosed in this country a year back. In this communication, four more cases of Acanthamoeba keratitis diagnosed since then are being reported along with a brief review of literature. Diagnosis in all these patients was based on observation of acanthamoeba cysts in 10% KOH wet mount of corneal ulcer scrapings and subsequent culture. The characteristic ring infiltration of cornea was seen in all of them. All cases were treated medically with the available antiamoebic drugs (Miconazole, Neosporin, Ketoconazole and Metronidazole in different combinations. Only one out of four, responded with complete healing of the ulcer. Acanthamoeba keratitis is probably not as uncommon in India as it is thought to be. With increased awareness and performance of minimal laboratory tests the condition may be diagnosed more often.

  20. Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus

    DEFF Research Database (Denmark)

    Hansen, Birgitte Rønde; Kronborg, Gitte

    2003-01-01

    Acanthamoeba keratitis is potentially blinding and often associated with contact lens wearing. A human immunodeficiency virus (HIV)-positive patient, a non-contact lens wearer, presented with keratitis. She experienced a protracted course of disease, characterized by exacerbations and remissions......, and was treated with various topical antibiotics and steroids. 13 months after symptom onset the eye was removed owing to serious scarring of cornea and unbearable pain. Microbiological and histopathological examination of the cornea showed Acanthamoeba. In non-contact lens wearers suffering from Acanthamoeba...... keratitis the diagnosis is delayed, pathognomonic features are often not seen and visual outcome is usually poor. There is no known relation between HIV infection and Acanthamoeba keratitis....

  1. Acanthamoeba and Stenotrophomonas maltophilia keratitis with fungal keratitis in the contralateral eye

    Directory of Open Access Journals (Sweden)

    Thomas F Mauger

    2010-10-01

    Full Text Available Thomas F Mauger, Rebecca Ann Kuennen, Reynell Harder Smith, William SawyerDepartment of Ophthalmology, The Ohio State University, Columbus, OH, USAPurpose: The purpose of this study is to describe the diagnosis, course, and outcome of a case of Acanthamoeba and Stenotrophomonas keratitis with a fungal keratitis in the contralateral eye.Methods: A case of Acanthamoeba and Stenotrophomonas keratitis was diagnosed with confocal microscopy and cultures with confocal diagnosis of fungal keratitis in the fellow eye.Results: During the initial treatment of the Acanthamoeba and Stenotrophomonas keratitis, the contralateral eye developed a keratitis that demonstrated hyphae in the corneal stroma with confocal microscopy consistent with fungal keratitis.Conclusions: Bilateral chronic keratitis cannot be assumed to be caused by the same organism and independent cultures, and confocal microscopy needs to be performed to direct appropriate therapy.Keywords: Acanthamoeba, Stenotrophomonas, confocal, fungus, keratitis

  2. Evaluation of Acanthamoeba isolates from environmental sources in Tenerife, Canary Islands, Spain.

    Science.gov (United States)

    Lorenzo-Morales, Jacob; Monteverde-Miranda, Carlos A; Jiménez, Concepción; Tejedor, María Luisa; Valladares, Basilio; Ortega-Rivas, Antonio

    2005-01-01

    Tests for potentially pathogenic amoebae were carried out in order to determine the presence of free-living amoebae of genus Acanthamoeba in soil and beach sand sources related to human environments in Tenerife, Canary Islands, Spain. Acanthamoeba identification was based on the morphology of cyst and trophozoite forms and PCR amplification with a genus specific primer pair. The pathogenical potential of Acanthamoeba isolates was characterized by temperature and osmotolerance assays and PCR reactions with two primer pairs related to Acanthamoeba pathogenesis. The results demonstrate the presence of potential pathogenic strain in both sources. Thus, some of the amoebae found in these habitats could act as opportunistic pathogens and may present a risk to human health.

  3. Isolation and identification of pathogenic Acanthamoeba strains in Tenerife, Canary Islands, Spain from water sources.

    Science.gov (United States)

    Lorenzo-Morales, Jacob; Ortega-Rivas, Antonio; Foronda, Pilar; Martínez, Enrique; Valladares, Basilio

    2005-03-01

    A comprehensive survey to document the presence of free-living amoebae of the genus Acanthamoeba was conducted in tap water and sea water sources related to human environments in Tenerife, Canary Islands, Spain. Acanthamoeba identification was based on the morphology of cyst and trophozoite forms and PCR amplification with a genus-specific primer pair. The pathogenic potential of Acanthamoeba isolates was characterized by temperature and osmotolerance assays and PCR reactions with two primer pairs related to Acanthamoeba pathogenesis. The results demonstrate the presence of potentially pathogenic strains in both sources. Thus, some of the amoebae in these aquatic habitats can act as opportunistic pathogens, could play a role in the diseases of aquatic organisms, and may present a risk to human health.

  4. Juglone induces cell death of Acanthamoeba through increased production of reactive oxygen species.

    Science.gov (United States)

    Jha, Bijay Kumar; Jung, Hui-Jung; Seo, Incheol; Suh, Seong-Il; Suh, Min-Ho; Baek, Won-Ki

    2015-12-01

    Juglone (5-hydroxy-1,4-naphthoquinone) is a major chemical constituent of Juglans mandshruica Maxim. Recent studies have demonstrated that juglone exhibits anti-cancer, anti-bacterial, anti-viral, and anti-parasitic properties. However, its effect against Acanthamoeba has not been defined yet. The aim of this study was to investigate the effect of juglone on Acanthamoeba. We demonstrate that juglone significantly inhibits the growth of Acanthamoeba castellanii at 3-5 μM concentrations. Juglone increased the production of reactive oxygen species (ROS) and caused cell death of A. castellanii. Inhibition of ROS by antioxidant N-acetyl-l-cysteine (NAC) restored the cell viability. Furthermore, our results show that juglone increased the uptake of mitochondrial specific dye. Collectively, these results indicate that ROS played a significant role in the juglone-induced cell death of Acanthamoeba.

  5. Temperature-dependent parasitic relationship between Legionella pneumophila and a free-living amoeba (Acanthamoeba castellanii).

    Science.gov (United States)

    Ohno, Akira; Kato, Naoyuki; Sakamoto, Ryota; Kimura, Soichiro; Yamaguchi, Keizo

    2008-07-01

    We analyzed the effects of temperature on the interaction of Legionella pneumophila with Acanthamoeba castellanii. At amoeba. At low temperatures, A. castellanii seems to eliminate L. pneumophila by encystation and digestion.

  6. Osteomielitis esternal y escrofuloderma por vacuna BCG.

    Directory of Open Access Journals (Sweden)

    Ivohne Fernanda Corrales

    2003-06-01

    Full Text Available La vacuna BCG se ha usado en todo el mundo desde principios del siglo XX para la prevención de la tuberculosis. Se describe el caso de una niña de 13 meses de edad, previamente sana, que consultó por una masa esternal. El estudio radiológico mostró erosión perióstica. La lesión fue resecada y en la histopatología se encontró una reacción inflamatoria crónica con granulomas caseificantes con compromiso óseo y cutáneo. Se realizó una amplificación por PCR con iniciadores específicos de Mycobacterium tuberculosis del ADN obtenido a partir del tejido incluido en parafina, cuyo resultado fue negativo. Los antecedentes de vacunación con BCG, la aparición de este tipo de granulomas y la ausencia de ADN de M. tuberculosis en el tejido resecado apoyan el diagnóstico de osteomielitis esternal y escrofuloderma por BCG. La osteomielitis es una complicación infrecuente de la vacunación por BCG, que puede presentarse especialmente en pacientes inmunosuprimidos. La evolución clínica de la paciente no ha demostrado ninguna forma de inmunodeficiencia.

  7. Acanthamoeba keratitis - A six year epidemiological review from a tertiary care eye hospital in South India

    OpenAIRE

    Manikandan P; Bhaskar M; Revathy R; John R.; Narendran V; Panneerselvam K

    2004-01-01

    PURPOSE: This study analyses the prevalence, demography, predisposing factors and seasonal variation of Acanthamoeba keratitis. METHODS: A retrospective review of all cases presenting with keratitis at the cornea clinic, Aravind Eye Hospital, Coimbatore, from August 1997 to July 2003, was done for screening patients with a provisional diagnosis of Acanthamoeba keratitis. Their records were further analyzed for microbiological details. Cases with culture proven Acanthamoeba keratitis were incl...

  8. Identification of 18S ribosomal DNA genotype of Acanthamoeba from hot spring recreation areas in the central range, Taiwan

    Science.gov (United States)

    Hsu, Bing-Mu; Ma, Po-Hua; Liou, Tai-Sheng; Chen, Jung-Sheng; Shih, Feng-Cheng

    2009-04-01

    SummaryAcanthamoeba is a free-living amoebae ubiquitous to aquatic environments. Within the genus a few species are recognized as opportunistic potential human pathogens, which cause granulomatous amoebic encephalitis (GAE) and keratitis. Infections of keratitis are frequently reported through wearing lens while swimming in the non-disinfected aquatic environment. Contaminations in hot tubs, spas and public baths are also possible. As a result, in this study, we identified Acanthamoeba based on the PCR amplification with a genus-specific primer pair and investigated the distribution of Acanthamoeba at five hot spring recreation areas in central range, Taiwan. We gathered data on factors potentially associated with the pathogen's distribution, including various sampling sites, aquatic environment, physical and microbiological water quality parameters. Spring water was collected from 55 sites and Acanthamoeba was detected in 9 (16.4%). The most frequently detected was Acanthamoeba griffini, followed by Acanthamoeba jacobsi. Legionella were detected in 18 (32.7%) of the sites sampled in this study. The species of Legionella identified included Legionella pneumophila serotype 6, serotype 1, and Legionella erythra. Overall, 9.1% of the samples contained both Acanthamoeba and Legionella. The prevalence of Acanthamoeba was contrary to the levels of microbiological indicators recommended by Taiwan CDC, and no significant differences (Mann-Whitney U test, P Acanthamoeba and water quality parameters. Results of this survey confirm the existence of Acanthamoeba in Taiwan spring recreation areas. Acanthamoeba, the organism responsible for the majority of Acanthamoeba keratitis and can serve as vehicles for facultative pathogens, should be considered a potential threat for health associated with human activities in spring recreation areas of Taiwan.

  9. Intracellular Localization and Trafficking of Serine Proteinase AhSub and Cysteine Proteinase AhCP of Acanthamoeba healyi

    OpenAIRE

    Moon, E.-K.; Lee, S.-T.; Chung, D.-I.; Kong, H.-H.

    2006-01-01

    Proteinases have been proposed to play important roles in pathogenesis and various biologic actions in Acanthamoeba. Although genetic characteristics of several proteases of Acanthamoeba have been reported, the intracellular localization and trafficking of these enzymes has yet to be studied. In the present study, we analyzed the intracellular localization and trafficking of two proteinases, AhSub and AhCP, of Acanthamoeba healyi by transient transfection. Full-length AhSub-enhanced green flu...

  10. Acanthamoeba castellanii induces host cell death via a phosphatidylinositol 3-kinase-dependent mechanism

    OpenAIRE

    Sissons, J.; Kim, K.S.; Stins, M; Jayasekera, S.; Alsam, S.; Khan, Naveed Ahmed

    2005-01-01

    Granulomatous amoebic encephalitis due to Acanthamoeba castellanii is a serious human infection with fatal consequences, but it is not clear how the circulating amoebae interact with the blood-brain barrier and transmigrate into the central nervous system. We studied the effects of an Acanthamoeba encephalitis isolate belonging to the T1 genotype on human brain microvascular endothelial cells, which constitute the blood-brain barrier. Using an apoptosis-specific enzyme-linked immunosorbent as...

  11. Cloning and characterization of a novel mannose-binding protein of Acanthamoeba.

    Science.gov (United States)

    Garate, Marco; Cao, Zhiyi; Bateman, Erik; Panjwani, Noorjahan

    2004-07-09

    Acanthamoebae produce a painful, blinding infection of the cornea. The mannose-binding protein (MBP) of Acanthamoeba is thought to play a key role in the pathogenesis of the infection by mediating the adhesion of parasites to the host cells. We describe here the isolation and molecular cloning of Acanthamoeba MBP. The MBP was isolated by chromatography on the mannose affinity gel. Gel filtration experiments revealed that the Acanthamoeba lectin is a approximately 400-kDa protein that is constituted of multiple 130-kDa subunits. Cloning and sequencing experiments indicated that the Acanthamoeba MBP gene is composed of 6 exons and 5 introns that span 3.6 kb of the amoeba genome and that MBP cDNA codes for a precursor protein of 833 amino acids. That the cloned cDNA encodes authentic MBP was demonstrated by showing that: (i). recombinant MBP possesses mannose binding activity, and (ii). polyclonal antibodies prepared against Acanthamoeba MBP bound to the recombinant protein. Sequence analysis revealed that the MBP contains a large N-terminal extracellular domain, a transmembrane domain, and a short C-terminal cytoplasmic domain. Despite extensive BLAST searches using the MBP sequence, no significant matches were retrieved. The most striking feature of the Acanthamoeba MBP sequence is the presence of a cysteine-rich region containing 14 CXCXC motifs within the extracellular domain. In summary, we have isolated, cloned, and characterized a novel MBP from Acanthamoeba. Because the presence of antibodies to MBP in tears provides protection against infection, the availability of the MBP cDNA sequence and rMBP should help develop: (i). a tear-based test to identify individuals who are at risk of developing the keratitis and (ii). strategies to immunize high-risk individuals.

  12. Vectorial role of Acanthamoeba in Legionella propagation in water for human use.

    Science.gov (United States)

    Magnet, A; Peralta, R H S; Gomes, T S; Izquierdo, F; Fernandez-Vadillo, C; Galvan, A L; Pozuelo, M J; Pelaz, C; Fenoy, S; Del Águila, C

    2015-02-01

    Legionella spp. is the causative agent of Legionnaires' disease and is transmitted through aerosols emanating from man-made water systems. Legionella resistance to water treatments has been related to its association with environmental amoebae such as Acanthamoeba. Due to the high presence of this protozoon in Spain and the high rate of notification of Legionnaires' disease of this country, the aims of this work were to study the coexistence of these bacteria and protozoa in water as well as their interaction. The usefulness of Acanthamoeba co-culture for the isolation of environmental Legionella was also studied. For this purpose, 70 water samples were collected in 2011 from three Drinking Water Treatment Plants, three Wastewater Treatment Plants and five Natural Pools in Spain. Acanthamoeba was found by PCR in 87.1% (61/70) samples and, by culture in 85.7% (60/70) samples. Legionella was detected by PCR in 58.6% (41/70) of water samples, in 5.7% (4/70) by agar culture and 75.7% (53/70) by Acanthamoeba co-culture. From the 54 Acanthamoeba water isolates, Legionella was detected in 43 of them independently of Acanthamoeba's genotype (T3, T4 and T11). Legionella feeleii, Legionella birminghamiensis, Legionella gresilensis/berliardensis, Legionella fairfieldensis, Legionella drozanski and Legionella falloni were identified. In conclusion, our results showed that environmental Acanthamoeba is infected by Legionella to a high percentage, and due to its ubiquity, high resistance and its pathogenic potential per se, new methods for its elimination should be studied. Also, the high effectivity of Acanthamoeba co-culture for Legionella detection has been shown.

  13. Reevaluation of an Acanthamoeba Molecular Diagnostic Algorithm following an Atypical Case of Amoebic Keratitis.

    Science.gov (United States)

    Lau, Rachel; Cunanan, Marlou; Jackson, Jonathan; Ali, Ibne Karim M; Chong-Kit, Ann; Gasgas, Jason; Tian, Jinfang; Ralevski, Filip; Boggild, Andrea K

    2015-10-01

    Amoebic keratitis (AK) is a potentially blinding infection, the prompt diagnosis of which is essential for limiting ocular morbidity. We undertook a quality improvement initiative with respect to the molecular detection of acanthamoebae in our laboratory because of an unusual case of discordance. Nine ATCC strains of Acanthamoeba and 40 delinked, biobanked, surplus corneal scraping specimens were analyzed for the presence of acanthamoebae with four separate real-time PCR assays. The assay used by the Free-Living and Intestinal Amebas Laboratory of the CDC was considered the reference standard, and the performance characteristics of each individual assay and pairs of assays were calculated. Outcome measures were sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Of 49 included specimens, 14 (28.6%) were positive by the gold standard assay, and 35 (71.4%) were negative. The sensitivities of the individual assays ranged from 64.3% to 92.9%, compared to the gold standard, while the specificities ranged from 88.6% to 91.4%. The PPVs and NPVs ranged from 69.2% to 78.6% and from 86.1% to 96.9%, respectively. Combinations of assay pairs led to improved performance, with sensitivities ranging from 92.9% to 100% and specificities ranging from 97.1% to 100%. ATCC and clinical strains of Acanthamoeba that failed to be detected by certain individual assays included Acanthamoeba castellanii, Acanthamoeba culbertsoni, and Acanthamoeba lenticulata. For three clinical specimens, false negativity of the gold standard assay could not be excluded. Molecular diagnostic approaches, especially combinations of highly sensitive and specific assays, offer a reasonably performing, operator-independent, rapid strategy for the detection of acanthamoebae in clinical specimens and are likely to be more practical than either culture or direct microscopic detection.

  14. Campylobacter jejuni Actively Invades the Amoeba Acanthamoeba polyphaga and Survives within Non Digestive Vacuoles

    OpenAIRE

    Jenny Olofsson; Diana Axelsson-Olsson; Lars Brudin; Björn Olsen; Patrik Ellström

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81–176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furt...

  15. Acanthamoeba polyphaga mimivirus prevents amoebal encystment-mediating serine proteinase expression and circumvents cell encystment.

    Science.gov (United States)

    Boratto, Paulo; Albarnaz, Jonas Dutra; Almeida, Gabriel Magno de Freitas; Botelho, Lucas; Fontes, Alide Caroline Lima; Costa, Adriana Oliveira; Santos, Daniel de Assis; Bonjardim, Cláudio Antônio; La Scola, Bernard; Kroon, Erna Geessien; Abrahão, Jônatas Santos

    2015-03-01

    Acanthamoeba is a genus of free-living amoebas distributed worldwide. Few studies have explored the interactions between these protozoa and their infecting giant virus, Acanthamoeba polyphaga mimivirus (APMV). Here we show that, once the amoebal encystment is triggered, trophozoites become significantly resistant to APMV. Otherwise, upon infection, APMV is able to interfere with the expression of a serine proteinase related to amoebal encystment and the encystment can no longer be triggered.

  16. The role of Src kinase in the biology and pathogenesis of Acanthamoeba castellanii

    OpenAIRE

    Siddiqui Ruqaiyyah; Iqbal Junaid; Maugueret Marie-josée; Khan Naveed

    2012-01-01

    Abstract Background Acanthamoeba species are the causative agents of fatal granulomatous encephalitis in humans. Haematogenous spread is thought to be a primary step, followed by blood–brain barrier penetration, in the transmission of Acanthmaoeba into the central nervous system, but the associated molecular mechanisms remain unclear. Here, we evaluated the role of Src, a non-receptor protein tyrosine kinase in the biology and pathogenesis of Acanthamoeba. Methods Amoebistatic and amoebicidal...

  17. Acanthamoeba and Stenotrophomonas maltophilia keratitis with fungal keratitis in the contralateral eye

    OpenAIRE

    2010-01-01

    Thomas F Mauger, Rebecca Ann Kuennen, Reynell Harder Smith, William SawyerDepartment of Ophthalmology, The Ohio State University, Columbus, OH, USAPurpose: The purpose of this study is to describe the diagnosis, course, and outcome of a case of Acanthamoeba and Stenotrophomonas keratitis with a fungal keratitis in the contralateral eye.Methods: A case of Acanthamoeba and Stenotrophomonas keratitis was diagnosed with confocal microscopy and cultures with confocal diagnosis of fungal keratitis ...

  18. Detection of Acanthamoeba and Toxoplasma in River Water Samples by Molecular Methods in Iran.

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Mahmoudi

    2015-06-01

    Full Text Available Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples.A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively.Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites.The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oocystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran.

  19. Confoscan: An ideal therapeutic Aid and screening tool in acanthamoeba keratitis

    Directory of Open Access Journals (Sweden)

    Nadia Al Kharousi

    2012-01-01

    Full Text Available Although present worldwide, Acanthamoeba keratitis (AK is a rare condition. It is a protozoal infection of the eye that is generally caused by wearing contaminated contact lenses or lens solutions. Confoscan and confocal scanning laser tomography (CSLT are in vivo noninvasive diagnostic tools which provide high definition images of corneal microstructures. Laser in situ keratomileusis (LASIK is a very common refractive surgery. We report a case series in which the first patient had contact lens induced Acanthamoeba keratitis with corneal epitheliopathy that was unresponsive to conservative treatment. Epithelial debridement was performed based on confoscan findings which confirmed the presence of Acanthamoeba cysts. Subsequently, the cornea re-epithelialized over two days. Another patient had CSLT prior to the LASIK which showed stromal cyst-like structures suggestive of Acanthamoeba keratitis. Four months after medical therapy, repeat CSLT was negative for Acanthamoeba cysts. Third patient was diagnosed with Acanthamoeba infection after undergoing lamellar keratoplasty. CSLT should be used as a screening procedure prior to any corneal refractive surgery to detect and treat protozoal and other infections preoperatively.

  20. Granulomatous inflammation in acanthamoeba keratitis: An immunohistochemical study of five cases and review of literature

    Directory of Open Access Journals (Sweden)

    Vemuganti G

    2005-01-01

    Full Text Available Purpose: Acanthamoeba keratitis usually presents as a necrotizing stromal inflammation. We report a rare presentation of granulomatous inflammation in Acanthamoeba keratitis Methods: Retrospective clinico-pathologic case series. Results: Five corneal tissues (3 corneal buttons, 2-eviscerated contents from patients suffering from severe Acanthamoeba keratitis not responding to anti-Acanthamoeba treatment, revealed a florid granulomotous inflammation with multinucleated giant cells in the posterior stroma and around Descemet′s membrane. Phagocytosed parasites were noted within the giant cells. Vascularization of the corneal stroma was noted in two cases. Immunophenotyping revealed a predominance of T lymphocytes and macrophages. Clinically, four of five cases had shown features of limbal and scleral involvement. Conclusion: Granulomatous inflammation in the posterior corneal stroma, is not an uncommon finding in Acanthamoeba keratitis and could possibly be immune-mediated, contributing to persistence and progression of disease. Clinical Relevance: Presence of granulomatous inflammation in Acanthamoeba keratitis, in most cases is associated with limbal and scleral involvement and therefore could be considered as one of the poor prognostic markers. Further studies are required to ascertain the specific clinical features and appropriate management strategies in these cases.

  1. Health effects of Acanthamoeba spp. and its potential for waterborne transmission.

    Science.gov (United States)

    Nwachuku, Nena; Gerba, Charles P

    2004-01-01

    Risk from Acanthamoeba keratitis is complex, depending upon the virulence of the particular strain, exposure, trauma, or other stress to the eye, and host immune response. Bacterial endosymbionts may also play a factor in the pathogenicity of Acanthamoeba. Which factor(s) may be the most important is not clear. The ability of the host to produce IgA antibodies in tears may be a significant factor. The immune response of the host is a significant risk factor for GAE infection. If so, then a certain subpopulation with an inability to produce IgA in the tears may be at greatest risk. There was no sufficient data on the occurrence or types of Acanthamoeba in tapwater in the U.S. Published work on amoebal presence in tapwater does not provide information on the type of treatment the water received or the level of residual chlorine. Assessment of the pathogenicity by cell culture and molecular methods of Acanthamoeba in tapwater would also be useful in the risk assessment process for drinking water. The possibility that Acanthamoeba spp. might serve as vectors for bacterial infections from water sources also should be explored. The bacterial endosymbionts include an interesting array of pathogens such as Vibrio cholerae and Legionella pneumophila, both of which are well recognized waterborne/water-based pathogens. Work is needed to determine if control of Acanthamoeba spp. is needed to control water-based pathogens in water supplies.

  2. Presence of Acanthamoeba spp.in water purification plants in southern England

    Institute of Scientific and Technical Information of China (English)

    Shanmuganathan V; Khan NA

    2009-01-01

    Objective:To identify the prevalence of Acanthamoeba in drinking water treatment plants during the course of the purification processes.Methods:Samples were taken from two drinking water purification plants and moni-tored for the presence of Acanthamoeba in order to estimate the removal capacity of treatment methods em-ployed.Water samples were collected at each step in the purification,during the one year survey,and ana-lysed for the presence of Acanthamoeba spp.by plating on bacterial-seeded plates.Results:The results showed that amoebae were present in surface raw waters in 100 % of the samples tested.Acanthamoeba spp.were iso-lated from 71 % and 57 % of the water samples collected from post flat-bottom clarifier 1 and post-sedimenta-tion plant respectively.Considering the outflow drinking waters,the removal capacity was 100 % in both puri-fication plants monitored.The occurrence of Acanthamoeba was not associated with seasonality.Conclusion:These findings confirm that water purification plants employing methods of flocculation,sedimentation,and fil-tration in combination with activated charcoal filtration,ozonisation and chlorination exhibited sufficient Acan-thamoeba removal capacity and the presence of amoebae in the tap water may be due to older plumbing,water storage tanks,tap water hygiene,and /or environmental settings.

  3. Comparative analyses of different genetic markers for the detection of Acanthamoeba spp. isolates.

    Science.gov (United States)

    Derda, Monika; Wojtkowiak-Giera, Agnieszka; Hadaś, Edward

    2014-09-01

    Acanthamoeba are widespread free-living amoebae which may cause granulomatous amoebic encephalitis (GAE), keratitis, skin ulcerations and disseminated tissue infection. An important diagnostic and prognostic factor for the treatment of infection is a quick and correct diagnosis of amoebae strains. The aim of our study was to develop a rapid method for detection and identification of pathogenic Acanthamoeba spp. strains from diagnostic material collected from water. In this study we analysed five amplification-based genetic markers (Aca 16S, Ac6/210, GP, JDP, Nelson) used for identification of pathogenic Acanthamoeba spp. strains isolated in water sources in Poland, Iceland and Sweden. Our results demonstrated the presence of pathogenic Acanthamoeba strains in tap water. PCR assay appeared to be a more rapid and sensitive method to detect the presence of amoebae than the limited conventional techniques. Based on our observations, we can confirm that the use of four out of five genetic markers (Aca 16S, Ac 6/210, JDP, GP, Nelson) may be helpful in identification of Acanthamoeba spp. strains, but only one Aca 16S primer pair is a highly specific marker that distinguishes between pathogenic strains of Acanthamoeba and other free-living amoeba families.

  4. Isolation and characterization of Acanthamoeba strains from soil samples in Gran Canaria, Canary Islands, Spain.

    Science.gov (United States)

    Reyes-Batlle, María; Todd, Cheridah D; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Cabello-Vilchez, Alfonso Martín; González, Ana C; Córdoba-Lanús, Elizabeth; Lindo, John F; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-04-01

    Free-living Amoebae of Acanthamoeba genus include non-pathogenic and pathogenic strains that are currently classified in 18 different genotypes, T1-T18. In this study, a survey was carried out to evaluate the presence of Acanthamoeba strains in soil samples collected between 2012 and 2013 in Gran Canaria Island, Canary Islands, Spain. Samples were inoculated onto non-nutrient agar (NNA) plates and were checked for the presence of Acanthamoeba. Identification of Acanthamoeba strains was based on the morphology of the cyst and trophozoite forms. Subsequently, positive samples were cloned for their molecular characterization at the genotype level by sequencing the DF3 region located in the 18S rDNA gene of Acanthamoeba as previously described. Sequencing results revealed the presence of T2, T5 and T4 genotypes within the studied samples. To the best of our knowledge, this is the first report demonstrating the presence of Acanthamoeba in Gran Canaria Island and the first study at the genotype level in the Canary Islands.

  5. Characterisation and differentiation of pathogenic and non-pathogenic Acanthamoeba strains by their protein and antigen profiles.

    Science.gov (United States)

    Walochnik, J; Sommer, K; Obwaller, A; Haller-Schober, E-M; Aspöck, H

    2004-03-01

    Free-living amoebae of the genus Acanthamoeba are the causative agents of Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis. Acanthamoebae occur ubiquitously in the environment and are thus a constant cause of antigenic stimulation. In a previous study we have shown that compared to control sera, AK patients exhibit markedly lower immunoreactivities to whole cell antigen of Acanthamoeba spp. As the pathogenicity of acanthamoebae primarily relies on the excretion of proteins, it was the aim of the present study to investigate the immunoreactivity of metabolic antigen from different Acanthamoeba strains of varying pathogenicity. Three Acanthamoeba strains, one highly pathogenic, one non-pathogenic but thermophilic and one non-thermophilic non-pathogenic, were used for antigen extraction. The antigen was harvested before and after contact with human cells and all strains were tested with AK sera and with sera from healthy individuals. It was shown that the somatic protein profiles of the Acanthamoeba strains correlated to the morphological groups, and that within morphological group II-the group associated with AK-the profiles of the metabolic antigens correlated to strain pathogenicity. Moreover, it was shown that the control sera showed markedly higher immunoreactivities than the sera of the AK patients and that this immunoreactivity was generally higher to the non-pathogenic strains than to the pathogenic strain. Altogether our results once again raise the question of whether there is an immunological predisposition in AK. To our knowledge this is the first study on the immunoreactivity of metabolic antigen of acanthamoebae.

  6. First report of an Acanthamoeba genotype T13 isolate as etiological agent of a keratitis in humans.

    Science.gov (United States)

    Grün, Anna-Lena; Stemplewitz, Birthe; Scheid, Patrick

    2014-06-01

    Several strains of free-living amoebae (FLA) belonging to the genus Acanthamoeba are able to cause a painful sight-threatening disease of the cornea designated as Acanthamoeba keratitis (AK). In this case report, a 22-year-old woman, wearer of soft contact lenses, was treated after the initial examination, and follow-up laboratory results led to the diagnosis of Acanthamoeba keratitis. The patient recovered under the targeted therapy, demonstrating that the acanthamoebae were the etiological agents of the keratitis in this case. The acanthamoebae belonged morphologically to group II. Genotyping of the causative Acanthamoeba strain based on sequences of the PCR amplimer ASA.S1 amplified from 18S ribosomal DNA by using the genus-specific primers JDP1 and JDP2 followed. The phylogenetic comparison of ASA.S1 confirmed that the isolated Acanthamoeba strain is closely related to genotype T13 supported by pairwise sequence identities of 97.1-98.0% and bootstrap support of 980 replicates with reference sequences of genotype T13. These results regarding the Acanthamoeba keratitis-causing isolate KaBo expands the number of known pathogenic genotypes to 12. To our knowledge, this is the first report of a T13 Acanthamoeba genotype being associated with keratitis in humans.

  7. Use of 18S rRNA Gene-Based PCR Assay for Diagnosis of Acanthamoeba Keratitis in Non-Contact Lens Wearers in India

    OpenAIRE

    Pasricha, Gunisha; Sharma, Savitri; Garg, Prashant; Aggarwal, Ramesh K.

    2003-01-01

    Identification of Acanthamoeba cysts and trophozoites in ocular tissues requires considerable expertise and is often time-consuming. An 18S rRNA gene-based PCR test, highly specific for the genus Acanthamoeba, has recently been reported in the molecular diagnosis of Acanthamoeba keratitis. This PCR assay was compared with conventional microbiological tests for the diagnosis of Acanthamoeba keratitis. In a pilot study, the PCR conditions with modifications were first tested on corneal scraping...

  8. Isolation of Acanthamoeba isolates belonging to T2, T3, T4 and T7 genotypes from environmental samples in Ankara, Turkey

    OpenAIRE

    Kiliç, A.; Tanyuksel, M.; Sissons, J.; Jayasekera, S.; Khan, Naveed Ahmed

    2004-01-01

    Acanthamoeba keratitis is a blinding infection that is becoming increasingly important in human health. Early diagnosis is a prerequisite for successful treatment and requires identification of Acanthamoeba at the genotypic level. The genus Acanthamoeba consists of both pathogenic and non-pathogenic species and has been recently classified into 13 different genotypes, T1-T12 and T14. More importantly, 95% of Acanthamoeba isolates that produce keratitis belong to T4 genotypes. In this study, w...

  9. Selenio en los pacientes críticos con Respuesta Inflamatoria Sistémica Selenium in critically ill patients with systemic inflammatory response

    Directory of Open Access Journals (Sweden)

    W. Manzanares Castro

    2007-06-01

    Full Text Available El Selenio es un micronutriente esencial para el hombre. Los estados de Respuesta Inflamatoria Sistémica (RIS y Disfunción Orgánica Múltiple (DOM tales como la sepsis severa, el trauma grave, la pancreatitis severa y quemaduras críticas se caracterizan por la existencia de estrés oxidativo. En este contexto, la depleción del estado nutricional de Selenio es una característica distintiva; la misma se traduce por una reducción de los niveles séricos de Selenio y de las diferentes selenoenzimas, en especial la Glutatión Peroxidasa y la Selenoproteína P. Estas selenoproteínas antioxidantes cumplen un rol protectivo frente a la lipoperoxidación de las membranas celulares participando además en el proceso de regulación de la respuesta inflamatoria. Ha sido demostrada la existencia de una correlación directa entre el grado de depleción de Selenio y la severidad de la enfermedad crítica. En los últimos años, numerosos estudios clínicos han investigado los efectos de la suplementación de Selenio en pacientes críticos con RIS -DOM. Esta estrategia terapéutica sería capaz de mejorar los resultados optimizando el pronóstico de la enfermedad crítica. En tal sentido, algunos pequeños estudios clínicos han demostrado una disminución del número de complicaciones infecciosas y de la estadía hospitalaria en los grupos de pacientes críticos suplementados con Selenio. Sin embargo, hasta el momento actual no existen datos concluyentes que permitan definitivamente afirmar la existencia de una reducción significativa de la mortalidad con el uso de Selenio en Terapia Intensiva. El objetivo del presente artículo de revisión es estudiar el metabolismo y estado nutricional del Selenio, evaluar el rol de las selenoenzimas durante la enfermedad crítica y repasar los estudios de suplementación de Selenio como estrategia antioxidante en Terapia Intensiva analizando por último la evidencia actual. Las recomendaciones actuales parecen

  10. Cirugía bariátrica en enfermedad inflamatoria intestinal: presentación de un caso clínico y revisión de la literatura

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    Carmen Tenorio Jiménez

    2013-06-01

    Full Text Available La Enfermedad Inflamatoria Intestinal (EII raramente se asocia a obesidad, ya que la malabsorción es una característica frecuente de este grupo de patologías (1. Sin embargo, algunos pacientes pueden padecer obesidad mórbida asociada a complicaciones y refractaria a tratamiento dietético y beneficiarse de la cirugía bariátrica. Incluso se ha postulado que podría producirse una mejoría de la EII al disminuir los marcadores inflamatorios tras la cirugía (2. No obstante, los pacientes pueden experimentar mayor incidencia de complicaciones tras la cirugía en el contexto de terapias inmunosupresoras y agravamiento de la malabsorción previa. Por ello, si se realiza la cirugía, la cuidadosa selección de los pacientes y la individualización de la técnica a realizar son imprescindibles. Presentamos una paciente diagnosticada de Colitis Ulcerosa que presenta desnutrición proteica severa tras cirugía bariátrica tipo derivación bilio-pancreática y realizamos una revisión de la literatura disponible.

  11. Comportamiento de variables epidemiológicas en la neuropatía óptica inflamatoria infecciosa. Centro Oftalmológico de Holguín. 2009

    Directory of Open Access Journals (Sweden)

    Enma Estrella de la Torre-Rodríguez

    2012-01-01

    Full Text Available Abordó un estudio descriptivo del comportamiento de algunas variables epidemiológicas en un universo de 18 pacientes con neuropatía óptica inflamatoria en el Centro Oftalmológico de Holguín de enero a diciembre de 2009. La muestra estuvo representada por 12 pacientes con etiología infecciosa. El mayor número de pacientes se incluyó en el grupo de edad de 15 a 29 años. Las ocupaciones de riesgo más frecuentes fueron los estudiantes, las amas de casa y criadores de palomas. Los perros, gatos y cerdos fueron los animales más frecuentes en contacto con los casos. La etiología parasitaria representó el grupo más frecuente como causa de la neuropatía y el agente que predominó fue Bartonella henselae. Se recomendó realizar estudios que incluyan confirmación serológica y/o del agente causal en este grupo de enfermedades.

  12. Acanthamoeba castellanii: morphological analysis of the interaction with human cornea.

    Science.gov (United States)

    Omaña-Molina, Maritza; González-Robles, Arturo; Salazar-Villatoro, Lizbeth Iliana; Cristóbal-Ramos, Ana Ruth; González-Lázaro, Mónica; Salinas-Moreno, Edmundo; Méndez-Cruz, Rene; Sánchez-Cornejo, Manuel; De la Torre-González, Enrique; Martínez-Palomo, Adolfo

    2010-09-01

    The present study demonstrates that when Acanthamoeba castellanii trophozoites are co-cultivated with isolated human corneas, the amoeba can be invasive and cause damage to the intact corneal epithelium without the requirement of previous corneal abrasion. After adhesion, A. castellanii trophozoites migrate between cells forming bumps on the corneal cell layers and reaching Bowman s membrane in 3h, although no evidence of cell damage was observed until the phagocytic process was detected. Likewise, conditioned medium produced damage to the corneal cells that was proportional to the time of incubation, but this cytophatic effect involved only the most superficial layer of the human cornea and was not enough to explain amoebic invasion of Bowman s membrane. As a result of our observations, we suggest that the mechanical action of the trophozoites and phagocytosis of corneal cells during the process of corneal invasion are more important than previously suggested.

  13. Pneumonia in mice inoculated experimentally with Acanthamoeba polyphaga mimivirus.

    Science.gov (United States)

    Khan, Mohammad; La Scola, Bernard; Lepidi, Hubert; Raoult, Didier

    2007-01-01

    In an attempt to elucidate whether Acanthamoeba polyphaga mimivirus (APM) was pathogenic (i.e., induces histological evidence of inflammation or tissue destruction or both) in mammalian host, we inoculated adult BALB/c (n=6) and C57BL/6 (n=6) mice with APM via intracardiac route. C57BL/6 mice developed histopathological features of pneumonia by post-inoculation day (PID) 3 and BALB/c mice, by PID7. The histopathological features of pneumonia, characterized by the presence of thickened alveolar walls with cellular infiltrates comprising mononuclear leukocytes, macrophages and lymphocytes, diffuse alveolar damages with the formation of hyaline membrane and erythrocytes in the alveolar lumen, were most marked by PID 7. We conclude that APM is pathogenic in mice in experimental conditions.

  14. Interactions of some common pathogenic bacteria with Acanthamoeba polyphaga.

    Science.gov (United States)

    Huws, Sharon A; Morley, Robert J; Jones, Martin V; Brown, Michael R W; Smith, Anthony W

    2008-05-01

    Protozoan grazing is a major trophic pathway whereby the biomass re-enters the food web. Nonetheless, not all bacteria are digested by protozoa and the number known to evade digestion, resulting in their environmental augmentation, is increasing. We investigated the interactions of Bacillus cereus, Enterococcus faecalis, Enteropathogenic Escherichia coli (EPEC), Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and methicillin-sensitive Staphylococcus aureus (MSSA), with the amoeba, Acanthamoeba polyphaga. There was evidence of predation of all bacterial species except L. monocytogenes and S. aureus, where extracellular numbers were significantly higher when cultured with amoebae compared with growth in the absence of amoebae. Intracellular growth kinetic experiments and fluorescent confocal microscopy suggest that S. aureus survived and may even multiply within A. polyphaga, whereas there was no apparent intra-amoebal replication of L. monocytogenes and higher numbers were likely sustained on metabolic waste products released during coculture.

  15. QUERATOPATÍA CRISTALINA ATÍPICA POR CANDIDA PARAPSILOSIS

    Directory of Open Access Journals (Sweden)

    Raquel Martínez Chico

    2013-10-01

    Full Text Available Caso clínicoMujer de 91 años con antecedentes de  queratoplastia penetrante excéntrica. Un año después presentó una gran reacción inflamatoria y un infiltrado corneal de aspecto arboriforme. Se inició tratamiento con vancomicina intravenosa y vancomicina y amikacina intravítreas, tópicas e intracamerulares y se obtuvieron muestras de humor acuoso y del infiltrado corneal mediante arrastre con aguja de safil de 8/0 con resultado microbiológico positivo para Candida Parapsilosis. Se inició entonces tratamiento con voriconazol y trimetroprim-sulfametoxazol tópicos con respuesta favorable. DiscusiónEl caso que presentamos es atípico puesto que encontramos una intensa reacción inflamatoria; sin embargo tanto el aspecto biomicroscópico de la cornea, como el resultado del análisis microbiológico y la respuesta positiva al tratamiento apoyan el diagnóstico de queratopatía cristalina por Candida Parapsilosis. Normal 0 21 false false false ES X-NONE X-NONE

  16. Estudio Transversal sobre la Relación entre Neuroticismo y Curso Clínico en Pacientes con Enfermedades Inflamatorias Intestinales

    Directory of Open Access Journals (Sweden)

    Juan Carlos Fernández Méndez

    2010-01-01

    Full Text Available En el presente trabajo se analiza la relación que existe entre neuroticismo y diversas condiciones médicas que hacen referencia al curso clínico de las enfermedades inflamatorias intestinales. Para desarrollar la investigación se tomaron medidas en neuroticismo mediante el Inventario de Personalidad de Eysenck a 106 participantes diagnosticados de colitis ulcerosa y enfermedad de Crohn. Los resultados reflejaron que la muestra global de pacientes con enfermedades inflamatorias intestinales mostraba valores en neuroticismo que se hallaban dentro de la normalidad; sin embargo, se pudo verificar que los valores diferían dependiendo del curso clínico de la patología. El estudio sugiere que las variaciones en neuroticismo son considerables cuando se analizan los datos de la muestra general y cuando se tienen en cuenta diversas condiciones del curso clínico.

  17. Bowman’s layer encystment in cases of persistent Acanthamoeba keratitis

    Directory of Open Access Journals (Sweden)

    Yokogawa H

    2012-08-01

    Full Text Available Hideaki Yokogawa,1 Akira Kobayashi,1 Natsuko Yamazaki,1 Yasuhisa Ishibashi,2 Yosaburo Oikawa,3 Masaharu Tokoro,4 Kazuhisa Sugiyama11Department of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, 2Department of Ophthalmology, East Washinomiya Hospital, Kuki, 3Department of Medical Zoology, Kanazawa Medical University, Kahoku, 4Department of Parasitology, Kanazawa University Graduate School of Medical Science, Kanazawa, JapanBackground: The purpose of this study was to report Acanthamoeba encystment in Bowman’s layer in Japanese cases of persistent Acanthamoeba keratitis (AK.Methods: Laser confocal microscopic images of the cornea were obtained in vivo from 18 consecutive eyes from 17 confirmed AK patients. Retrospectively, 14 cases treated over 4 months were categorized as a nonpersistent group and three cases that required prolonged therapy for more than 6 months were categorized as a persistent group. Clinical outcomes based on final best-corrected visual acuity were retrospectively analyzed, and selected confocal images were evaluated qualitatively for abnormal findings.Results: The final best-corrected visual acuity was significantly lower (P < 0.01 for patients in the persistent group compared with that in the nonpersistent group. At the initial visit, in vivo confocal microscopy demonstrated Acanthamoeba cysts exclusively in the epithelial layer in both the nonpersistent group (80% and the persistent group (100%. At a subsequent follow-up visit, numerous Acanthamoeba cysts were observed in the epithelial cell layer and in Bowman’s layer in all patients with persistent AK, but Acanthamoeba cysts were undetectable in all cases with nonpersistent AK tested.Conclusion: Invasion of cysts into Bowman’s layer was characteristically observed in patients with persistence of AK. This finding suggests that invasion of Acanthamoeba cysts into Bowman’s layer may be a useful predictor for a persistent clinical

  18. Pathogenic strains of Acanthamoeba are recognized by TLR4 and initiated inflammatory responses in the cornea.

    Directory of Open Access Journals (Sweden)

    Hassan Alizadeh

    Full Text Available Free-living amoebae of the Acanthamoeba species are the causative agent of Acanthamoeba keratitis (AK, a sight-threatening corneal infection that causes severe pain and a characteristic ring-shaped corneal infiltrate. Innate immune responses play an important role in resistance against AK. The aim of this study is to determine if Toll-like receptors (TLRs on corneal epithelial cells are activated by Acanthamoeba, leading to initiation of inflammatory responses in the cornea. Human corneal epithelial (HCE cells constitutively expressed TLR1, TLR2, TLR3, TLR4, and TLR9 mRNA, and A. castellanii upregulated TLR4 transcription. Expression of TLR1, TLR2, TLR3, and TLR9 was unchanged when HCE cells were exposed to A. castellanii. IL-8 mRNA expression was upregulated in HCE cells exposed to A. castellanii. A. castellanii and lipopolysaccharide (LPS induced significant IL-8 production by HCE cells as measured by ELISA. The percentage of total cells positive for TLR4 was higher in A. castellanii stimulated HCE cells compared to unstimulated HCE cells. A. castellanii induced upregulation of IL-8 in TLR4 expressing human embryonic kidney (HEK-293 cells, but not TLR3 expressing HEK-293 cells. TLR4 neutralizing antibody inhibited A. castellanii-induced IL-8 by HCE and HEK-293 cells. Clinical strains but not soil strains of Acanthamoeba activated TLR4 expression in Chinese hamster corneas in vivo and in vitro. Clinical isolates but not soil isolates of Acanthamoeba induced significant (P< 0.05 CXCL2 production in Chinese hamster corneas 3 and 7 days after infection, which coincided with increased inflammatory cells in the corneas. Results suggest that pathogenic species of Acanthamoeba activate TLR4 and induce production of CXCL2 in the Chinese hamster model of AK. TLR4 may be a potential target in the development of novel treatment strategies in Acanthamoeba and other microbial infections that activate TLR4 in corneal cells.

  19. El diagnóstico por la imagen de las enfermedades de las glándulas salivales

    OpenAIRE

    Paz Expósito, Francisco

    2002-01-01

    Las lesiones de las glándulas salivales son relativamente frecuentes, presentando una patología tan diversa como la de cualquier otro sistema orgánico. La mayoría de las lesiones son inflamatorias, otras son de origen litiásico, algunas tienen una patogenia incierta que pueden confundirse con neoplasias malignas y, por último, hay un amplio espectro de neoplasias tanto benignas como malignas. La mayoría de los agrandamientos de las glándulas salivales están causados por procesos inflamator...

  20. Intoxicación por administración parenteral de insecticidas organoclorados e hidrocarburos aromáticos

    Directory of Open Access Journals (Sweden)

    F.J. Boluda

    2003-01-01

    Full Text Available La intoxicación por vía parenteral debida a insecticidas organoclorados e hidrocarburos aromáticos, es muy poco frecuente. La toxicidad comporta efectos locales y sistémicos que pueden llegar a comprometer la vida del paciente. Se presenta el caso clínico de un varón que desarrolló una importante reacción local necrótico-inflamatoria, con leve afectación sistémica hepática y renal, tras administrarse los mencionados productos por vía parenteral en una tentativa de suicidio

  1. Pharmacological Nutrition in Inflammatory Bowel Diseases Nutrición farmacológica en las enfermedades inflamatorias del instestino

    Directory of Open Access Journals (Sweden)

    F. G. Campos

    2003-04-01

    Full Text Available Inflammatory Bowel Diseases - ulcerative colitis and Crohn’s disease- are chronic gastrointestinal inflammatory diseases of unknown etiology. Decreased oral intake, malabsorption, accelerated nutrient losses, increased requirements, and drug-nutrient interactions cause nutritional and functional deficiencies that require proper correction by nutritional therapy. The goals of the different forms of nutritional therapy are to correct nutritional disturbances and to modulate inflammatory response, thus influencing disease activity. Nutritional intervention may improve outcome in certain individuals; however, because of the costs and complications of such therapy, careful selection is warranted. Total parenteral nutrition has been used to correct and prevent nutritional disturbances and to promote bowel rest during active disease, mainly in cases of digestive fistulae with a high output. Its use should be reserved for patients who cannot tolerate enteral nutrition. Enteral nutrition is effective in inducing clinical remission of disease in adults and promoting growth in children. Recent research has focused on the use of specific nutrients as primary treatment agents. Although some reports have indicated that glutamine, short-chain fatty acids, antioxidants and immunonutrition with omega-3 fatty acids are an important therapeutic alternative in the management of inflammatory bowel diseases, the beneficial reported effects have yet to be translated into the clinical practice. The real efficacy of these nutrients still need further evaluation through prospective and randomized trials.Las enfermedades inflamatorias del intestino -colitis ulcerosa y enfermedad de Crohn- son enfermedades crónicas de causa desconocida. La disminución de la ingesta, la malabsorción, la pérdida acelerada de nutrientes, el aumento de los requerimientos y las interacciones entre medicamentos y nutrientes determinan carencias nutricionales y funcionales que obligan a una

  2. Anti-Acanthamoeba IgG, IgM, and IgA immunoreactivities in correlation to strain pathogenicity.

    Science.gov (United States)

    Walochnik, J; Obwaller, A; Haller-Schober, E M; Aspöck, H

    2001-08-01

    Several representatives of the genus Acanthamoeba are known as causative agents of Acanthamoeba keratitis and granulomatous amoebic encephalitis. These occur predominantly in the immunocompromised host, but it is still unclear what primes the amoebae for pathogenicity. The aim of this study was to assess possible immunological differences between a highly pathogenic and a nonpathogenic Acanthamoeba strain. A total of 20 sera, including two sera of Acanthamoeba keratitis patients, were tested for anti-Acanthamoeba IgG, IgM, and IgA immunoreactivities using immunoblotting. All sera were positive for Acanthamoeba, revealing two predominant bands at 29 kDa and at 47 kDa, respectively. Interestingly, IgG and particularly IgA immunoreactivity enabled a clear discrimination between the pathogenic and nonpathogenic strains. Moreover, compared to the control sera, the two sera of Acanthamoeba keratitis patients showed rather weak immunoreactivities and they lacked the 29 kDa and the 47 kDa band in the IgA immunoblot against the pathogenic strain. The results of our study support the assumption that immunological predisposition might also be of importance in Acanthamoeba keratitis.

  3. Morpho-Physiological and Biochemical Criteria of Acantha-moeba spp. Isolated from the Egyptian Aquatic Environment

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    A Mohammed

    2013-06-01

    Full Text Available Background: The free-living amoebae Acanthamoeba spp., have been recognized as etiologic agents of amoebic encephalitis, keratitis, otitis, lung lesions and other skin infections mainly in immuno-compro­mised individuals. In this study, morpho-physiological and biochemical characterization of Acanthamoeba strains isolated from the Egyptian aquatic environment were surveyed.Methods: some Acanthamoeba species were cultivated on non-nutrient agar. Isolated strains of Acantha­moeba were identification based on the morphology of trophic and cyst forms in addition to temperature and osmo-tolerance assays. Biochemical characterization of the isolated amoeba strains was performed using quantitative assay as well as qualitative determination of proteolytic activity in zymograph analysis.Results: Potentially pathogenic Acanthamoeba species were isolated from all of the examined water sources. Colorimetric assays showed protease activity in the heat-tolerant isolates of Acanthamoeba. All pathogenic isolates of Acanthamoeba exhibited higher protease activity than did the non-patho­genic ones. The zymographic protease assays showed various banding patterns for different strains of Acanthamoeba.Conclusio: The incidence and prevalence of the pathogenic Acanthamoeba species in the aquatic environment using parasitological and biochemical diagnostic tools will provide baseline data against which the risk factors associated with waterborne transmission can be identified.

  4. Aftas severas como manifestación de enteropatía inflamatoria

    OpenAIRE

    Roude, A.; Maganini, C.; Tauler, T.

    2015-01-01

    la Celiaquía es un desorden sistémico con base inmunológica causado por la ingesta de gluten y otras proteínas similares que afecta a personas con predisposición genética. La interacción entre el gluten y el sistema inmunológico del paciente puede manifestarse además a diferentes niveles: piel, articulaciones, riñón y en la mucosa oral (estomatitis aftosa redicivante). Descripción del caso: se presenta a la consulta derivada por su odontóloga particular, un paciente de sexo femenino, 39 años ...

  5. Patología infecciosa: vulvovaginitis, enfermedades de transmisión sexual, enfermedad inflamatoria pélvica, abscesos tubo-ováricos Infectious pathology: vulvovaginitis, sexually transmitted diseases, pelvic inflammatory disease, tubo-ovarian abscesses

    Directory of Open Access Journals (Sweden)

    M. Ibarrola

    2009-01-01

    Full Text Available Las enfermedades de transmisión sexual son aquellas en las que la principal vía de infección es el contacto íntimo. Son numerosas las pacientes que acuden a urgencias por esta causa, tanto por la clínica como por las implicaciones sociales. Los síntomas más frecuentes son dolor abdominal bajo, sangrados vaginales, o flujo vaginal excesivo o molesto. Las vulvovaginitis son uno de los problemas principales en la práctica clínica diaria del ginecólogo. La úlcera genital cuya etiología principal es el herpes, seguida de la sífilis y el chancroide incrementa el riesgo para contraer la infección por el VIH y modifica el curso de otras enfermedades de transmisión sexual. La enfermedad pélvica inflamatoria engloba a las infecciones del tracto genital superior femenino. La importancia del diagnóstico precoz y su tratamiento adecuado reside tanto por las complicaciones en la fase aguda como por las secuelas, que incluyen el dolor crónico y la esterilidad.Sexually transmitted diseases are those where the principal path of infection is through intimate contact. Numerous patients attend Accidents and emergencies for this reason, both because of the clinical features and because of social implications. The most frequent symptoms are lower abdominal pain, vaginal bleeding or excessive or troubling vaginal flow. Vulvovaginites are one of the principal problems in the everyday clinical practice of gynaecology. A genital ulcer whose principal aetiology is herpes, followed by syphilis and chancroid, increases the risk of contracting HIV infection and alters the course of other sexually transmitted diseases. Inflammatory pelvic disease encompasses infections of the upper female genital tract. The importance of early diagnosis and suitable treatment is both due to the complications in its acute phase and to its sequels, which include chronic pain and sterility.

  6. Unusual case of methicillin resistant staphylococcus aureus and acanthamoeba keratitis in a non-contact lens wearer from Kashmir, India.

    Science.gov (United States)

    Lone, Rubina; Syed, Khurshid; Abdul, Rashid; Sheikh, Sajjad Ahmed; Shah, Faisal

    2009-01-01

    Acanthamoeba species can cause a chronic, progressive, ulcerative keratitis of the eye, which is not responsive to the usual antimicrobial treatment and is frequently mistaken for stromal herpes keratitis. Acanthamoeba keratitis continues to be a burgeoning and unsolved problem. Although soft contact lens wear is reported as the major risk factor in other parts of the world, reports from India suggest that acanthamoeba keratitis is more common among non-contact lens wearers. An unusual case of coinfection with Acanthamoeba and methicillin resistant staphylococcus aureus (MRSA) as causes of corneal keratitis in a contact lens wearer from Kashmir, India, is reported. Recent findings have shown that MRSA uses amoebae to spread, sidestepping hospital and other protection measures. Cysts of the isolated Acanthamoeba tolerated an incubation temperature of 40°C, indicating a pathogenic species. This case highlights the importance of culture methods in the diagnosis of corneal infection and the choice of treatment regimen.

  7. Acanthamoeba feature a unique backpacking strategy to trap and feed on Listeria monocytogenes and other motile bacteria

    DEFF Research Database (Denmark)

    Doyscher, Dominik; Fieseler, Lars; Dons, Lone Elisabet

    2013-01-01

    Despite its prominent role as an intracellular human pathogen, Listeria monocytogenes normally features a saprophytic lifestyle, and shares many environmental habitats with predatory protozoa. Earlier studies claimed that Acanthamoeba may act as environmental reservoirs for L.?monocytogenes, wher......Despite its prominent role as an intracellular human pathogen, Listeria monocytogenes normally features a saprophytic lifestyle, and shares many environmental habitats with predatory protozoa. Earlier studies claimed that Acanthamoeba may act as environmental reservoirs for L.......?monocytogenes, whereas others failed to confirm this hypothesis. Our findings support the latter and provide clear evidence that L.?monocytogenes is unable to persist in Acanthamoeba castellanii and A.?polyphaga. Instead, external Listeria cells are rapidly immobilized on the surface of Acanthamoeba trophozoites...... that formation of backpacks is not specific for L.?monocytogenes, and independent of bacterial pathogenicity or virulence. Hence, backpacking appears to represent a unique and highly effective strategy of Acanthamoeba to trap and feed on motile bacteria....

  8. Quick survey for detection, identification and characterization of Acanthamoeba genotypes from some selected soil and water samples across Pakistan.

    Science.gov (United States)

    Tanveer, Tania; Hameed, Abdul; Gul, Asma; Matin, Abdul

    2015-01-01

    Acanthamoeba is an opportunistic protozoan pathogen which is widely distributed in nature and plays a pivotal role in ecosystem. Acanthamoeba species may cause blinding keratitis and fatal granulomatous encephalitis involving central nervous system. In this study, we investigated the presence of Acanthamoeba in soil and water resources of Pakistan. Here, Acanthamoeba were recovered on non-nutrient agar plate lawn with E. coli and identified by morphological characteristics of the cyst. Furthermore PCR was performed with genus-specific primers followed by direct sequencing of the PCR product for molecular identification. Overall our PCR and sequencing results confirmed pathogenic genotypes including T4 and T15 from both soil and water samples. This is our first report of Acanthamoeba isolation from both soil and water resources of Pakistan which may serve as a potential treat to human health across the country.

  9. Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea.

    Science.gov (United States)

    Visvesvara, Govinda S; Moura, Hercules; Schuster, Frederick L

    2007-06-01

    Among the many genera of free-living amoebae that exist in nature, members of only four genera have an association with human disease: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri and Sappinia diploidea. Acanthamoeba spp. and B. mandrillaris are opportunistic pathogens causing infections of the central nervous system, lungs, sinuses and skin, mostly in immunocompromised humans. Balamuthia is also associated with disease in immunocompetent children, and Acanthamoeba spp. cause a sight-threatening infection, Acanthamoeba keratitis, mostly in contact-lens wearers. Of more than 30 species of Naegleria, only one species, N. fowleri, causes an acute and fulminating meningoencephalitis in immunocompetent children and young adults. In addition to human infections, Acanthamoeba, Balamuthia and Naegleria can cause central nervous system infections in animals. Because only one human case of encephalitis caused by Sappinia diploidea is known, generalizations about the organism as an agent of disease are premature. In this review we summarize what is known of these free-living amoebae, focusing on their biology, ecology, types of disease and diagnostic methods. We also discuss the clinical profiles, mechanisms of pathogenesis, pathophysiology, immunology, antimicrobial sensitivity and molecular characteristics of these amoebae.

  10. Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran

    Science.gov (United States)

    Shokri, Azar; Sarvi, Shahabeddin; Daryani, Ahmad; Sharif, Mehdi

    2016-01-01

    Acanthamoeba, a free-living amoeba, is widely distributed in the environment, water sources, soil, dust, and air. It can cause keratitis in contact lens wearers with poor hygiene and also fatal granulomatous amebic encephalitis (GAE) in immunocompromised hosts. The aim of this study was to gain some insights into the distribution and genotypes of the potentially pathogenic species of Acanthamoeba present in water sources in north of Iran. Total 43 Acanthamoeba species were isolated from 77 water samples taken from different water sources within the Mazandaran province in Northern Iran (Sari city and suburbs). Isolates were identified based on cyst and trophozoite morphological characteristics as well genetics. PCR fragments corresponding to the small-subunit 18S rRNA gene were sequenced for 20 of 43 positive isolates. The results revealed that 83.3% of sequenced isolates belonged to the T4 genotype and the rest belonged to the T2 genotype. Our results indicated that Acanthamoeba is widely distributed in Sari city. As the incidence in Iran of amoebic keratitis has increased in recent years, the exact estimation of the prevalence of this amoeba and its predominant genotype may play a crucial role in prevention of the disease. Sari city has several rivers, seashores, and natural recreational amenities, which attract visitors during the year. This is the first report of Acanthamoeba genotypes from water sources in Sari city, Mazandaran province of Iran, and the results suggest that more attention is needed to protect the visiting population and immunocompromised individuals. PMID:27658596

  11. Physiological, morphological, and immunochemical parameters used for the characterization of clinical and environmental isolates of Acanthamoeba.

    Science.gov (United States)

    Becker-Finco, A; Costa, A O; Silva, S K; Ramada, J S; Furst, C; Stinghen, A E; De Figueiredo, B C; De Moura, J; Alvarenga, L M

    2013-03-01

    The factors that characterize Acanthamoeba strains as harmless or potentially pathogenic have not been elucidated. Analysing the in vitro and in vivo parameters of Acanthamoeba samples, including heat tolerance at temperatures close to that of the human body, cytopathic effects, and their ability to cause infections in animals, has been proposed to identify their pathogenic potential. Another promising criterion for differentiating strains is the analysis of their biochemical and immunochemical properties. In this study, a comparative evaluation between clinical and environmental Acanthamoeba isolates was performed on the basis of physiological, morphological, and immunochemical criteria. Crude antigens were used to characterize the protein profiles by electrophoresis and immunize mice to produce polyclonal and monoclonal antibodies. The antibodies were characterized using ELISA, Western blotting, and immunofluorescence techniques. The results obtained with polyclonal antibodies suggest the presence of specific proteins for each studied isolate and co-reactive immunochemical profiles among conserved components. Ten monoclonal antibody clones were obtained; mAb3 recognized 3 out of 4 samples studied. The results of this study may help standardize criteria for identifying and characterizing Acanthamoeba strains. Taken together, our results support the view that a set of features may help differentiate Acanthamoeba species and isolates.

  12. A multisystemic Acanthamoeba infection in a dog in Tenerife, Canary Islands, Spain.

    Science.gov (United States)

    Valladares, María; Reyes-Batlle, María; Mora-Peces, Inmaculada; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Dorta-Gorrín, Alexis; Comyn-Afonso, Estefanía; Martínez-Carretero, Enrique; Maciver, Sutherland K; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

    2014-10-15

    A 22-month-old male Spanish water dog was hospitalized after its physical examination revealed fever and movement difficulty. After 24h, the dog was found to have a high fever (39.5 °C) and was treated empirically with doxycycline/ciprofloxacin. At 48 h, after submission the fever rose to 41 °C and the animal presented with a stiff neck and dehydration. Peripheral blood and cerebrospinal fluid (CSF) were sampled and trophozoites with an Acanthamoeba-like morphology were observed in the CSF. PCR specific for Acanthamoeba, Naegleria fowleri and Balamuthia mandrillaris were performed and the CSF sample found positive for Acanthamoeba. Lungs, kidney, liver and spleen samples were collected post mortem. All collected organ samples were positive for Acanthamoeba by PCR, thus confirming a multisystemic infection. Water samples taken at a suspected site of infection yielded an almost identical PCR fragment to those of the clinical samples, indicating that this was probably where the infection originated. This is the first report of a fatal case of Acanthamoeba disseminated infection in a dog in Spain.

  13. Identification and typing of free-living Acanthamoeba spp. by MALDI-TOF MS Biotyper.

    Science.gov (United States)

    Del Chierico, Federica; Di Cave, David; Accardi, Cristel; Santoro, Maristella; Masotti, Andrea; D'Alfonso, Rossella; Berrilli, Federica; Urbani, Andrea; Putignani, Lorenza

    2016-11-01

    Over the years, the potential pathogenicity of Acanthamoeba for humans and animals has gained increasing attention from the scientific community. More than 24 species belong to this genus, however only some of them are causative agents of keratitis and encephalitis in humans. Due to technical difficulties in diagnosis, these infections are likely to be under-detected. The introduction of 18S rDNA amplification for the identification of Acanthamoeba has dramatically enhanced diagnosis performances, but the attestation of genotyping requires supplementary sequencing-based procedures. In this study, 15 Acanthamoeba strains were collected and grown on nutrient agar media. Each strain was genotyped by end-point PCR assay for the amplification of the 18S rDNA gene and the genotype was assigned by sequencing analysis through neighbor joining phylogenetic tree. In order to optimize standardization of the MALDI-TOF MS assay, we established the collection time point at the cystic phase. Two strains of each genotype were randomly chosen to customize the biotyper database. For all strains, 24 spectral measurements were acquired and submitted to identification and cluster analysis of spectra. The obtained results highlighted the correct identification of Acanthamoeba strains and the overlapping of spectra dendrogram clusters to the 18S genotype assignations. In conclusion, the MALDI-TOF MS Biotyper revealed the capability to identify and genotype the Acanthamoeba strains, providing a new frontier in the diagnostic identification of amaebae and in taxonomic and phylogenetic studies.

  14. Acanthamoeba genotypes T3 and T4 as causative agents of amoebic keratitis in Mexico.

    Science.gov (United States)

    Omaña-Molina, Maritza; Vanzzini-Zago, Virginia; Hernandez-Martinez, Dolores; Gonzalez-Robles, Arturo; Salazar-Villatoro, Lizbeth; Ramirez-Flores, Elizabeth; Oregon-Miranda, Eric; Lorenzo-Morales, Jacob; Martinez-Palomo, Adolfo

    2016-02-01

    Free-living amoebae (FLA) are widely distributed worldwide. Some genera included in this group act as opportunistic pathogens causing fatal encephalitis and Acanthamoeba keratitis (AK), a sight-threatening infection of the cornea associated with the use of soft contact lenses that could even end in blindness if an early diagnosis and treatment are not achieved. Furthermore, the numbers of AK cases keep rising worldwide mainly due to an increase of contact lens wearers and lack of hygiene in the maintenance of lenses and their cases. In Mexico, no cases of AK have been described so far although the isolation of other pathogenic FLA such as Naegleria fowleri and Balamuthia mandrillaris from both clinical and environmental sources has been reported. The present study reports two cases of Acanthamoeba keratitis diagnosed in two patients admitted to the Hospital "Luis Sánchez Bulnes" for Blindness Prevention in Mexico City, Mexico. Corneal scrapes and contact lenses were checked for the presence of Acanthamoeba strains in both patients. Strains were axenized after initial isolation to classify at the genotype level. After sequencing the diagnostic fragment 3 (DF3) region located on the 18S ribosomal DNA (rDNA) gene of Acanthamoeba, genotype T3 and genotype T4 were identified in clinical case 1 and 2, respectively. To our knowledge, these are the first reported cases of AK in Mexico in the literature and the first description of Acanthamoeba genotypes T3 and T4 as causative agents of amoebic infection.

  15. Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran.

    Science.gov (United States)

    Shokri, Azar; Sarvi, Shahabeddin; Daryani, Ahmad; Sharif, Mehdi

    2016-08-01

    Acanthamoeba, a free-living amoeba, is widely distributed in the environment, water sources, soil, dust, and air. It can cause keratitis in contact lens wearers with poor hygiene and also fatal granulomatous amebic encephalitis (GAE) in immunocompromised hosts. The aim of this study was to gain some insights into the distribution and genotypes of the potentially pathogenic species of Acanthamoeba present in water sources in north of Iran. Total 43 Acanthamoeba species were isolated from 77 water samples taken from different water sources within the Mazandaran province in Northern Iran (Sari city and suburbs). Isolates were identified based on cyst and trophozoite morphological characteristics as well genetics. PCR fragments corresponding to the small-subunit 18S rRNA gene were sequenced for 20 of 43 positive isolates. The results revealed that 83.3% of sequenced isolates belonged to the T4 genotype and the rest belonged to the T2 genotype. Our results indicated that Acanthamoeba is widely distributed in Sari city. As the incidence in Iran of amoebic keratitis has increased in recent years, the exact estimation of the prevalence of this amoeba and its predominant genotype may play a crucial role in prevention of the disease. Sari city has several rivers, seashores, and natural recreational amenities, which attract visitors during the year. This is the first report of Acanthamoeba genotypes from water sources in Sari city, Mazandaran province of Iran, and the results suggest that more attention is needed to protect the visiting population and immunocompromised individuals.

  16. Isolation and genotyping of acanthamoeba strains from soil sources from Jamaica, West Indies.

    Science.gov (United States)

    Todd, Cheridah D; Reyes-Batlle, María; Martín-Navarro, Carmen Ma; Dorta-Gorrín, Alexis; López-Arencibia, Atteneri; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lindo, John F; Lorenzo-Morales, Jacob

    2015-01-01

    Acanthamoeba spp. are opportunistic pathogens that are ubiquitous in nature. Many species of this genus are responsible for a fatal encephalitis and keratitis in humans and other animals. Seventy-two soil samples were collected from the parishes across Jamaica and assessed for the presence of Acanthamoeba spp. Cultivation was carried out on non-nutrient agar plates seeded with heat killed Escherichia coli. PCR and sequencing of the DF3 region were carried out in order to genotype the isolated strains of Acanthamoeba. Thermotolerance and osmotolerance assays were utilized to investigate the pathogenic potential of the Acanthamoeba isolates. Acanthamoeba spp. was isolated from 63.9% of soil samples. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T4, T5, and T11. T4 genotype was most frequently isolated. Most isolates were thermotolerant or both thermotolerant and osmotolerant, indicating that they may present the potential to cause disease in humans and other animals.

  17. Programmed cell death in Acanthamoeba castellanii Neff induced by several molecules present in olive leaf extracts.

    Science.gov (United States)

    Sifaoui, Ines; López-Arencibia, Atteneri; Martín-Navarro, Carmen Mª; Reyes-Batlle, María; Wagner, Carolina; Chiboub, Olfa; Mejri, Mondher; Valladares, Basilio; Abderrabba, Manef; Piñero, José E; Lorenzo-Morales, Jacob

    2017-01-01

    Therapy against Acanthamoeba infections such as Granulomatous Amoebic Encephalitis (GAE) and Acanthamoeba Keratitis (AK), remains as an issue to be solved due to the existence of a cyst stage which is highly resistant to most chemical and physical agents. Recently, the activity of Olive Leaf Extracts (OLE) was demonstrated against Acanthamoeba species. However, the molecules involved in this activity were not identified and/or evaluated. Therefore, the aim of this study was to evaluate the activity of the main molecules which are present in OLE and secondly to study their mechanism of action in Acanthamoeba. Among the tested molecules, the observed activities ranged from an IC50 of 6.59 in the case of apigenine to an IC50 > 100 μg/ml for other molecules. After that, elucidation of the mechanism of action of these molecules was evaluated by the detection of changes in the phosphatidylserine (PS) exposure, the permeability of the plasma membrane, the mitochondrial membrane potential and the ATP levels in the treated cells. Vanillic, syringic and ursolic acids induced the higher permeabilization of the plasma membrane. Nevertheless, the mitochondrial membrane was altered by all tested molecules which were also able to decrease the ATP levels to less than 50% in IC90 treated cells after 24 h. Therefore, all the molecules tested in this study could be considered as a future therapeutic alternative against Acanthamoeba spp. Further studies are needed in order to establish the true potential of these molecules against these emerging opportunistic pathogenic protozoa.

  18. Riboflavin and ultraviolet-A as adjuvant treatment against Acanthamoeba cysts

    Science.gov (United States)

    Lamy, Ricardo; Chan, Elliot; Good, Samuel D; Cevallos, Vicky; Porco, Travis C; Stewart, Jay M

    2015-01-01

    Background Experimental studies have shown that the standard dose of R or R+UVA as solo treatment are not able to exterminate Acanthamoeba cysts or even trophozoites. The purpose of this study is to determine whether the application of R+UVA can enhance the cysticidal effects of cationic antiseptic agents in vitro. Methods The log of either polyhexamethylene biguanide (PHMB) or chlorhexidine minimal cysticidal concentration (MCC) in solutions containing riboflavin (concentrations 0.1 %; 0.05% and 0.025 %) plus either Acanthamoeba castellanii cysts or Acanthamoeba polyphaga cysts was determined and compared in groups treated with UVA 30 mW/cm2 for 30 min and in control groups (with no exposure to UVA). A permutation test was used to determine the P-value associated with treatment. Results Regardless of the riboflavin concentration and UVA treatment condition, no trophozoites were seen in plates where the cysts were previously exposed to cationic antiseptic agents concentrations ≥ 200 µg/mL for Acanthamoeba castellanii samples and ≥ 100 µg/mL for Acanthamoeba polyphaga samples. There was no statistical evidence that R+UVA treatment was associated with MCC (P = 0.82). Conclusion R+UVA in doses up to 10 times higher than recommended for corneal crosslinking does not enhance the cysticidal effect of either polyhexamethylene biguanide or chlorhexidine in vitro. PMID:26355273

  19. Evaluation of the effectiveness of tea tree oil in treatment of Acanthamoeba infection.

    Science.gov (United States)

    Hadaś, Edward; Derda, Monika; Cholewiński, Marcin

    2017-03-01

    Eye diseases caused by amoebae from the genus Acanthamoeba are usually chronic and severe, and their treatment is prolonged and not very effective. The difficulties associated with therapy have led to attempts at finding alternative treatment methods. Particularly popular is searching for cures among drugs made of plants. However, no substances with total efficacy in treating Acanthamoeba keratitis have been identified.Results of our semi in vivo studies of tea tree oil simulating eyeball infection demonstrated 100% effectiveness in the case of both trophozoites and cysts of amoebae from the genus Acanthamoeba. The action of tea tree oil indicates that this is the first substance with a potential ability to quickly and effectively remove the amoebae from the eye. Tea tree oil has the ability to penetrate tissues, which allows it to destroy amoebae in both the shallow and deep layers of the cornea. The present research into the use of tea tree oil in the therapy of Acanthamoeba infection is the first study of this type in parasitology. It offers tremendous potential for effective treatment of Acanthamoeba keratitis and other diseases caused by these protozoa.

  20. Chloroquine has a cytotoxic effect on Acanthamoeba encystation through modulation of autophagy.

    Science.gov (United States)

    Jha, Bijay Kumar; Jung, Hui-Jung; Seo, Incheol; Kim, Hyun Ah; Suh, Seong-Il; Suh, Min-Ho; Baek, Won-Ki

    2014-10-01

    Encystation of Acanthamoeba castellanii is associated with resistance to chemotherapeutic agents. Blocking the encystation process could potentiate the efficacy of chemotherapeutic agents and biocides. During encystation, autophagy is highly stimulated and required for proper encystation of Acanthamoeba. In this study, the cytotoxic effect of chloroquine, a well-known autophagy-inhibitory drug, was tested in A. castellanii. Chloroquine was able to selectively reduce cell survival during the encystation of A. castellanii. However, A. castellanii trophozoites and mature cysts were resistant to chloroquine. Chloroquine treatment led to an increase in the number and size of lysosomes in encysting cells. Moreover, chloroquine inhibited the degradation of long-lived proteins in the encysting cells. Decreased autophagic flux, indicated by an increased number of lysosomes and decreased degradation of long-lived proteins, may be the mechanism by which cell death is induced by chloroquine in encysting Acanthamoeba. These results suggest a potential novel therapeutic application of chloroquine as an anti-Acanthamoeba drug. Our findings also suggest that targeting autophagy could be a therapeutic strategy against Acanthamoeba infection.

  1. Detection and quantification of human adenovirus genomes in Acanthamoeba isolated from swimming pools

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    RODRIGO STAGGEMEIER

    2016-01-01

    Full Text Available ABSTRACT Acanthamoeba is the most common free-living environmental amoeba, it may serve as an important vehicle for various microorganisms living in the same environment, such as viruses, being pathogenic to humans. This study aimed to detect and quantify human adenoviruses (HAdV in Acanthamoebas isolated from water samples collected from swimming pools in the city of Porto Alegre, Southern Brazil. Free-living amoebae of the genus Acanthamoeba were isolated from water samples, and isolates (n=16 were used to investigate the occurrence of HAdVs. HAdV detection was performed by quantitative real-time polymerase chain reaction (qPCR. HAdVs were detected in 62.5% (10/16 of Acanthamoeba isolates, ranging from 3.24x103 to 5.14x105 DNA copies per milliliter of isolate. HAdV viral loads found in this study are not negligible, especially because HAdV infections are associated with several human diseases, including gastroenteritis, respiratory distress, and ocular diseases. These findings reinforce the concept that Acanthamoeba may act as a reservoir and promote HAdV transmission through water.

  2. Uso del cefadroxilo oral en las neumopatías inflamatorias adquiridas en la comunidad

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    Juan José Pisonero Socías

    1999-12-01

    Full Text Available Se realizó un estudio del uso del cefadroxilo (cefalosporina oral de 1ra. generación en 100 pacientes con diagnóstico de sespsis respiratoria baja extrahospitalaria, donde se utilizó un esquema de dosis de 500 mg de cada 12 horas durante 7 a 10 días. A 60 pacientes se les administró 1 g por 7 días, de los cuales curaron 49 para el 81 %; de los 36 pacientes a los que se les administró 1 g por 10 días curaron 28, lo que representa el 77 % y de los 4 restantes que recibieron una dosis de 2 g por 10 días curaron 2 para el 50 %. Se efectuó además un análisis comparativo de los costos en tratamientos con penicilina cristalina, cefazolina, ceftriaxona y aminoglucósidos en la misma afección y se logró una disminución en el costo hospitalario y el consumo de antibióticosA study on the use of cefadroxil (1st generation oral cephalosporins was conducted among 100 patients with diagnosis of extrahospital lower respiratory sepsis. A dosage schedule of 500 mg every 12 hours during 7 or 10 days was used. 49 (81 % of the 60 patients that were administered 1 g during 7 days healed, whereas 28 (77 % of those who received 1 g for 10 days cured, too. 2(50 % of the other 4 that were given doses of 2 g for 10 days recovered from the disease. A comparative analysis of the costs generated by the use of crystalline penicillin, ceftriaxone, cefazolin and aminoglycosides to treat the same affection was made, and a reduction of hospital cost and antibiotic consumption was attained

  3. La peroxidación lipídica en la etiopatogenia de la enfermedad periodontal inflamatoria

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    Bárbara E García Triana

    Full Text Available Las especies reactivas del oxígeno están implicadas en la etiopatogenia de la inflamación. Durante la activación de los leucocitos, se liberan grandes cantidades de estas especies, cuya función es la eliminación de los agentes patógenos. Si las defensas antioxidantes de los tejidos no funcionan eficientemente, son inducidas reacciones radicálicas que afectan a las biomoléculas. El ataque a los lípidos de la membrana celular provoca su peroxidación, con la consiguiente formación de nuevas especies radicálicas y metabolitos tóxicos. La presencia de un fuerte infiltrado inflamatorio en los tejidos periodontales, durante la enfermedad periodontal inflamatoria, ha sugerido la posible participación de las especies reactivas del oxígeno en la etiopatogenia de esta enfermedad. Se supone que el ataque de estas sustancias a los tejidos periodontales con deficiente defensa antioxidante, provoca la aparición de la peroxidación lipídica, que puede conducir a la lisis celular y la activación de proteasas.

  4. Bases de la respuesta inflamatoria en la forma respiratoria del PRRS

    OpenAIRE

    Barranco Cabezudo, Inmaculada; Quereda, J.J.; García-Nicolás, O.; Ramis, G.; Pallarés, F.J.; Rodríguez-Gómez, I. M.; Gómez-Laguna, J.; Carrasco Otero, Librado

    2011-01-01

    El Síndrome Reproductivo y Respiratorio Porcino (PRRS) es una enfermedad de distribución mundial que causa graves pérdidas económicas al sector porcino. Este virus no sólo es importante como agente causal del PRRS sino también por su participación en el desarrollo del Complejo Respiratorio Porcino. Su interacción con las defensas pulmonares, la alteración de la respuesta inmune y su persistencia en los órganos linfoides conlleva a que los cerdos tengan dificultades para luchar contra la enfer...

  5. Monografía sobre el intestino grueso : enfermedades inflamatorias intestinales

    OpenAIRE

    López Mora, Gema

    2014-01-01

    El intestino grueso es un órgano con forma de tubo, de aproximadamente un metro y medio de largo, que forma parte del sistema digestivo. Consta de varias partes llamadas ciego (con su apéndice vermiforme); colon, en donde ponemos diferenciar colon ascendente, transverso, descendente y sigmoide; recto y ano. Sus paredes están cubiertas por un epitelio simple cilíndrico, que no presenta pliegues ni vellosidades y, que forma numerosas glándulas mucosas tubulares llamadas criptas de Lierberkühn, ...

  6. Lethal effects of Helianthemum lippii (L.) on Acanthamoeba castellanii cysts in vitro.

    Science.gov (United States)

    Badria, F A; Hetta, M H; Sarhan, Rania M; Ezz El-Din, M H

    2014-06-01

    Acanthamoeba spp. commonly cause Acanthamoeba keratitis which is typically associated with the wear of contact lenses. Therefore, finding an economic, efficient, and safe therapy of natural origin is of outmost importance. This study examined the in vitro lethal potential of ethyl acetate and methanol extracts of Helianthemum lippii (L.) (sun roses) against Acanthamoeba castellanii cysts isolated from patients with amoebic keratitis. Both extracts proved to be potent as regard to their lethal effects on A. castellanii cysts with comparable results to chlorhexidine. The ethyl acetate was more promising with cumulative lethality. It showed a highly significant lethal percentage along the duration of treatment. The analysis of the more potent ethyl acetate extract revealed the presence of 2.96 mg/100 g of total phenolics, 0.289 mg/100 ml of total flavonoids and 37 mg/100 mg of total tannins which highlighted their phytomedicinal role.

  7. Acanthamoeba keratitis presenting as dendritic keratitis in a soft contact lens wearer.

    Science.gov (United States)

    Yeung, Edward Yip; Huang, Samuel Chao-Ming; Tsai, Ray Jui-Fang

    2002-03-01

    Acanthamoeba keratitis is a rare cause of corneal infection in Taiwan, which can result in devastating visual outcomes. A 37-year-old woman, who wore soft contact lenses, suffered from severe pain in her left eye. Biomicroscopy revealed dendritic keratitis, radial keratoneuritis, and fine keratic precipitates on her cornea. Culture, using non-nutrient agar plate seeded with Escherichia coli, resulted in heavy growth of Acanthamoeba. The inpatient treatment, including topical neomycin-polymyxin B and metronidazole (0.5%) eyedrops, oral ketoconazole, and then oral prednisolone, successfully controlled the corneal infection. The best-corrected visual acuity was 0.9 without any evidence of recurrence of infection after 21 months of follow up. Acanthamoeba keratitis can present as dendritic keratitis, which mimics herpes simplex infection, thus, delays appropriate treatment. Early diagnosis and judicious treatment are essential for restoring the vision and avoiding the subsequent need of penetrating keratoplasty.

  8. Isolation and identification of Acanthamoeba species from natural water sources in the northeastern part of Thailand.

    Science.gov (United States)

    Thammaratana, Thani; Laummaunwai, Porntip; Boonmars, Thidarut

    2016-04-01

    Acanthamoeba are found in the environment, particularly in water, all over the world. The genus is currently classified into 20 different genotypes, T1-T20. In this study, 63 natural water samples from 11 provinces in northeast Thailand were collected and cultured on non-nutrient agar plates. Positive samples by culture were subsequently analyzed by molecular methods. The identification of Acanthamoeba was based on morphological features and molecular techniques using PCR and DNA sequencing. The results showed that 10 samples out of 63 were positive (15.9 %). Phylogenetic analysis revealed that seven samples were T4, one sample was similar to T3, and the other two samples were similar to T5. This is the first report demonstrating the contamination of Acanthamoeba species in natural water sources in northeast Thailand.

  9. Axenic Cultivation and Pathogenic Assays of Acanthamoeba Strains Using Physical Parameters

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    M Niyyati

    2013-06-01

    Full Text Available Background: The main goal of the present study was to set up an axenic cultivation of Acanthamoeba and assess the pathogenic ability of T4 genotypes from different clinical and environmental strains of Acanthamoeba using two physical assays.Methods: Sixteen Acanthamoeba isolates including 10 environmental and 6 clinical strains were cul­tured axenically. Axenic cultivation was performed using Proteosepepton, yeast extract and glucose medium and TY-I-S33culture. Pathogenic survey was done using osmotolerance and thermotoler­ance assay. Briefly, differentosmolarity (0.5 M and 1 M of non-nutrient agar plates were performed. One hundred fiftyµl of axenic culture were collected and were inoculated in 1% agar medium. For thermotolerance assay 150 µl of amoebas from axenic culture were divided into fresh culture me­diums. Cultures were incubated at 37oC and 42 oC. All plates were monitored for 24 h, 48 h and 72 h.Results: Overall, 16 strains of Acanthamoeba isolates previously genotyped as T4 were cultivated axeni­cally after several months. Thermotolerance and osmotolerance assay revealed that all of clinical strains, soil and animal feces strains were highly pathogenic isolates. Two dust and water strains did not grow at high temperature (42 oC and osmolarity (1.5 M and thus they were classified as weak pathogens.Conclusion: Most of T4 genotypes are highly pathogenic organisms. This is an important finding since Acanthamoeba belonging to T4 type is the predominate genotype in environmental and clinical samples. The presence of highly pathogenic Acanthamoeba may pose a risk within susceptible people.

  10. Pathogenic strains of Acanthamoeba are recognized by TLR4 and initiated inflammatory responses in the cornea.

    Science.gov (United States)

    Alizadeh, Hassan; Tripathi, Trivendra; Abdi, Mahshid; Smith, Ashley Dawn

    2014-01-01

    Free-living amoebae of the Acanthamoeba species are the causative agent of Acanthamoeba keratitis (AK), a sight-threatening corneal infection that causes severe pain and a characteristic ring-shaped corneal infiltrate. Innate immune responses play an important role in resistance against AK. The aim of this study is to determine if Toll-like receptors (TLRs) on corneal epithelial cells are activated by Acanthamoeba, leading to initiation of inflammatory responses in the cornea. Human corneal epithelial (HCE) cells constitutively expressed TLR1, TLR2, TLR3, TLR4, and TLR9 mRNA, and A. castellanii upregulated TLR4 transcription. Expression of TLR1, TLR2, TLR3, and TLR9 was unchanged when HCE cells were exposed to A. castellanii. IL-8 mRNA expression was upregulated in HCE cells exposed to A. castellanii. A. castellanii and lipopolysaccharide (LPS) induced significant IL-8 production by HCE cells as measured by ELISA. The percentage of total cells positive for TLR4 was higher in A. castellanii stimulated HCE cells compared to unstimulated HCE cells. A. castellanii induced upregulation of IL-8 in TLR4 expressing human embryonic kidney (HEK)-293 cells, but not TLR3 expressing HEK-293 cells. TLR4 neutralizing antibody inhibited A. castellanii-induced IL-8 by HCE and HEK-293 cells. Clinical strains but not soil strains of Acanthamoeba activated TLR4 expression in Chinese hamster corneas in vivo and in vitro. Clinical isolates but not soil isolates of Acanthamoeba induced significant (PAcanthamoeba activate TLR4 and induce production of CXCL2 in the Chinese hamster model of AK. TLR4 may be a potential target in the development of novel treatment strategies in Acanthamoeba and other microbial infections that activate TLR4 in corneal cells.

  11. Evaluation of the immunodiagnostic potential of a recombinant surface protein domain from Acanthamoeba castellanii.

    Science.gov (United States)

    Sánchez, Alemao G Carpinteyro; Virginio, Veridiana Gomes; Maschio, Vinicius José; Ferreira, Henrique Bunselmeyer; Rott, Marilise Brittes

    2016-10-01

    Acanthamoeba spp. are free-living protists widely distributed in environment, able to cause keratitis, encephalitis and skin lesions in humans and animals. Acanthamoeba spp. exist in two forms: an infective trophozoite and a dormant cyst. Several factors contribute to the pathogenesis of Acanthamoeba spp. The parasite adhesion to the host cell is the primary step for infection and is mediated by a mannose binding-protein, expressed in the surface and considered the main pathogenicity factor in Acanthamoeba spp. So far, there was no evidence of another surface protein of Acanthamoeba spp. relevant for host invasion or infection by these organisms. The aims of this study were to identify and characterize an Acanthamoeba castellanii surface protein and to evaluate its diagnostic potential. In silico predictions of surface proteins allowed to identify the A. castellanii calreticulin as a possible surface antigen. The coding sequence of a predicted extracellular domain of A. castellanii calreticulin was cloned by in vivo homologous recombination and the recombinant polypeptide (AcCRT29-130) was produced. Its immunodiagnostic potential was assessed in a recombinant antigen-based ELISA with sera from experimentally infected rats that developed keratitis and encephalitis, and sera from patients with encephalitis. The AcCRT29-130 was significantly more recognized by sera from encephalitis infected rats in comparison with the non-infected controls. Human sera from encephalitis patients, however presented no significant response. These results showed the AcCRT29-130 potential for A. castellanii infection immunodiagnosis in animals, with further studies being required for assessment of its use for human infections.

  12. A rapid and reliable method for the clonal isolation of Acanthamoeba from environmental samples

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    Janice Zanella

    2012-02-01

    Full Text Available Acanthamoeba are abundant in a wide range of environments, and some species are responsible for cutaneous infections, keratitis, and granulomatous amoebic encephalitis (GAE. The conventional detection and isolation of amoeba from clinical and environmental samples involves sampling and culture on non-nutrient Ágar medium. Although efficient, this system requires several transfers in order to eliminate contaminants, and is not appropriate for the isolation of individual amoeba from samples with a biodiverse community. In this study we propose an alternative method for the isolation of monocystic clones of Acanthamoeba. The propose method involves sampling, enrichment, encystment induction, and direct cysts micromanipulation and culture on Ágar plates.

  13. Novel Acanthamoeba 18S rRNA gene sequence type from an environmental isolate.

    Science.gov (United States)

    Magnet, A; Henriques-Gil, N; Galván-Diaz, A L; Izquiedo, F; Fenoy, S; del Aguila, C

    2014-08-01

    The free-living amoebae, Acanthamoeba, can act as opportunistic parasites on a wide range of vertebrates and are becoming a serious threat to human health due to the resistance of their cysts to harsh environmental conditions, disinfectants, some water treatment practices, and their ubiquitous distribution. Subgenus classification based on morphology is being replaced by a classification based on the sequences of the 18S rRNA gene with a total of 18 different genotypes (T1-T18). A new environmental strain of Acanthamoeba isolated from a waste water treatment plant is presented in this study as a candidate for the description of the novel genotype T19 after phylogenetic analysis.

  14. Killing of diverse eye pathogens (Acanthamoeba spp., Fusarium solani, and Chlamydia trachomatis) with alcohols

    Science.gov (United States)

    2017-01-01

    Background Blindness is caused by eye pathogens that include a free-living protist (Acanthamoeba castellanii, A. byersi, and/or other Acanthamoeba spp.), a fungus (Fusarium solani), and a bacterium (Chlamydia trachomatis). Hand-eye contact is likely a contributor to the spread of these pathogens, and so hand washing with soap and water or alcohol–based hand sanitizers (when water is not available) might reduce their transmission. Recently we showed that ethanol and isopropanol in concentrations present in hand sanitizers kill walled cysts of Giardia and Entamoeba, causes of diarrhea and dysentery, respectively. The goal here was to determine whether these alcohols might kill infectious forms of representative eye pathogens (trophozoites and cysts of Acanthamoeba, conidia of F. solani, or elementary bodies of C. trachomatis). Methodology/Principal findings We found that treatment with 63% ethanol or 63% isopropanol kills >99% of Acanthamoeba trophozoites after 30 sec exposure, as shown by labeling with propidium iodide (PI) and failure to grow in culture. In contrast, Acanthamoeba cysts, which contain cellulose fibers in their wall, are relatively more resistant to these alcohols, particularly isopropanol. Depending upon the strain tested, 80 to 99% of Acanthamoeba cysts were killed by 63% ethanol after 2 min and 95 to 99% were killed by 80% ethanol after 30 sec, as shown by PI labeling and reduced rates of excystation in vitro. Both ethanol and isopropanol (63% for 30 sec) kill >99% of F. solani conidia, which have a wall of chitin and glucan fibrils, as demonstrated by PI labeling and colony counts on nutrient agar plates. Both ethanol and isopropanol (63% for 60 sec) inactivate 96 to 99% of elementary bodies of C. trachomatis, which have a wall of lipopolysaccharide but lack peptidoglycan, as measured by quantitative cultures to calculate inclusion forming units. Conclusions/Significance In summary, alcohols kill infectious forms of Acanthamoeba, F. solani, and

  15. Propylene glycol and contact-lens solutions containing this diol induce pseudocyst formation in acanthamoebae.

    Science.gov (United States)

    Kliescikova, Jarmila; Kulda, Jaroslav; Nohynkova, Eva

    2011-01-01

    Propylene glycol used as an ophthalmic demulcent in certain contact-lens care systems has been included recently among factors responsible for increasing Acanthamoeba keratitis. In this study, we provide evidence that propylene glycol as well as examined contact-lens solutions containing it induce rapid differentiation of acanthamoebae into pseudocysts. The partial resistance of the pseudocysts and their reversibility to viable trophozoites even after 24-h exposure to the contact-lens solutions indicate a potential risk of infection to contact-lens users.

  16. Killing of diverse eye pathogens (Acanthamoeba spp., Fusarium solani, and Chlamydia trachomatis with alcohols.

    Directory of Open Access Journals (Sweden)

    Yousuf Aqeel

    2017-02-01

    Full Text Available Blindness is caused by eye pathogens that include a free-living protist (Acanthamoeba castellanii, A. byersi, and/or other Acanthamoeba spp., a fungus (Fusarium solani, and a bacterium (Chlamydia trachomatis. Hand-eye contact is likely a contributor to the spread of these pathogens, and so hand washing with soap and water or alcohol-based hand sanitizers (when water is not available might reduce their transmission. Recently we showed that ethanol and isopropanol in concentrations present in hand sanitizers kill walled cysts of Giardia and Entamoeba, causes of diarrhea and dysentery, respectively. The goal here was to determine whether these alcohols might kill infectious forms of representative eye pathogens (trophozoites and cysts of Acanthamoeba, conidia of F. solani, or elementary bodies of C. trachomatis.We found that treatment with 63% ethanol or 63% isopropanol kills >99% of Acanthamoeba trophozoites after 30 sec exposure, as shown by labeling with propidium iodide (PI and failure to grow in culture. In contrast, Acanthamoeba cysts, which contain cellulose fibers in their wall, are relatively more resistant to these alcohols, particularly isopropanol. Depending upon the strain tested, 80 to 99% of Acanthamoeba cysts were killed by 63% ethanol after 2 min and 95 to 99% were killed by 80% ethanol after 30 sec, as shown by PI labeling and reduced rates of excystation in vitro. Both ethanol and isopropanol (63% for 30 sec kill >99% of F. solani conidia, which have a wall of chitin and glucan fibrils, as demonstrated by PI labeling and colony counts on nutrient agar plates. Both ethanol and isopropanol (63% for 60 sec inactivate 96 to 99% of elementary bodies of C. trachomatis, which have a wall of lipopolysaccharide but lack peptidoglycan, as measured by quantitative cultures to calculate inclusion forming units.In summary, alcohols kill infectious forms of Acanthamoeba, F. solani, and C. trachomatis, although longer times and higher ethanol

  17. Killing of diverse eye pathogens (Acanthamoeba spp., Fusarium solani, and Chlamydia trachomatis) with alcohols.

    Science.gov (United States)

    Aqeel, Yousuf; Rodriguez, Raquel; Chatterjee, Aparajita; Ingalls, Robin R; Samuelson, John

    2017-02-01

    Blindness is caused by eye pathogens that include a free-living protist (Acanthamoeba castellanii, A. byersi, and/or other Acanthamoeba spp.), a fungus (Fusarium solani), and a bacterium (Chlamydia trachomatis). Hand-eye contact is likely a contributor to the spread of these pathogens, and so hand washing with soap and water or alcohol-based hand sanitizers (when water is not available) might reduce their transmission. Recently we showed that ethanol and isopropanol in concentrations present in hand sanitizers kill walled cysts of Giardia and Entamoeba, causes of diarrhea and dysentery, respectively. The goal here was to determine whether these alcohols might kill infectious forms of representative eye pathogens (trophozoites and cysts of Acanthamoeba, conidia of F. solani, or elementary bodies of C. trachomatis). We found that treatment with 63% ethanol or 63% isopropanol kills >99% of Acanthamoeba trophozoites after 30 sec exposure, as shown by labeling with propidium iodide (PI) and failure to grow in culture. In contrast, Acanthamoeba cysts, which contain cellulose fibers in their wall, are relatively more resistant to these alcohols, particularly isopropanol. Depending upon the strain tested, 80 to 99% of Acanthamoeba cysts were killed by 63% ethanol after 2 min and 95 to 99% were killed by 80% ethanol after 30 sec, as shown by PI labeling and reduced rates of excystation in vitro. Both ethanol and isopropanol (63% for 30 sec) kill >99% of F. solani conidia, which have a wall of chitin and glucan fibrils, as demonstrated by PI labeling and colony counts on nutrient agar plates. Both ethanol and isopropanol (63% for 60 sec) inactivate 96 to 99% of elementary bodies of C. trachomatis, which have a wall of lipopolysaccharide but lack peptidoglycan, as measured by quantitative cultures to calculate inclusion forming units. In summary, alcohols kill infectious forms of Acanthamoeba, F. solani, and C. trachomatis, although longer times and higher ethanol

  18. External NAD(P)H dehydrogenases in Acanthamoeba castellanii mitochondria.

    Science.gov (United States)

    Antos-Krzeminska, Nina; Jarmuszkiewicz, Wieslawa

    2014-09-01

    The mitochondrial respiratory chain of plants and some fungi contains multiple rotenone-insensitive NAD(P)H dehydrogenases, of which at least two are located on the outer surface of the inner membrane (i.e., external NADH and external NADPH dehydrogenases). Annotated sequences of the putative alternative NAD(P)H dehydrogenases of the protozoan Acanthamoeba castellanii demonstrated similarity to plant and fungal sequences. We also studied activity of these dehydrogenases in isolated A. castellanii mitochondria. External NADPH oxidation was observed for the first time in protist mitochondria. The coupling parameters were similar for external NADH oxidation and external NADPH oxidation, indicating similar efficiencies of ATP synthesis. Both external NADH oxidation and external NADPH oxidation had an optimal pH of 6.8 independent of relevant ubiquinol-oxidizing pathways, the cytochrome pathway or a GMP-stimulated alternative oxidase. The maximal oxidizing activity with external NADH was almost double that with external NADPH. However, a lower Michaelis constant (K(M)) value for external NADPH oxidation was observed compared to that for external NADH oxidation. Stimulation by Ca(2+) was approximately 10 times higher for external NADPH oxidation, while NADH dehydrogenase(s) appeared to be slightly dependent on Ca(2+). Our results indicate that external NAD(P)H dehydrogenases similar to those in plant and fungal mitochondria function in mitochondria of A. castellanii.

  19. Shiga toxins decrease enterohaemorrhagic Escherichia coli survival within Acanthamoeba castellanii.

    Science.gov (United States)

    Chekabab, Samuel M; Daigle, France; Charette, Steve J; Dozois, Charles M; Harel, Josée

    2013-07-01

    Enterohaemorrhagic Escherichia coli (EHEC) are zoonotic pathogens transmitted to humans through contaminated water or bovine products. One of the strategies used by pathogenic bacteria to survive in aquatic environments is using free-living amoebae as hosts. Acanthamoeba castellanii is an amoeba known to host several waterborne pathogens. This study investigates the survival of EHEC with A. castellanii, which could contribute to its spread and transmission to humans. We used a gentamicin protection assay as well as fluorescence and electron microscopy to monitor the intra-amoebae survival of EHEC O157:H7 over 24 h. The results showed that EHEC were able to survive within A. castellanii and that this survival was reduced by Shiga toxins (Stx) produced by EHEC. A toxic effect mediated by Stx was demonstrated by amoebae mortality and LDH release during co-culture of EHEC and amoeba. This work describes the ability of EHEC to survive within A. castellanii, and this host-pathogen interaction is partially controlled by the Stx. Thus, this ubiquitous amoeba could represent an environmental niche for EHEC survival and transmission.

  20. The interaction of Acanthamoeba castellanii cysts with macrophages and neutrophils.

    Science.gov (United States)

    Hurt, Michael; Proy, Vincent; Niederkorn, Jerry Y; Alizadeh, Hassan

    2003-06-01

    Acanthamoeba castellanii, a free-living amoeba, causes a sight-threatening form of keratitis. Even after extensive therapies, corneal damage can be severe, often requiring corneal transplantation to restore vision. However, A. castellanii cysts are not eliminated from the conjunctiva and stroma of humans and can excyst, resulting in infection of the corneal transplant. The aim of this study was to determine whether elements of the innate immune apparatus, neutrophils and macrophages, were capable of detecting and eliminating A. castellanii cysts and to examine the mechanism by which they kill the cysts. Results show that neither innate immune cell is attracted chemotactically to intact cysts, yet both were attracted to lysed cysts. Both macrophages and neutrophils were capable of killing significant numbers of cysts, yet neutrophils were 3-fold more efficient than macrophages. Activation of macrophages with lipopolysaccharide and interferon-gamma did not increase their cytolytic ability. Conditioned medium isolated from macrophages did not lyse the cysts; however, prevention of phagocytosis by cytochalasin D inhibited 100% of macrophage-mediated killing of the cysts. Conditioned medium from neutrophils did kill significant numbers of the cysts, and this killing was blocked by quercetin, a potent inhibitor of myeloperoxidase (MPO). These results indicate that neither macrophages nor neutrophils are chemoattracted to intact cysts, yet both are capable of killing the cysts. Macrophages killed the cysts by phagocytosis, whereas neutrophils killed cysts through the secretion of MPO.

  1. Acanthamoeba polyphaga-enhanced growth of Mycobacterium smegmatis.

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    Otmane Lamrabet

    Full Text Available BACKGROUND: Mycobacterium smegmatis is a rapidly-growing mycobacterium causing rare opportunistic infections in human patients. It is present in soil and water environments where free-living amoeba also reside, but data regarding M. smegmatis-amoeba relationships have been contradictory from mycobacteria destruction to mycobacteria survival. METHODOLOGY/PRINCIPAL FINDINGS: Using optic and electron microscopy and culture-based microbial enumeration we investigated the ability of M. smegmatis mc(2 155, M. smegmatis ATCC 19420(T and M. smegmatis ATCC 27204 organisms to survive into Acanthamoeba polyphaga trophozoites and cysts. We observed that M. smegmatis mycobacteria penetrated and survived in A. polyphaga trophozoites over five-day co-culture resulting in amoeba lysis and the release of viable M. smegmatis mycobacteria without amoebal cyst formation. We further observed that amoeba-co-culture, and lysed amoeba and supernatant and pellet, significantly increased five-day growth of the three tested M. smegmatis strains, including a four-fold increase in intra-amoebal growth. CONCLUSIONS/SIGNIFICANCE: Amoebal co-culture increases the growth of M. smegmatis resulting in amoeba killing by replicating M. smegmatis mycobacteria. This amoeba-M. smegmatis co-culture system illustrates an unusual paradigm in the mycobacteria-amoeba interactions as mycobacteria have been mainly regarded as amoeba-resistant organisms. Using these model organisms, this co-culture system could be used as a simple and rapid model to probe mycobacterial factors implicated in the intracellular growth of mycobacteria.

  2. Internalization of Mycobacterium shottsii and Mycobacterium pseudoshottsii by Acanthamoeba polyphaga.

    Science.gov (United States)

    Gupta, Tuhina; Fine-Coulson, Kari; Karls, Russell; Gauthier, David; Quinn, Frederick

    2013-08-01

    Amoebae serve as environmental hosts to a variety of mycobacteria, including Mycobacterium avium and Mycobacterium marinum. Mycobacterium shottsii and Mycobacterium pseudoshottsii are waterborne species isolated from the spleens and dermal lesions of striped bass (Morone saxatilis) from the Chesapeake Bay. The optimal growth temperature for these fish isolates is 25 °C. In the present study, amoebae were examined as a potential environmental reservoir for these fish pathogens. Several studies demonstrated that M. avium bacilli replicate within the trophozoite stage and reside in large numbers within the cytosol of the cyst of the free-living amoeba Acanthamoeba polyphaga. Results from the present study showed that M. shottsii, M. pseudoshottsii, and M. marinum bacilli were internalized by A. polyphaga trophozoites within 6 h but that intracellular viability decreased by 2 to 3 logs over 10 days. While an average of 25 M. marinum bacilli were identified by electron microscopy in the cytosol of the cyst, <5 M. pseudoshottsii and no M. shottsii bacilli were observed in this location. All Mycobacterium species examined remained viable but did not replicate after encystment and subsequent 48 h incubation in 4% HCl. This concentration of HCl will kill mycobacteria but will not enter amoebal cysts. Bacterial viability studies within stages of the amoeba life cycle indicate fewer M. shottsii and M. pseudoshottsii bacilli within the trophozoite and cyst stages relative to M. marinum.

  3. Gene and genome duplication in Acanthamoeba polyphaga Mimivirus.

    Science.gov (United States)

    Suhre, Karsten

    2005-11-01

    Gene duplication is key to molecular evolution in all three domains of life and may be the first step in the emergence of new gene function. It is a well-recognized feature in large DNA viruses but has not been studied extensively in the largest known virus to date, the recently discovered Acanthamoeba polyphaga Mimivirus. Here, I present a systematic analysis of gene and genome duplication events in the mimivirus genome. I found that one-third of the mimivirus genes are related to at least one other gene in the mimivirus genome, either through a large segmental genome duplication event that occurred in the more remote past or through more recent gene duplication events, which often occur in tandem. This shows that gene and genome duplication played a major role in shaping the mimivirus genome. Using multiple alignments, together with remote-homology detection methods based on Hidden Markov Model comparison, I assign putative functions to some of the paralogous gene families. I suggest that a large part of the duplicated mimivirus gene families are likely to interfere with important host cell processes, such as transcription control, protein degradation, and cell regulatory processes. My findings support the view that large DNA viruses are complex evolving organisms, possibly deeply rooted within the tree of life, and oppose the paradigm that viral evolution is dominated by lateral gene acquisition, at least in regard to large DNA viruses.

  4. Listeria monocytogenes does not survive ingestion by Acanthamoeba polyphaga.

    Science.gov (United States)

    Akya, Alisha; Pointon, Andrew; Thomas, Connor

    2010-03-01

    Listeria monocytogenes is a ubiquitous bacterium capable of infecting humans, particularly pregnant women and immunocompromised individuals. Although the intracellular invasion and pathogenesis of listeriosis in mammalian tissues has been well studied, little is known about the ecology of L. monocytogenes , and in particular the environmental reservoir for this bacterium has not been identified. This study used short-term co-culture at 15, 22 and 37 degrees C to examine the interaction of L. monocytogenes strains with Acanthamoeba polyphaga ACO12. Survival of L. monocytogenes cells phagocytosed by monolayers of trophozoites was assessed by culture techniques and microscopy. A. polyphaga trophozoites eliminated bacterial cells within a few hours post-phagocytosis, irrespective of the incubation temperature used. Wild-type L. monocytogenes and a phenotypic listeriolysin O mutant were unable to either multiply or survive within trophozoites. By contrast, Salmonella enterica serovar Typhimurium C5 cells used as controls were able to survive and multiply within A. polyphaga trophozoites. The data presented indicate that A. polyphaga ACO12 is unlikely to harbour L. monocytogenes, or act as an environmental reservoir for this bacterium.

  5. Legionella pneumophila prevents proliferation of its natural host Acanthamoeba castellanii

    Science.gov (United States)

    Mengue, Luce; Régnacq, Matthieu; Aucher, Willy; Portier, Emilie; Héchard, Yann; Samba-Louaka, Ascel

    2016-01-01

    Legionella pneumophila is a ubiquitous, pathogenic, Gram-negative bacterium responsible for legionellosis. Like many other amoeba-resistant microorganisms, L. pneumophila resists host clearance and multiplies inside the cell. Through its Dot/Icm type IV secretion system, the bacterium injects more than three hundred effectors that modulate host cell physiology in order to promote its own intracellular replication. Here we report that L. pneumophila prevents proliferation of its natural host Acanthamoeba castellanii. Infected amoebae could not undergo DNA replication and no cell division was observed. The Dot/Icm secretion system was necessary for L. pneumophila to prevent the eukaryotic proliferation. The absence of proliferation was associated with altered amoebal morphology and with a decrease of mRNA transcript levels of CDC2b, a putative regulator of the A. castellanii cell cycle. Complementation of CDC28-deficient Saccharomyces cerevisiae by the CDC2b cDNA was sufficient to restore proliferation of CDC28-deficient S. cerevisiae and suggests for the first time that CDC2b from A. castellanii could be functional and a bona fide cyclin-dependent kinase. Hence, our results reveal that L. pneumophila impairs proliferation of A. castellanii and this effect could involve the cell cycle protein CDC2b. PMID:27805070

  6. Staphylococcus aureus exhibit similarities in their interactions with Acanthamoeba and ThP1 macrophage-like cells.

    Science.gov (United States)

    Cardas, Mihaela; Khan, Naveed Ahmed; Alsam, Selwa

    2012-12-01

    Staphylococcus aureus is a leading cause of nosocomial infections. Haematogenous spread is a pre-requisite but it is not clear how S. aureus survive the onslaught of macrophages. Acanthamoeba is a protozoan pathogen that is remarkably similar to macrophages, particularly in their cellular structure (morphological and ultra-structural features), molecular motility, biochemical physiology, ability to capture prey by phagocytosis and interactions with microbial pathogens. Thus, we hypothesize that S. aureus exhibit similarities in their interactions with Acanthamoeba and ThP1 macrophage-like cells. Here, we studied interactions of methicillin-sensitive S. aureus (MSSA), methicillin-resistant S. aureus (MRSA) and Staphylococcus epidermidis (SE) with Acanthamoeba castellanii belonging to the T4 genotype and macrophage-like cells (ThP1). The findings revealed that both MRSA and MSSA exhibited similarities in their binding/association and invasion of A. castellanii and ThP1 cells. Long-term incubation showed that MRSA and MSSA can survive intracellularly of both Acanthamoeba and ThP1 cells. Overall, these findings suggest that Acanthamoeba exhibit similar characteristics with ThP1 macrophage-like cells in their interaction with MRSA and MSSA. Additionally it was shown that bacteria survive inside Acanthamoeba during the encystment process as evidenced by bacterial recovery from mature cysts. Given that Acanthamoeba cysts are airborne, these findings suggest that cysts may act as "Trojan horse" to help spread MRSA to susceptible hosts.

  7. Adherence to treatment in inflammatory bowel disease Adhesión al tratamiento en la enfermedad inflamatoria intestinal

    Directory of Open Access Journals (Sweden)

    A. López San Román

    2005-04-01

    Full Text Available Aim: adherence to therapy is important to ensure success. We wanted to explore this feature in patients with inflammatory bowel disease. Patients and methods: we explored adherence to treatment and its modifiers in 40 patients with inflammatory bowel disease using a battery of tests. Results: a 67% of patients (95% CI: 51-81% acknowledged a certain degree of involuntary nonadherence, and 35% (95% CI: 20-51% of voluntary nonadherence. Overall, 72% (95% CI: 56-85% of patients had some form of nonadherence. An objective correlation of these self-reported data was assessed by the determination of urine salicylate levels in the subset of patients treated with mesalazine or its derivatives (15 cases. Two of them (13% had no detectable urinary drug levels, indicating complete nonadherence. Voluntary nonadherence was higher in patients with lower scores in the intestinal (p = 0.02 and social areas (p = 0.015 of IBDQ-32, as well as in those with less active Crohn's disease (p Objetivo: la adhesión al tratamiento es importante para el éxito del mismo. Quisimos conocer este dato en pacientes con enfermedad inflamatoria intestinal. Pacientes y métodos: hemos explorado la adhesión al tratamiento y sus condicionantes en 40 pacientes con enfermedad inflamatoria intestinal, aplicando una batería de pruebas. Resultados: un 67% (IC 95%: 51-81% de los pacientes presentaba algún grado de falta involuntaria de adhesión. A su vez, un 35% (20-51% de los pacientes presentaba algún grado de falta voluntaria de adhesión. Globalmente, un 72% (56-85% de los pacientes presentó uno u otro tipo de falta de adhesión. El correlato objetivo, se obtuvo mediante la determinación de salicilatos en orina en el subgrupo de pacientes bajo tratamiento con mesalazina o derivados (15 casos. Dos de ellos (13%, no tenían niveles detectables, traduciendo falta absoluta de adhesión al tratamiento. La adhesión intencionada era tanto más baja cuanto menores eran las

  8. Impact of free-living amoebae on presence of Parachlamydia acanthamoebae in the hospital environment and its survival in vitro without requirement for amoebae.

    Science.gov (United States)

    Fukumoto, Tatsuya; Matsuo, Junji; Hayashi, Yasuhiro; Hayashi, Masahiro; Oguri, Satoshi; Nakamura, Shinji; Mizutani, Yoshihiko; Yao, Takashi; Akizawa, Kouzi; Suzuki, Haruki; Shimizu, Chikara; Matsuno, Kazuhiko; Yamaguchi, Hiroyuki

    2010-09-01

    Parachlamydia acanthamoebae is an obligately intracellular bacterium that infects free-living amoebae and is a potential human pathogen in hospital-acquired pneumonia. We examined whether the presence of P. acanthamoebae is related to the presence of Acanthamoeba in an actual hospital environment and assessed the in vitro survival of P. acanthamoebae. Ninety smear samples were collected between November 2007 and March 2008 (trial 1, n = 52) and between October 2008 and February 2009 (trial 2, n = 38) from the floor (dry conditions, n = 56) and sink outlets (moist conditions, n = 34) of a hospital. The prevalences of P. acanthamoebae DNA in the first and second trials were 64.3% and 76%, respectively. The prevalences of Acanthamoeba DNA in the first and second trials were 48% and 63.1%, respectively. A statistical correlation between the prevalence of P. acanthamoebae and that of Acanthamoeba was found (trial 1, P = 0.011; trial 2, P = 0.022), and that correlation increased when samples from just the dry area (floor smear samples, P = 0.002) were analyzed but decreased when samples from a moist area were analyzed (P = 0.273). The in vitro experiment showed that, without Acanthamoeba, P. acanthamoebae could not survive in dry conditions for 3 days at 30 degrees C or 15 days at 15 degrees C. Thus, both organisms were coincidentally found in an actual hospital environment, with the presence of Acanthamoeba having a significant effect on the long-term survival of P. acanthamoebae, suggesting that this potential human pathogen could spread through a hospital environment via Acanthamoeba.

  9. A population-based study on the incidence of inflammatory bowel disease in Oviedo (Northern Spain Incidencia de la enfermedad inflamatoria intestinal (EII en población general en el área de Oviedo

    Directory of Open Access Journals (Sweden)

    L. Rodrigo

    2004-05-01

    Full Text Available Objective: to assess the incidence of inflammatory bowel disease in Oviedo (Northern Spain, and to describe the clinical features of new patients. Patients and methods: a prospective population-based study was made at the Health Area IV, Principality of Asturias (Oviedo, 312,324 inhabitants. All new diagnosed patients with inflammatory bowel disease were registered over a 2-year period. Results: a total of 85 patients were included, 47 of these with ulcerative colitis (UC, 37 with Crohn's disease (CD, and 1 with undetermined colitis. The overall adjusted incidence rate of UC and CD per 10(5 inhabitants between 15-64 years was 9.1 (95% CI: 5-13.1 and 7.5 (95% CI: 3.8-11.2, respectively. The global male/female ratio was 0.9, without significant differences between both diseases. CD patients were younger than those with UC (33 ± 15 years vs 45 ± 20 years; p Objetivo: conocer la incidencia de enfermedad inflamatoria intestinal en el área de Oviedo y describir las características clínicas de los nuevos pacientes. Pacientes y métodos: estudio prospectivo poblacional en el Área Sanitaria IV del Principado de Asturias (Oviedo, 312.324 habitantes. Fueron registrados todos los pacientes nuevos diagnosticados de enfermedad inflamatoria intestinal en un periodo de 2 años consecutivos. Resultados: se incluyeron un total de 85 pacientes, 47 con colitis ulcerosa (CU, 37 con enfermedad de Crohn (EC y 1 con colitis indeterminada (CI. La tasa de incidencia ajustada por 10(5 habitantes entre 15 y 64 años, de CU y EC, fue de 9,1 para CU (IC95%: 5-13,1 y 7,5 para la EC (IC95%: 3,8-11,2. La proporción hombre/mujer fue de 0,9, sin diferencias significativas entre ambas enfermedades. Los pacientes con EC eran más jóvenes que los que tenían CU (33 ± 15 años vs 45 ± 20 años; p<0,05. La mayor parte de los pacientes con EC tenían menos de 35 años (65%, mientras que los que presentaban CU fueron diagnosticados entre los 25 y 64 años (81%. La extensi

  10. MODULACION DE LA EXPRESION DE INTERLEUCINA-6 EN RESPUESTA AL MOVIMIENTO DENTARIO INDUCIDO POR EL ARCO .014 DE NiTi EN PACIENTES CON MALOCLUSION

    OpenAIRE

    Leon Romero, Luis Cenen

    2012-01-01

    El apiñamiento dentario es una anomalía dentomaxilar altamente frecuente, asociado con otros signos de maloclusión. El recurso clínico-ortodóntico para corregirlo es el movimiento dentario inducido por fuerzas mecánicas en los tejidos periodontales que bajo la fuerza mantenida, sufren una serie de modificaciones reguladas por procesos histológicos y fisiológicos que pueden dar como respuesta reacciones inflamatorias. La biología del movimiento dentario comprende el estudio de l...

  11. Función barrera epitelial en un modelo de disfunción intestinal inducido por parasitosis con Trichinella spiralis en la rata

    OpenAIRE

    Fernández Blanco, Joan Antoni

    2013-01-01

    La función barrera intestinal constituye la primera línea de defensa del tracto digestivo. Ésta se ve condicionada por el transporte hidroelectrolítico, la permeabilidad y la motilidad intestinales. Alteraciones de estas funciones favorecen la estimulación continua, por antígenos y microorganismos luminales, del sistema inmune local llevando a estados inflamatorios más o menos crónicos. Estos cambios parecen caracterizar diversas patologías inflamatorias y funcionales gastrointestinales, incl...

  12. MODULACION DE LA EXPRESION DE INTERLEUCINA-6 EN RESPUESTA AL MOVIMIENTO DENTARIO INDUCIDO POR EL ARCO .014 DE NiTi EN PACIENTES CON MALOCLUSION

    OpenAIRE

    Leon Romero, Luis Cenen

    2012-01-01

    El apiñamiento dentario es una anomalía dentomaxilar altamente frecuente, asociado con otros signos de maloclusión. El recurso clínico-ortodóntico para corregirlo es el movimiento dentario inducido por fuerzas mecánicas en los tejidos periodontales que bajo la fuerza mantenida, sufren una serie de modificaciones reguladas por procesos histológicos y fisiológicos que pueden dar como respuesta reacciones inflamatorias. La biología del movimiento dentario comprende el estudio de l...

  13. Interferon beta-1a in chronic inflammatory demyelinating polyneuropathy: case report Interferon beta en polineuropatía crónica inflamatoria desmienlinizante: caso clínico

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    Andrés Maria Villa

    2004-09-01

    Full Text Available Chronic inflammatory demyelinating polyneuropathy (CIDP is an acquired immune-mediated neuropathy. It presents with a course of progression which may be slow and steady or step-wise or relapsing. Sensory ataxic polyneuropathy may be the only clinical manifestation of this disease. Treatment with interferon beta1a (INF beta1a has been tried with different results in patients who were refractory to other, more conventional, immunomodulatory therapies. Here we report on a patient who had a relapsing form of pure sensory ataxic CIDP and who failed to respond to intravenous human immunoglobulin. He was put on INF beta1a for 3 years. During this period he suffered no relapses while his condition stabilized.La polineuropatía crónica inflamatoria desmielinizante (PCID es una neuropatía inmuno-mediada, que presenta un curso clínico primariamente progresivo o en forma de recaídas. Las manifestaciones sensoriales pueden ser su unica forma de expresión clínica. El tratamiento con interferon beta 1a (IFN beta1a ha sido ensayado en varias oportunidades, con diferentes respuestas terapéuticas, en pacientes refractarios a las terapias inmunomoduladoras convencionales. Nosotros comunicamos un paciente con una forma ataxica recurrente de PCID, que no respondió al tratamiento con inmunoglobulina endovenosa. Posteriormente fue tratado con IFN beta 1 a por tres años. Durante el período de seguimiento no mostró nuevas recaídas y su cuadro neurológico se estabilizó.

  14. Papel de la ventilación mecánica durante la circulación extracorporea sobre los mediadores de la respuesta inflamatoria

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    López Gomez, Amparo

    2015-01-01

    RESUMEN TESIS: PAPEL DE LA VENTILACIÓN MECÁNICA DURANTE LA CIRCULACIÓN EXTRACORPÓREA SOBRE LOS MEDIADORES DE LA RESPUESTA INFLAMATORIA. La Cardiopatía Isquémica (CI) representa la mayor causa de mortalidad en países desarrollados. En España, supone el mayor número de muertes cardiovasculares, siendo el infarto agudo de miocardio (IMA) la enfermedad isquémica más frecuente. Datos recientes sugieren que se ha producido un aumento de la prevalencia de pacientes con CI . La revasculariza...

  15. Cooperación oncogénica entre la citoquina pro-inflamatoria SP y el receptor tirosina quinasa ErbB-2/HER2

    OpenAIRE

    Garcia Recio, Susana

    2012-01-01

    INTRODUCCIÓN: La inflamación es un sello distintivo del cáncer. La inflamación crónica está fuertemente relacionada con el desarrollo y la progresión tumoral y suministra al tumor múltiples citoquinas proinflamatorias que pueden modular la señalización de la célula tumoral. Este es el caso de la Sustancia P (SP), una citoquina pro-inflamatoria/neuropéptido presente en el microambiente tumoral perteneciente a la familia de las taquiquininas y que señaliza principalmente a través de su receptor...

  16. Situaciones especiales en la enfermedad inflamatoria intestinal: primer consenso latinoamericano de la Pan American Crohn's and Colitis Organisation (PANCCO (Segunda parte

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    J.K. Yamamoto-Furusho

    2017-04-01

    Full Text Available Este es el primer Consenso Latinoamericano de la Pan American Crohn's and Colitis Organisation (PANCCO que corresponde a situaciones especiales en pacientes con enfermedad inflamatoria intestinal (EII. El objetivo de este consenso es concientizar a la comunidad médica de todos los países de América Latina acerca del embarazo, la vacunación, las infecciones y las neoplasias, incluyendo el cáncer colorrectal, así como los aspectos pediátricos en pacientes con EII.

  17. Consecuencias clínicas de la persistencia de actividad inflamatoria en la arteritis de células gigantes. Estudio de factores implicados

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    García Martínez, Ana

    2010-01-01

    La arteritis de células gigantes (ACG) afecta arterias de mediano y gran calibre. La inflamación de la carótida y sus ramas da lugar a los síntomas craneales típicos de la enfermedad. Las lesiones inflamatorias expresan gran cantidad de mediadores que provocan la muerte celular y la destrucción de fibras elásticas. El remodelado de la pared arterial puede dar lugar a alteraciones estructurales vasculares, como la aparición de estenosis arteriales que producen síntomas isquémicos o el desarrol...

  18. Moléculas de adhesión endotelial: nuevas dianas terapéuticas en la enfermedad inflamatoria intestinal

    OpenAIRE

    Soriano Izquierdo, Antonio

    2004-01-01

    INTRODUCCIÓN: El tratamiento de la enfermedad inflamatoria intestinal se ha basado en el uso de aminosalicilatos corticoides e inmunosupresores, que ayudan a controlar los brotes de la enfermedad, aunque no consiguen su curación. El bloqueo de las moléculas de adhesión endotelial, que permiten el reclutamiento de leucocitos desde el lecho vascular hacia el área inflamada, podría ser una diana terapéutica útil en esta enfermedad.OBJETIVOS: 1. Determinar el valor de las moléculas de adhesión en...

  19. Papel de la ventilación mecánica durante la circulación extracorporea sobre los mediadores de la respuesta inflamatoria

    OpenAIRE

    López Gomez, Amparo

    2015-01-01

    RESUMEN TESIS: PAPEL DE LA VENTILACIÓN MECÁNICA DURANTE LA CIRCULACIÓN EXTRACORPÓREA SOBRE LOS MEDIADORES DE LA RESPUESTA INFLAMATORIA. La Cardiopatía Isquémica (CI) representa la mayor causa de mortalidad en países desarrollados. En España, supone el mayor número de muertes cardiovasculares, siendo el infarto agudo de miocardio (IMA) la enfermedad isquémica más frecuente. Datos recientes sugieren que se ha producido un aumento de la prevalencia de pacientes con CI . La revasculariza...

  20. Acanthamoeba misidentification and multiple labels: redefining genotypes T16, T19, and T20 and proposal for Acanthamoeba micheli sp. nov. (genotype T19).

    Science.gov (United States)

    Corsaro, Daniele; Walochnik, Julia; Köhsler, Martina; Rott, Marilise B

    2015-07-01

    Acanthamoeba species are ubiquitous amoebae able to cause important infections in humans and other vertebrates. The full/near-full sequences (>2000 bp) of the small subunit ribosomal RNA gene (SSU rDNA or 18S rDNA) are used to cluster Acanthamoeba as genotypes, labeled T1 to T20. Genotype T15 remains an exception, being described only partially on a 1500-bp fragment. Strains are thus usually identified based on their 18S identity matches with reference strains, often using shorter (Acanthamoeba micheli sp. nov., for this strain. Furthermore, partial molecular phylogenies were performed to show that several other misassigned T16 partial sequences belong to a new genotype. This latter includes also misassigned T4 partial sequences, only recently available as full sequences and labeled as T20. We thus reassign these partial sequences to the genotype T20. Longer sequences, ideally at least 90 % of the total gene length, should be obtained from strains to ensure reliable diagnostic and phylogenetic results.

  1. Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus

    DEFF Research Database (Denmark)

    Hansen, Birgitte Rønde; Kronborg, Gitte

    2003-01-01

    Acanthamoeba keratitis is potentially blinding and often associated with contact lens wearing. A human immunodeficiency virus (HIV)-positive patient, a non-contact lens wearer, presented with keratitis. She experienced a protracted course of disease, characterized by exacerbations and remissions...

  2. In vitro effect of Acanthospermum australe (Asteraceae extracts on Acanthamoeba polyphaga trophozoites

    Directory of Open Access Journals (Sweden)

    LC Castro

    2013-01-01

    Full Text Available Acanthamoeba is a free-living protozoan widely distributed in the environment, occurring in vegetative trophozoite and resistance cyst stages during its life cycle. It constitutes an etiological agent of Acanthamoeba keratitis, a disease that may cause severe ocular inflammation and blindness. New drugs can be developed from molecules found in plants and thus help in its difficult treatment. Acanthospermum australe (Asteraceae, a plant used in folk medicine, had its effect tested on Acanthamoeba polyphaga. Aqueous and ethanolic extracts of A. austral were obtained from aerial parts for infusion and static maceration, respectively. Concentrations of 10, 5, 2.5, 1.25 and 0.625 mg/ml of the extract were tested against Acanthamoeba polyphaga trophozoites. The cytotoxic effect of the extracts was tested in mammalian cells using the 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT assay. RESULTS: The 10 mg/ml concentration of ethanolic extract was lethal to 100% of the A. polyphaga trophozoites in 24 h and both extracts presented cytotoxic effect against mammalian cells. These findings suggest that the A. austral ethanolic extract may have compounds with relevance to the development of new amoebicidal drugs.

  3. Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii.

    Science.gov (United States)

    Lee, Yu-Ran; Na, Byoung-Kuk; Moon, Eun-Kyung; Song, Su-Min; Joo, So-Young; Kong, Hyun-Hee; Goo, Youn-Kyoung; Chung, Dong-Il; Hong, Yeonchul

    2015-01-01

    Encystation of Acanthamoeba leads to the formation of resilient cysts from vegetative trophozoites. This process is essential for parasite survival under unfavorable conditions such as starvation, low temperatures, and exposure to biocides. During encystation, a massive turnover of intracellular components occurs, and a large number of organelles and proteins are degraded by proteases. Previous studies with specific protease inhibitors have shown that cysteine and serine proteases are involved in encystation of Acanthamoeba, but little is known about the role of metalloproteases in this process. Here, we have biochemically characterized an M17 leucine aminopeptidase of Acanthamoeba castellanii (AcLAP) and analyzed its functional involvement in encystation of the parasite. Recombinant AcLAP shared biochemical properties such as optimal pH, requirement of divalent metal ions for activity, substrate specificity for Leu, and inhibition profile by aminopeptidase inhibitors and metal chelators with other characterized M17 family LAPs. AcLAP was highly expressed at a late stage of encystation and mainly localized in the cytoplasm of A. castellanii. Knockdown of AcLAP using small interfering RNA induced a decrease of LAP activity during encystation, a reduction of mature cyst formation, and the formation of abnormal cyst walls. In summary, these results indicate that AcLAP is a typical M17 family enzyme that plays an essential role during encystation of Acanthamoeba.

  4. Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii.

    Directory of Open Access Journals (Sweden)

    Yu-Ran Lee

    Full Text Available Encystation of Acanthamoeba leads to the formation of resilient cysts from vegetative trophozoites. This process is essential for parasite survival under unfavorable conditions such as starvation, low temperatures, and exposure to biocides. During encystation, a massive turnover of intracellular components occurs, and a large number of organelles and proteins are degraded by proteases. Previous studies with specific protease inhibitors have shown that cysteine and serine proteases are involved in encystation of Acanthamoeba, but little is known about the role of metalloproteases in this process. Here, we have biochemically characterized an M17 leucine aminopeptidase of Acanthamoeba castellanii (AcLAP and analyzed its functional involvement in encystation of the parasite. Recombinant AcLAP shared biochemical properties such as optimal pH, requirement of divalent metal ions for activity, substrate specificity for Leu, and inhibition profile by aminopeptidase inhibitors and metal chelators with other characterized M17 family LAPs. AcLAP was highly expressed at a late stage of encystation and mainly localized in the cytoplasm of A. castellanii. Knockdown of AcLAP using small interfering RNA induced a decrease of LAP activity during encystation, a reduction of mature cyst formation, and the formation of abnormal cyst walls. In summary, these results indicate that AcLAP is a typical M17 family enzyme that plays an essential role during encystation of Acanthamoeba.

  5. Cytotoxic effect of organic solvents and surfactant agents on Acanthamoeba castellanii cysts.

    Science.gov (United States)

    Ezz Eldin, Hayam Mohamed; Sarhan, Rania Mohamed

    2014-05-01

    Acanthamoeba castellanii is a protozoan parasite that may cause sight-threatening keratitis in some individuals. Its eradication is difficult because the trophozoites encyst making organisms highly resistant to anti-amoebic drugs. To test new anti-Acanthamoeba agents, usually having low water solubility, organic solvents and surfactant agents should be used. Therefore, the lethal effect of different concentrations of the solvents acetone, methanol, ethanol, and DMSO and surfactant agents Tween 20, Tween 80, and Triton X-100 was tested. The minimal inhibitory concentrations (MIC) were determined against Acanthamoeba cysts. Results of the present study showed that the MIC for ethanol, methanol, acetone and DMSO was 25, 12.5, 12.5, and 10%, respectively and for Tween 20, Tween 80, and Triton X-100 was 0.25, 0.06, and 0.03%, respectively. There was no significant inhibitory effect on the multiplication of Acanthamoeba cysts as compared to parasite control when using the concentrations 3.12% for ethanol, 1.6% for methanol and acetone, 1.25% for DMSO, and 0.016% for Tween 20. On the other hand, both Tween 80 and Triton X-100 showed highly significant difference in comparison to parasite control almost among all the range of concentrations used in this study, and both showed lethal effect of 19 and 27.2%, respectively at their least concentration.

  6. Molecular Identification and Sequencing of Mannose Binding Protein (MBP Gene of Acanthamoeba palestinensis

    Directory of Open Access Journals (Sweden)

    M Rezaeian

    2010-02-01

    Full Text Available "nBackground: Acanthamoeba keratitis develops by pathogenic Acanthamoeba such as A. pal­es­tinen­sis. Indeed this species is one of the known causative agents of amoebic keratitis in Iran. Mannose Binding Protein (MBP is the main pathogenicity factors for developing this sight threatening disease. We aimed to characterize MBP gene in pathogenic Acanthamoeba isolates such as A. palestinensis."nMethods: This experimental research was performed in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2007-2008.  A. palestinensis was grown on 2% non-nutrient agar overlaid with Escherichia coli. DNA extraction was performed using phenol-chloroform method. PCR reaction and amplification were done using specific primer pairs of MBP. The amplified fragment were purified and sequenced. Finally, the obtained fragment was deposited in the gene data bank."nResults: A 900 bp PCR-product was recovered after PCR reaction. Sequence analysis of the purified PCR product revealed a gene with 943 nucleotides. Homology analysis of the ob­tained sequence showed 81% similarity with the available MBP gene in the gene data bank. The fragment was deposited in the gene data bank under accession number EU678895"nConclusion: MBP is known as the most important factor in Acanthamoeba pathogenesis cas­cade. Therefore, characterization of this gene can aid in developing better therapeutic agents and even immunization of high-risk people.

  7. Impression Cytology in Eyes with Clinical and Confocal Scan Features of Acanthamoeba Keratitis

    Directory of Open Access Journals (Sweden)

    Mozhgan Rezaei Kanavi

    2013-01-01

    Full Text Available Purpose: To report impression cytology findings in specimens obtained from eyes with clinical and confocal microscopic features of Acanthamoeba keratitis (AK. Methods: In this interventional case series, impression cytology was obtained from corneas of patients with clinical and confocal microscopic features indicative of AK. Specimens were stained with Periodic acid-Schiff/Papanicolaou (PAS/PAP and examined for the presence of PAS-reactive Acanthamoeba cysts and/or hyperchromatic pear-shaped trophozoites. All specimens were then decolorized and re-stained with calcofluor white (CFW for the presence of chemofluorescent cysts. Results: Fifty-six eyes of 50 patients with mean age of 25.5±9.3 (range, 17 to 78 years were evaluated. Forty-one (82% cases were female and 51 (91.1% eyes had history of contact lens wear. PAS-reactive Acanthamoeba cysts and/or hyperchromatic pear-shaped trophozoites were identified in 53 eyes (94.6%, 2 of which demonstrated only trophozoitelike structures. CFW staining was able to reveal the presence of chemofluorescent cysts in all 51 specimens (91.1% in which cysts had been demonstrated with PAS/PAP staining. Trophozoites were not detected with CFW due to background staining of the cellulose acetate strip used for impression cytology. Conclusion: Corneal impression cytology, stained with PAS/PAP or with CFW, successfully detects Acanthamoeba and can be employed for early noninvasive diagnosis of AK.

  8. Occurrence of pathogenic Acanthamoeba genotypes in nasal swabs of cancer patients in Iran.

    Science.gov (United States)

    Memari, Fatemeh; Niyyati, Maryam; Haghighi, Ali; Seyyed Tabaei, Seyyed Javad; Lasjerdi, Z

    2015-05-01

    Incidences of Acanthamoeba granulomatous encephalitis (AGE) have been increased due to a rise in the number of high-risk people, such as immunodeficient patients. Indeed, immunosuppress situation can render the patient in acquiring opportunistic Acanthamoeba infections. In this study, analysis was carried out to verify the presence of free-living amoebae of Acanthamoeba genus in nasal swabs of cancer patients in hospitals of Tehran, Iran. Detection of isolates was based on morphotyping and PCR sequencing of the Diagnostic Fragment 3 (DF3) to identify strains at the genotype level. In addition, the pathogenic potential of the isolates was assayed using temperature and osmotolerance assays. The obtained results revealed that nine isolated strains belonging to T4 genotype-exhibited pathogenic potential. After sequencing, genotype T4 was found to be the most common one in the samples included in this study. Genotype T3 and T5 were also identified. To the best of our knowledge, this is the first study on the typing of Acanthamoeba strains at the genotype level in cancer patients in Iran and worldwide.

  9. Microarray and KOG analysis of Acanthamoeba healyi genes up-regulated by mouse-brain passage.

    Science.gov (United States)

    Moon, Eun-Kyung; Xuan, Ying-Hua; Kong, Hyun-Hee

    2014-08-01

    Long-term cultivation in a laboratory could reduce the virulence of Acanthamoeba. To identify virulence factors of Acanthamoeba, the authors compared the transcription profiles of long-term cultivated Acanthamoeba healyi (OLD) and three times mouse-brain passaged A. healyi (MBP) using microarray analysis and eukaryotic orthologous group (KOG) assignments. Microarray analysis revealed that 601 genes were up-regulated by mouse-brain passage. The results of real-time PCR of 8 randomly selected genes up-regulated in the MBP strain confirmed microarray analysis findings. KOG assignments showed relatively higher percentages of the MBP strain up-regulated genes in T article (signal transduction mechanism), O article (posttranslational modification, protein turnover, chaperones), C article (energy production and conversion), and J article (translation, ribosomal structure and biogenesis). In particular, the MBP strain showed higher expressions of cysteine protease and metalloprotease. A comparison of KOG assignments by microarray analysis and previous EST (expressed sequence tags) analysis showed similar populations of up-regulated genes. These results provide important information regarding the identification of virulence factors of pathogenic Acanthamoeba.

  10. Isolation and identification of Acanthamoeba spp. from thermal swimming pools and spas in Southern Brazil.

    Science.gov (United States)

    Fabres, Laura Fuhrich; Rosa Dos Santos, Sayonara Peixoto; Benitez, Lisianne Brittes; Rott, Marilise Brittes

    2016-03-01

    Free-living amoebae (FLA) are widely distributed in soil and water. A few number of them are implicated in human disease: Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris and Sappinia diploidea. Species of Acanthamoeba can cause keratitis and brain infections. In this study, 72 water samples were taken from both hot tubs and thermal swimming pools in the city of Porto Alegre, RS, Brazil, to determine the presence of Acanthamoeba in the water as well as perform the phenotypic and genotypic characterization of the isolates. The identification of the isolates was based on the cysts morphology and PCR amplification using genus-specific oligonucleotides. When the isolates were submitted to PCR reaction only 8 were confirmed as belonging to the genus Acanthamoeba. The sequences analysis when compared to the sequences in the GenBank, showed genotype distribution in group T3 (12,5%), T5 (12,5%), T4 (25%) and T15 (50%). The results of this study confirmed the presence of potentially pathogenic isolates of free living amoebae in hot swimming pool and spas which can present risks to human health.

  11. Efficacy of Korean Multipurpose Contact Lens Disinfecting Solutions against Acanthamoeba castellanii

    Science.gov (United States)

    Moon, Eun-Kyung; Park, Hye-Ryun; Quan, Fu-Shi; Kong, Hyun-Hee

    2016-01-01

    Acanthamoeba keratitis has been increasing in recent years. Main risk factors are contact lens wear and their cleaning solutions. Most contact lens wearers use multipurpose disinfecting solutions (MPDS) for cleansing and disinfecting microorganisms because of its convenience. We determined amoebicidal effects of MPDS made in Korea and their cytotoxicity on human corneal epithelium cells. Fifteen commercial MPDS (A to O) were tested for their amoebicidal effects on Acanthamoeba castellanii trophozoites and cysts by using a most probable number (MPN) technique. Among them, 7 kinds of MPDS showed little or no amoebicidal effects for 24 hr exposure. Solutions A, B, G, H, L, and O showed positive amoebicidal effects, and solutions M and N killed almost all trophozoites and cysts after 24 hr exposure. However, 50%-N solution showed 56% cytotoxicity on human corneal epithelial cells within 4 hr exposure, and 50%-O solution also showed 62% cytotoxicity on human cells within 4 hr exposure. Solution A did not show any cytotoxicity on human cells. These results revealed that most MPDS made in Korea were ineffective to kill Acanthamoeba. The solutions having amoebicidal activity also showed high levels of cytotoxicity on human corneal epithelial cells. New formulations for improved MPDS that are amoebicidal but safe for host cells are needed to prevent Acanthamoeba keratitis. PMID:28095653

  12. Genotyping of potentially pathogenic Acanthamoeba strains isolated from nasal swabs of healthy individuals in Peru.

    Science.gov (United States)

    Cabello-Vílchez, Alfonso Martín; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Reyes-Batlle, María; González, Ana C; Guerra, Humberto; Gotuzzo, Eduardo; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-02-01

    Free Living Amoebae (FLA) of Acanthamoeba genus are widely distributed in the environment and can be found in the air, soil and water; and have also been isolated from air-conditioning units. In humans, they are causative agents of a sight-threating infection of the cornea, Acanthamoeba keratitis (AK) and a fatal infection of the central nervous system known as Granulomatous Amoebic Encephalitis (GAE). In this study, a survey was conducted in order to determine the presence and pathogenic potential of free-living amoebae of Acanthamoeba genus in nasal swabs from individuals in two regions of Peru. Identification of isolates was based on cyst morphology and PCR-sequencing of the Diagnostic Fragment 3 to identify strains at the genotype level. The pathogenic potential of the isolates was also assayed using temperature and osmotolerance assays and extracellular proteases zymograms. The obtained results revealed that all isolated strains exhibited pathogenic potential. After sequencing the highly variable DF3 (Diagnostic Fragment 3) region in the 18S rRNA gene as previously described, genotype T4 was found to be the most common one in the samples included in this study but also genotype T15 was identified. To the best of our knowledge, this is the first study on the characterization of Acanthamoeba strains at the genotype level and the first report of genotype T4 and T15 in Peru.

  13. Human IgA inhibits adherence of Acanthamoeba polyphaga to epithelial cells and contact lenses.

    Science.gov (United States)

    Campos-Rodríguez, Rafael; Oliver-Aguillón, Gabriela; Vega-Pérez, Luz M; Jarillo-Luna, Adriana; Hernández-Martínez, Dolores; Rojas-Hernández, Saúl; Rodríguez-Monroy, Marco A; Rivera-Aguilar, Víctor; González-Robles, Arturo

    2004-09-01

    Specific anti-Acanthamoeba IgA antibodies have been detected in the serum and tears of patients and healthy individuals. However, the role of human secretory IgA antibodies in inhibiting the adherence of Acanthamoeba had not been previously investigated. Therefore, the purpose of this study was to purify secretory IgA from human colostrum and analyze its effect on the adherence of Acanthamoeba trophozoites to contact lenses and Madin-Darby canine kidney (MDCK) cells. IgA antibodies to Acanthamoeba polyphaga in colostrum of healthy women as well as in saliva and serum of healthy subjects were analyzed by ELISA and Western blot analysis. In serum, saliva, and colostrum, we detected IgA antibodies that recognized several antigens of A. polyphaga. In addition, colostrum and IgA antibodies purified from it inhibited adherence of A. polyphaga trophozoites to contact lenses and MDCK cells. These results suggest that IgA antibodies may participate in the resistance to the amoebic infection, probably by inhibiting the adherence of the trophozoites to contact lenses and corneal epithelial cells.

  14. Acanthamoeba polyphaga is a possible host for Vibrio cholerae in aquatic environments.

    Science.gov (United States)

    Sandström, Gunnar; Saeed, Amir; Abd, Hadi

    2010-09-01

    Acanthamoeba is a genus of free-living amoebae found to be able to host many bacterial species living in the environment. Acanthamoebae and Vibrio cholerae are found in the aquatic environments of cholera endemic areas. Previously it has been shown that V. cholerae O1 and O139 can survive and grow in Acanthamoeba castellanii. The aim of this study was to examine the ability of Acanthamoeba polyphaga to host V. cholerae O1 and O139. The interaction between A. polyphaga and V. cholerae strains was studied by means of viable amoeba cell counts and viable count of the bacteria in the absence and presence of amoebae. The viable count of intracellularly growing bacteria was estimated by utilizing gentamicin assay. Electron microscopy was used to determine the localization of V. cholerae inside A. polyphaga. The results showed that A. polyphaga enhanced growth and survival of V. cholerae, which grew and survived inside the amoeba cells for 2weeks. The electron microscopy showed that A. polyphaga hosted intracellular V. cholerae localized in the vacuoles of amoeba cell. Neither the presence of V. cholerae together with A. polyphaga nor the intracellular localization of the bacteria inhibited growth and survival of A. polyphaga. The outcome of the interaction between these microorganisms may support strongly the role of A. polyphaga as host for V. cholerae O1 and O139.

  15. Free-living Amoebae (FLA: morphological and molecular identification of Acanthamoeba in dental unit water

    Directory of Open Access Journals (Sweden)

    Trabelsi H.

    2010-03-01

    Full Text Available The aim of our study was to detect free-living Amoebae (FLA by morphological methods and to identify Acanthamoeba spp. by PCR in the dental unit water lines (DUWL. Materials and methods: it was a prospective study dealing with 196 water samples collected from DUWL (94 samples taken in the early morning before materials flush and patient consultations and 102 samples taken after consultations. At the same time, 39 samples from tap water were realized. Results: 135 (69 % samples were positives by the morphological study with morphotypical diversity. The predominant morphotype was the monopodial (39.2 %. 18 strains of Acanthamoeba spp. were detected in DUW (13.3 % and three strains in tap water (10 %. The amplification of 18S rDNA gene of these strains of Acanthamoeba spp. was positive for all samples. Conclusion: the FLA and Acanthamoeba were isolated both in tap water and in dental unit. The amoeba pathogenicity has not been demonstrated after oral or dental contamination; but the presence of intracellular and pathogenic bacteria in the amoeba could be a source of microbiological risks for patients in case of deep dental care or immunodepression. The improvement of this dental unit was necessary by putting a filter of 0.2 microns porosity before the arrival of the water in hand-pieces allowing the limitation of FLA passage.

  16. Interactions between Human Norovirus Surrogates and Acanthamoeba spp.

    Science.gov (United States)

    Hsueh, Tun-Yun; Gibson, Kristen E

    2015-06-15

    Human noroviruses (HuNoVs) are the most common cause of food-borne disease outbreaks, as well as virus-related waterborne disease outbreaks in the United States. Here, we hypothesize that common free-living amoebae (FLA)-ubiquitous in the environment, known to interact with pathogens, and frequently isolated from water and fresh produce-could potentially act as reservoirs of HuNoV and facilitate the environmental transmission of HuNoVs. To investigate FLA as reservoirs for HuNoV, the interactions between two Acanthamoeba species, A. castellanii and A. polyphaga, as well as two HuNoV surrogates, murine norovirus type 1 (MNV-1) and feline calicivirus (FCV), were evaluated. The results showed that after 1 h of amoeba-virus incubation at 25°C, 490 and 337 PFU of MNV-1/ml were recovered from A. castellanii and A. polyphaga, respectively, while only few or no FCVs were detected. In addition, prolonged interaction of MNV-1 with amoebae was investigated for a period of 8 days, and MNV-1 was demonstrated to remain stable at around 200 PFU/ml from day 2 to day 8 after virus inoculation in A. castellanii. Moreover, after a complete amoeba life cycle (i.e., encystment and excystment), infectious viruses could still be detected. To determine the location of virus associated with amoebae, immunofluorescence experiments were performed and showed MNV-1 transitioning from the amoeba surface to inside the amoeba over a 24-h period. These results are significant to the understanding of how HuNoVs may interact with other microorganisms in the environment in order to aid in its persistence and survival, as well as potential transmission in water and to vulnerable food products such as fresh produce.

  17. Análisis de los ingresos hospitalarios por asma: características y factores pronóstico

    OpenAIRE

    Calvo Álvarez, Uxío

    2016-01-01

    El asma es una enfermedad inflamatoria crónica de las vías respiratorias que se caracteriza por la aparición de episodios recurrentes y variables en su intensidad de síntomas respiratorios (tos, sibilancias, opresión torácica, disnea…), limitación al flujo aéreo e hiperreactividad bronquial, teniendo como sustrato patológico la presencia de inflamación a nivel bronquial. Se considera el asma un problema de salud pública que afecta a 300 millones de personas en el mundo, si b...

  18. Anti-amoebic properties of a Malaysian marine sponge Aaptos sp. on Acanthamoeba castellanii.

    Science.gov (United States)

    Nakisah, M A; Ida Muryany, M Y; Fatimah, H; Nor Fadilah, R; Zalilawati, M R; Khamsah, S; Habsah, M

    2012-03-01

    Crude methanol extracts of a marine sponge, Aaptos aaptos, collected from three different localities namely Kapas, Perhentian and Redang Islands, Terengganu, Malaysia, were tested in vitro on a pathogenic Acanthamoeba castellanii (IMR isolate) to examine their anti-amoebic potential. The examination of anti-Acanthamoebic activity of the extracts was conducted in 24 well plates for 72 h at 30 °C. All extracts possessed anti-amoebic activity with their IC(50) values ranging from 0.615 to 0.876 mg/mL. The effect of the methanol extracts on the surface morphology of A. castellanii was analysed under scanning electron microscopy. The ability of the extracts to disrupt the amoeba cell membrane was indicated by extensive cell's blebbing, changes in the surface morphology, reduced in cell size and with cystic appearance of extract-treated Acanthamoeba. Number of acanthapodia and food cup was also reduced in this Acanthamoeba. Morphological criteria of apoptosis in Acanthamoeba following treatment with the sponge's extracts was determined by acridine orange-propidium iodide staining and observed by fluorescence microscopy. By this technique, apoptotic and necrotic cells can be visualized and quantified. The genotoxic potential of the methanol extracts was performed by the alkaline comet assay. All methanol extracts used were significantly induced DNA damage compared to untreated Acanthamoeba by having high percentage of scores 1, 2, and 3 of the DNA damage. Results from cytotoxicity and genotoxicity studies carried out in the present study suggest that all methanol extracts of A. aaptos have anti-amoebic properties against A. castellanii.

  19. Evaluation of Ozone Application in Dental Unit Water Lines Con¬taminated with Pathogenic Acanthamoeba

    Directory of Open Access Journals (Sweden)

    Wafaa HIKAL

    2015-10-01

    Full Text Available Background: In this study morphological and molecular characterization of Acan­thamoeba strains, isolated from dental unit waterlines (DUWLs were surveyed and the levels of disinfection achievable in vitro by the application of ozone disinfect­ant to DUWLs were evaluate.Methods: Water samples were collected from air-water syringes, cup fillers and tap water before and at the end of the working day. They were cultured on non-nutrient agar (NNA plates. Species identification was carried out with a PCR assay based on sequence analysis of the 18S rRNA gene. The cellular response to ozone was tested on Acanthamoeba cyst with different doses at different contact time in vitro twice.Results: Prevalence rates for Acanthamoeba contamination were 100, 100 and 72% for air-water syringes, cup fillers and tap water, respectively. The morphological analysis revealed the presence of A. castellanii, A. griffin, A. hatchitti and A. lenticulata. Phylogenetic analysis of the sequences showed the four strains to be closely related to a sequence type (T3, T4, T5 and T11. Acanthamoeba cells were stained with try­pan blue, which revealed killed of Acanthamoeba instantaneously after 10 minutes in ozonized water. There was no growth of Acanthamoeba occurred after ozone treat­ment in water bottles for 5 minutes with a flow rate of 500 mg/hour.Conclusion: Ozone can play an important role in controlling the problem of contami­nation of DUWLs as a potent disinfectant.

  20. Genetic characterization of clinical acanthamoeba isolates from Japan using nuclear and mitochondrial small subunit ribosomal RNA.

    Science.gov (United States)

    Rahman, Md Moshiur; Yagita, Kenji; Kobayashi, Akira; Oikawa, Yosaburo; Hussein, Amjad I A; Matsumura, Takahiro; Tokoro, Masaharu

    2013-08-01

    Because of an increased number of Acanthamoeba keratitis (AK) along with associated disease burdens, medical professionals have become more aware of this pathogen in recent years. In this study, by analyzing both the nuclear 18S small subunit ribosomal RNA (18S rRNA) and mitochondrial 16S rRNA gene loci, 27 clinical Acanthamoeba strains that caused AK in Japan were classified into 3 genotypes, T3 (3 strains), T4 (23 strains), and T5 (one strain). Most haplotypes were identical to the reference haplotypes reported from all over the world, and thus no specificity of the haplotype distribution in Japan was found. The T4 sub-genotype analysis using the 16S rRNA gene locus also revealed a clear sub-conformation within the T4 cluster, and lead to the recognition of a new sub-genotype T4i, in addition to the previously reported sub-genotypes T4a-T4h. Furthermore, 9 out of 23 strains in the T4 genotype were identified to a specific haplotype (AF479533), which seems to be a causal haplotype of AK. While heterozygous nuclear haplotypes were observed from 2 strains, the mitochondrial haplotypes were homozygous as T4 genotype in the both strains, and suggested a possibility of nuclear hybridization (mating reproduction) between different strains in Acanthamoeba. The nuclear 18S rRNA gene and mitochondrial 16S rRNA gene loci of Acanthamoeba spp. possess different unique characteristics usable for the genotyping analyses, and those specific features could contribute to the establishment of molecular taxonomy for the species complex of Acanthamoeba.

  1. Status of free-living amoebae (Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris) in drinking water supplies in Karachi, Pakistan.

    Science.gov (United States)

    Yousuf, Farzana Abubakar; Siddiqui, Ruqaiyyah; Subhani, Faysal; Khan, Naveed Ahmed

    2013-06-01

    The ability of pathogenic free-living amoebae to produce infections is a growing concern. In this study, we investigated the presence of free-living amoebae (Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris) in drinking water supplies in Karachi, Pakistan. Fifty-two domestic tap water samples were examined. Amoebae were identified by morphological characteristics and polymerase chain reaction. Thirty percent of the examined samples were positive for Acanthamoeba spp., 8% for N. fowleri while B. mandrillaris were not recovered. Additionally we examined secretory IgA antibody to Acanthamoeba and B. mandrillaris. Acanthamoeba antibody prevalence rate was 100% in both males and females, while B. mandrillaris antibody prevalence rate was 5.5% in males only (females were negative). Our findings suggest that free-living amoebae are a potential health hazard in domestic water supplies in Karachi, Pakistan.

  2. Infection in a rat model reactivates attenuated virulence after long-term axenic culture of Acanthamoeba spp

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    Carolina De Marco Verissimo

    2013-11-01

    Full Text Available Prolonged culturing of many microorganisms leads to the loss of virulence and a reduction of their infective capacity. However, little is known about the changes in the pathogenic strains of Acanthamoeba after long culture periods. Our study evaluated the effect of prolonged culturing on the invasiveness of different isolates of Acanthamoeba in an in vivo rat model. ATCC strains of Acanthamoeba, isolates from the environment and clinical cases were evaluated. The in vivo model was effective in establishing the infection and differentiating the pathogenicity of the isolates and re-isolates. The amoebae cultured in the laboratory for long periods were less virulent than those that were recently isolated, confirming the importance of passing Acanthamoeba strains in animal models.

  3. Acanthamoeba genotypes T2, T4, and T11 in soil sources from El Hierro island, Canary Islands, Spain.

    Science.gov (United States)

    Reyes-Batlle, María; Zamora-Herrera, Jonadab; Vargas-Mesa, Alejandro; Valerón-Tejera, Marco Antonio; Wagner, Carolina; Martín-Navarro, Carmen Ma; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2016-08-01

    The genus Acanthamoeba includes pathogenic strains which are causative agents of keratitis and encephalitis that often may end fatal in humans and other animals. In the present study, forty soil samples were collected in the island of El Hierro, Canary Islands, Spain, and checked for the presence of Acanthamoeba. Samples were cultivated onto 2 % non-nutrient agar plates seeded with a layer of heat killed Escherichia coli. Amplification by PCR and sequencing of the DF3 region of the 18S rDNA of Acanthamoeba was carried out in order to confirm morphological identification of the amoebae. Furthermore, Acanthamoeba spp. was isolated from 47.5 % of soil samples. Moreover, genotypes T2, T4, and T11 were identified in these samples. To the best of our knowledge, this is the first study to establish genotypes T2, T4, and T11 in soil sources from El Hierro island.

  4. Necrotizing Meningoencephalitis in a Captive Black and White Ruffed Lemur (Varecia variegata variegata) Caused by Acanthamoeba T4 Genotype.

    Science.gov (United States)

    Gaide, N; Pelandakis, M; Robveille, C; Albaric, O; Jouvion, G; Souchon, M; Risler, A; Abadie, J

    2015-11-01

    A mature male, black and white ruffed lemur (Varecia variegata variegata) died in a zoological garden after a 4-day history of lethargy and non-responsive convulsions. Necropsy and histopathological examinations revealed acute necrotizing and haemorrhagic meningoencephalitis with intralesional amoebas confirmed by immunohistochemistry. Acanthamoeba T4 genotype was identified as the causative agent of the brain lesion, based on amplification and sequencing of 18S ribosomal RNA genes. The presence of free-living amoebas in water and mud from the lemur's environment was investigated by morphological and molecular analyses. The two predominant genera, representing 80% of isolated amoebas, were Naegleria spp. and Acanthamoeba spp. All Acanthamoeba isolates belonged to the T4 genotype. To the author's knowledge, this is the first report of a meningoencephalitis due to Acanthamoeba T4 genotype in Lemuridae with concurrent analysis of pathological tissues and environment.

  5. Infection in a rat model reactivates attenuated virulence after long-term axenic culture of Acanthamoeba spp

    Science.gov (United States)

    Veríssimo, Carolina De Marco; Maschio, Vinícius José; Correa, Ana Paula Folmer; Brandelli, Adriano; Rott, Marilise Brittes

    2013-01-01

    Prolonged culturing of many microorganisms leads to the loss of virulence and a reduction of their infective capacity. However, little is known about the changes in the pathogenic strains of Acanthamoeba after long culture periods. Our study evaluated the effect of prolonged culturing on the invasiveness of different isolates of Acanthamoeba in an in vivo rat model. ATCC strains of Acanthamoeba, isolates from the environment and clinical cases were evaluated. The in vivo model was effective in establishing the infection and differentiating the pathogenicity of the isolates and re-isolates. The amoebae cultured in the laboratory for long periods were less virulent than those that were recently isolated, confirming the importance of passing Acanthamoeba strains in animal models. PMID:24271042

  6. Análisis de la reacción inflamatoria y los procesos degenerativos provocados por la inyección intracerebroventricular de neuraminidasa en rata

    OpenAIRE

    Granados-Durán, Pablo

    2014-01-01

    Las infecciones víricas y bacterianas del sistema nervioso central (SNC) y la neuroinflamación han sido relacionadas en los últimos años con en el desarrollo de ciertas enfermedades neurodegenerativas, tales como la esclerosis múltiple, la enfermedad de Alzheimer, la esclerosis lateral amiotrófica o trastornos neurocognitivos como la demencia asociada al VIH o la esquizofrenia. Muchas de estas enfermedades continúan hoy sin tratamiento, incluyendo aquellas que son sospechosas de ser consecuen...

  7. Análisis de la reacción inflamatoria y los procesos degenerativos provocados por la inyección intracerebroventricular de neuraminidasa en rata

    OpenAIRE

    Granados-Durán, Pablo

    2015-01-01

    Las infecciones víricas y bacterianas del sistema nervioso central (SNC) y la neuroinflamación han sido relacionadas en los últimos años con en el desarrollo de ciertas enfermedades neurodegenerativas, tales como la esclerosis múltiple, la enfermedad de Alzheimer, la esclerosis lateral amiotrófica o trastornos neurocognitivos como la demencia asociada al VIH o la esquizofrenia. Muchas de estas enfermedades continúan hoy sin tratamiento, incluyendo aquellas que son sospechosas de ser consecuen...

  8. Hypoxia attenuates inflammatory mediators production induced by Acanthamoeba via Toll-like receptor 4 signaling in human corneal epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Hong [Department of Ophthalmology, Qilu Hospital, Shandong University, 107, Wenhua Xi Road, Jinan 250012 (China); The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, 107, Wenhua Xi Road, Jinan 250012 (China); Wu, Xinyi, E-mail: xywu8868@163.com [Department of Ophthalmology, Qilu Hospital, Shandong University, 107, Wenhua Xi Road, Jinan 250012 (China)

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer Hypoxia attenuates Acanthamoeba-induced the production of IL-8 and IFN-{beta}. Black-Right-Pointing-Pointer Hypoxia inhibits TLR4 expression in a time-dependent manner in HCECs. Black-Right-Pointing-Pointer Hypoxia inhibits Acanthamoeba-induced the activation of NF-{kappa}B and ERK1/2 in HCECs. Black-Right-Pointing-Pointer Hypoxia decreases Acanthamoeba-induced inflammatory response via TLR4 signaling. Black-Right-Pointing-Pointer LPS-induced the secretion of IL-6 and IL-8 is abated by hypoxia via TLR4 signaling. -- Abstract: Acanthamoeba keratitis (AK) is a vision-threatening corneal infection that is intimately associated with contact lens use which leads to hypoxic conditions on the corneal surface. However, the effect of hypoxia on the Acanthamoeba-induced host inflammatory response of corneal epithelial cells has not been studied. In the present study, we investigated the effect of hypoxia on the Acanthamoeba-induced production of inflammatory mediators interleukin-8 (IL-8) and interferon-{beta} (IFN-{beta}) in human corneal epithelial cells and then evaluated its effects on the Toll-like receptor 4 (TLR4) signaling, including TLR4 and myeloid differentiation primary response gene (88) (MyD88) expression as well as the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-{kappa}B) and extracellular signal-regulated kinases 1/2 (ERK1/2). We then studied the effect of hypoxia on a TLR4-specific inflammatory response triggered by the TLR4 ligand lipopolysaccharide (LPS). Our data showed that hypoxia significantly decreased the production of IL-8 and IFN-{beta}. Furthermore, hypoxia attenuated Acanthamoeba-triggered TLR4 expression as well as the activation of NF-{kappa}B and ERK1/2, indicating that hypoxia abated Acanthamoeba-induced inflammatory responses by affecting TLR4 signaling. Hypoxia also inhibited LPS-induced IL-6 and IL-8 secretion, myeloid differentiation primary response gene (88

  9. Reporting of T4 Genotype of Acanthamoeba Isolates in Recreational Water Sources of Gilan Province, Northern Iran

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    Maryam Niyyati

    2015-02-01

    Full Text Available Background: Acanthamoeba spp. is the causative agent of blindness keratitis and fatal encephalaitis. Presence of Acanthamoeba spp. in a wide variety of niches such as different water types can lead to exposure of high risk people such as contact lens wearers. The main aim of the present study was to explore the occurrence of Acanthamoeba genotypes in the recreational water sources using both morphological and molecular approaches in Gilan province, Iran.Materials and Methods: Overall, 50 samples were collected from recreational water sources including man- made and natural waters in Gilan province. Filtration and cultivation of samples was performed using non-nutrient agar. Cloning of Acanthamoeba spp. was done to eliminate bacterial and fungi contamination. PCR amplification and sequencing were performed using genus-specific primer pair. Genotype identification was based on homology analysis of 18S rRNA gene (DF3 of the obtained sequences with the available genes in the gene bank data base.Results: Out of 50 water samples, 15 (30% were positive for Acanthamoeba trophozoites and cysts according to morphological criteria. Cloning of 13 isolates (26% was done successfully. Molecular analysis of 13 Acanthamoeba strain revealed that all isolates were belonged to potentially pathogenic T4 genotype.Conclusion: T4 genotype is the main cause of Acanthamoeba-related infections. Presence of Acanthamoeba belonged to T4 genotype in recreational water sources is of concern for high risk people. Alarming sign and education to high risk people is of utmost importance to prevent such infections.

  10. Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples.

    Science.gov (United States)

    Kao, Po-Min; Tung, Min-Che; Hsu, Bing-Mu; Tsai, Hsien-Lung; She, Cheng-Yu; Shen, Shu-Min; Huang, Wen-Chien

    2013-03-01

    In this study, a quantitative real-time PCR was developed to detect and quantify Acanthamoeba spp. in various environmental water samples. The water samples were taken from watershed, water treatment plant, and three thermal spring recreation areas. The overall detection rate was 14.2 % (25/176) for Acanthamoeba spp. The percentages of samples containing Acanthamoeba spp. from river water, raw drinking water, and thermal spring water were 13 % (13/100), 25 % (7/28), and 10.4 % (5/48), respectively. Acanthamoeba spp. concentrations were determined according to SYBR Green quantitative real-time PCR. A plasmid-based standard curve was constructed to determine the Acanthamoeba concentration using dilution factors for achieving 1.36 × 10(9) gene copies per PCR for 18S rRNA gene in Acanthamoeba spp. The resulting concentrations varied by the type of water, in the range of 46-2.6 × 10(2) cells/l in positive raw drinking water, 2.7 × 10(2)-1.5 × 10(4) cells/l in river water, and 54-1.7 × 10(3) cells/l in thermal spring water. The presence of Acanthamoeba spp. in the raw drinking water samples was also found to have a significant difference with heterotrophic plate count. The presence of Acanthamoeba spp. in various aquatic environments may be a potential health hazard and must be further evaluated.

  11. Modelos animales para el estudio de la respuesta inflamatoria sistémica y de nutrición parenteral Animal models for the study of systemic inflammatory response and parenteral nutrition

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    J. M. Morán Penco

    2007-04-01

    Full Text Available El Síndrome de Respuesta Inflamatoria Sistémica (SRIS parece desencadenarse por la activación de un tipo de receptores llamados Toll-Like, propios de las células de respuesta inflamatoria y que, a través de señales citosólicas específicas, producen una cascada de reacciones que activan las Citocinas, Factores de Crecimiento y otros mediadores inflamatorios. En este trabajo, repasamos y discutimos varias clasificaciones de Modelos Animales para el estudio del SRIS y proponemos que estos modelos se dividan según los objetivos concretos a estudiar; que podrían ser los siguientes: 1º Para el estudio de los genes reguladores de los receptores Innatos y Adaptativos, de la respuesta inmunoinflamatoria. 2º Para el estudio de los receptores de señalización (Citocinas y Factores de Crecimiento. 3º Para el estudio de la respuesta frente a los señalizadores. 4º Para el estudio de tratamientos mediante bloqueo anti-inflamatorio específico (ILs, TNF y otros. 5º Modelos específicos de Sepsis. 6º Otros Modelos Inductores de SRIS. 7º Para el estudio de Modelos Terapéuticos diversos: -Tratamientos Anti-Inflamatorios. -Tratamientos Anticoagulantes: Inhibición de la Coagulación en ensayos Humanos-Fase II, con anticoagulantes: Antitrombina III, PCA y TFPI. -Tratamientos Antibióticos. -Tratamientos con Reposición de Volúmenes. -Tratamientos Quirúrgicos. En lo que respecta al apartado de Modelos animales para el estudio de la Nutrición Parenteral, podríamos hacer la siguiente clasificación y resumen de lo tratado: 1. Modelos Animales para el estudio de la Vía Parenteral de Administración. 2. Modelos para el estudio de la viabilidad, absorción y tolerancia locales en la vía de administración. 3. Modelos para el estudio de las Complicaciones. 4. Modelos animales para el estudio de la farmacodinamia, metabolización e investigación de la tolerancia de nuevas moléculas o substratos.The systemic inflammatory response syndrome (SRIS

  12. Detection of Vibrio cholerae and Acanthamoeba species from same natural water samples collected from different cholera endemic areas in Sudan

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    Saeed Amir

    2011-04-01

    Full Text Available Abstract Background Vibrio cholerae O1 and V. cholerae O139 infect humans, causing the diarrheal and waterborne disease cholera, which is a worldwide health problem. V. cholerae and the free-living amoebae Acanthamoeba species are present in aquatic environments, including drinking water and it has shown that Acanthamoebae support bacterial growth and survival. Recently it has shown that Acanthamoeba species enhanced growth and survival of V. cholerae O1 and O139. Water samples from different cholera endemic areas in Sudan were collected with the aim to detect both V. cholerae and Acanthamoeba species from same natural water samples by polymerase chain reaction (PCR. Findings For the first time both V. cholerae and Acanthamoeba species were detected in same natural water samples collected from different cholera endemic areas in Sudan. 89% of detected V. cholerae was found with Acanthamoeba in same water samples. Conclusions The current findings disclose Acanthamoedae as a biological factor enhancing survival of V. cholerae in nature.

  13. Immunological inter-strain crossreactivity correlated to 18S rDNA sequence types in Acanthamoeba spp.

    Science.gov (United States)

    Walochnik, J; Obwaller, A; Aspöck, H

    2001-02-01

    Various species of the genus Acanthamoeba have been described as potential pathogens; however, differentiation of acanthamoebae remains problematic. The genus has been divided into 12 18S rDNA sequence types, most keratitis causing strains exhibiting sequence type T4. We recently isolated a keratitis causing Acanthamoeba strain showing sequence type T6, but being morphologically identical to a T4 strain. The aim of our study was to find out, whether the 18S rDNA sequence based identification correlates to immunological differentiation. The protein and antigen profiles of the T6 isolate and three reference Acanthamoeba strains were investigated using two sera from Acanthamoeba keratitis patients and one serum from an asymptomatic individual. It was shown, that the T6 strain produces a distinctly different immunological pattern, while patterns within T4 were identical. Affinity purified antibodies were used to further explore immunological cross-reactivity between sequence types. Altogether, the results of our study support the Acanthamoeba 18S rDNA sequence type classification in the investigated strains.

  14. Isolation and genotyping of free-living environmental isolates of Acanthamoeba spp. from bromeliads in Southern Brazil.

    Science.gov (United States)

    Landell, Melissa Fontes; Salton, Juliana; Caumo, Karin; Broetto, Leonardo; Rott, Marilise B

    2013-07-01

    Species of Acanthamoeba are frequently isolated from distinct environmental sources such as water, soil, dust and air. They are responsible to cause infections and disease in humans and animals. In addition, Acanthamoeba sp. are considered an important reservoir of bacteria, virus and fungi, which act as "Trojan horses" to protect these microorganisms of harsh environmental conditions. In this study, nine Acanthamoeba isolates from bromeliads phylloplane were identified based on the morphology of cyst and trophozoite forms. The genotype level was accessed by the sequence analysis of Acanthamoeba small-subunit rRNA gene. Genotypic characterization grouped five isolates in the genotype T2/T6, three in the T4 genotype and one in the genotype T16. The results obtained indicate that the genotype T2/T6 is common on phylloplane. To predict the pathogenic potential of the Acanthamoeba isolates, thermo and osmotolerance assays were employed, although all isolates were capable of surviving at temperatures of 37°C, other tests will be conducted in the future to determine the potential pathogenic of the isolates. Altogether, our results revealed the importance of the presence of Acanthamoeba associated with bromeliads in Rio Grande do Sul, Brazil, and the necessity for further studies to determine the environmental distribution and the role of these species.

  15. Isolation and molecular characterization of Acanthamoeba genotypes in recreational and domestic water sources from Jamaica, West Indies.

    Science.gov (United States)

    Todd, Cheridah D; Reyes-Batlle, María; Piñero, José E; Martínez-Carretero, Enrique; Valladares, Basilio; Streete, Don; Lorenzo-Morales, Jacob; Lindo, John F

    2015-09-01

    Free living amoebae (FLA) are amphizoic protozoa that are ubiquitous in nature. Infection with FLA may result in neurological, ocular and skin infections. Exposure to Acanthamoeba occurs frequently through water contact and knowledge of the presence of the organisms in water sources is important in understanding transmission dynamics. The distribution of Acanthamoeba was studied in recreational and domestic water samples collected from across Jamaica. Morphological assessment and polymerase chain reaction revealed Acanthamoeba spp. isolates in 50.6% (42/83) and 17.3% (14/81) of recreational and domestic water, respectively. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T3, T4, T5, T10 and T11 corresponding to Acanthamoeba spp: A. griffini, A. triangularis, A. lenticulata, A. culbertsoni and A. hatchetti. Moreover, T4 was the most frequently isolated genotype in both recreational and domestic water. Thermotolerance and osmotolerance assays indicated that most isolates were potentially pathogenic. This is the first report of T3 and T10 genotypes in the Caribbean and the first report of these Acanthamoeba spp. in Jamaican waters. The study shows that there is potential risk of infection to contact wearers who practise poor lens care. Further, Acanthamoeba should be considered as a cause of neurological infections in Jamaica.

  16. The first genotype determination of Acanthamoeba potential threat to human health, isolated from natural water reservoirs in Poland.

    Science.gov (United States)

    Lass, Anna; Szostakowska, Beata; Idzińska, Alicja; Chomicz, Lidia

    2014-07-01

    Different species of amoebae belonging to the genus Acanthamoeba are widely distributed in many parts of the world and known as free-living organisms. Some strains of the protozoans may exist as parasites and cause risk to human health as causative agents of serious human diseases. Currently, in Poland, there is no sufficient information about the distribution of Acanthamoeba strains and their genotypes in the environment. Therefore, 20 environmental surface water samples were collected from different sites located at five water reservoirs in Gdynia, Sopot, and Gdańsk (northern Poland). The material was cultured to obtain Acanthamoeba isolates that were then specifically analyzed with both PCR and real-time PCR assays. Of the 20 samples examined, Acanthamoeba DNA was found in 13 samples tested with the use of real-time PCR; in 10 of them, DNA of the amoeba was also detected using PCR technique. The comparison with sequences available in the GenBank confirmed that the PCR products are fragments of Acanthamoeba 18S rRNA gene and that isolates represent T4 genotype, known as the most common strains related to AK cases. This is the first investigation in Poland describing Acanthamoeba detection in environmental water samples with molecular techniques and genotyping. The results indicate that surface water in Poland may be a source of acanthamoebic strains potentially pathogenic for humans.

  17. Isolation and molecular characterization of Acanthamoeba genotypes isolated from soil sources of public and recreational areas in Iran.

    Science.gov (United States)

    Karamati, Seyed Ahmad; Niyyati, Maryam; Lorenzo-Morales, Jacob; Lasjerdi, Zohreh

    2016-12-01

    Pathogenic strains of Acanthamoeba are causative agents of a sight threating infection of the cornea known as Acanthamoeba keratitis. AK cases have been reported in Iran recently due to inappropriate usage of contact lens maintenance and most patients report a contact with contaminated sources such as dust, water or soil. Sixty soil samples were collected from public and recreational areas in the province of East Azerbaijan, Iran and checked for the presence of Acanthamoeba spp. Samples were cultured on non-nutrient agar plates seeded with heat killed Escherichia coli. PCR and sequencing of the DF3 region were carried out in order to genotype the isolated strains of Acanthamoeba. Thermotolerance and osmotolerance assays were performed in order to investigate the pathogenic potential of isolated Acanthamoeba strains. Acanthamoeba spp. was isolated from 41.6% of soil samples and genotyping of the strains resulted in the identification of genotypes T3, T4, T5 and T11. Most of the isolates belonging to genotypes T3 and T4 showed high pathogenic potential, indicating that they might present a potential health hazard for humans and other animals in this region. To the best of our knowledge, this is the first report on the identification of genotypes T3 and T11 from soil sources in the country.

  18. Acanthamoeba infection in lungs of mice expressed by toll-like receptors (TLR2 and TLR4).

    Science.gov (United States)

    Derda, Monika; Wojtkowiak-Giera, Agnieszka; Kolasa-Wołosiuk, Agnieszka; Kosik-Bogacka, Danuta; Hadaś, Edward; Jagodziński, Paweł P; Wandurska-Nowak, Elżbieta

    2016-06-01

    Toll-like receptors (TLRs) play a key role in the innate immune responses to a variety of pathogens including parasites. TLRs are among the most highly conserved in the evolution of the receptor family, localized mainly on cells of the immune system and on other cells such as lung cells. The aim of this study was to determine for the first time the expression of TLR2 and TLR4 in the lung of Acanthamoeba spp. infected mice using quantitative real-time polymerase chain reaction (Q-PCR) and immunohistochemical (IHC) staining. The Acanthamoeba spp. were isolated from a patient with Acanthamoeba keratitis (AK) (strain Ac 55) and from environmental samples of water from Malta Lake (Poznań, Poland - strain Ac 43). We observed a significantly increased level of expression of TLR2 as well as TLR4 mRNA from 2 to 30 days post Acanthamoeba infection (dpi) in the lungs of mice infected with Ac55 (KP120880) and Ac43 (KP120879) strains. According to our observations, increased TLR2 and TLR4 expression in the pneumocytes, interstitial cells and epithelial cells of the bronchial tree may suggest an important role of these receptors in protective immunity against Acanthamoeba infection in the lung. Moreover, increased levels of TLR2 and TLR4 mRNA expression in infected Acanthamoeba mice may suggest the involvement of these TLRs in the recognition of this amoeba pathogen-associated molecular pattern (PAMP).

  19. Fulminant and fatal encephalitis caused by Acanthamoeba in a kidney transplant recipient: case report and literature review.

    Science.gov (United States)

    Satlin, M J; Graham, J K; Visvesvara, G S; Mena, H; Marks, K M; Saal, S D; Soave, R

    2013-12-01

    Acanthamoeba is the most common cause of granulomatous amebic encephalitis, a typically fatal condition that is classically described as indolent and slowly progressive. We report a case of Acanthamoeba encephalitis in a kidney transplant recipient that progressed to death within 3 days of symptom onset and was diagnosed at autopsy. We also review clinical characteristics, treatments, and outcomes of all published cases of Acanthamoeba encephalitis in solid organ transplant (SOT) recipients. Ten cases were identified, and the infection was fatal in 9 of these cases. In 6 patients, Acanthamoeba presented in a fulminant manner and death occurred within 2 weeks after the onset of neurologic symptoms. These acute presentations are likely related to immunodeficiencies associated with solid organ transplantation that result in an inability to control Acanthamoeba proliferation. Skin lesions may predate neurologic involvement and provide an opportunity for early diagnosis and treatment. Acanthamoeba is an under-recognized cause of encephalitis in SOT recipients and often presents in a fulminant manner in this population. Increased awareness of this disease and its clinical manifestations is essential to attain an early diagnosis and provide the best chance of cure.

  20. Acanthamoeba produces disseminated infection in locusts and traverses the locust blood-brain barrier to invade the central nervous system

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    Kirk Ruth

    2010-07-01

    Full Text Available Abstract Background Many aspects of Acanthamoeba granulomatous encephalitis remain poorly understood, including host susceptibility and chronic colonization which represent important features of the spectrum of host-pathogen interactions. Previous studies have suggested locusts as a tractable model in which to study Acanthamoeba pathogenesis. Here we determined the mode of parasite invasion of the central nervous system (CNS. Results Using Acanthamoeba isolates belonging to the T1 and T4 genotypes, the findings revealed that amoebae induced sickness behaviour in locusts, as evidenced by reduced faecal output and weight loss and, eventually, leading to 100% mortality. Significant degenerative changes of various tissues were observed by histological sectioning. Both isolates produced disseminated infection, with viable amoebae being recovered from various tissues. Histological examination of the CNS showed that Acanthamoeba invaded the locust CNS, and this is associated with disruption of the perineurium cell/glial cell complex, which constitutes the locust blood-brain barrier. Conclusions This is the first study to demonstrate that Acanthamoeba invades locust brain by modulating the integrity of the insect's blood-brain barrier, a finding that is consistent with the human infection. These observations support the idea that locusts provide a tractable model to study Acanthamoeba encephalitis in vivo. In this way the locust model may generate potentially useful leads that can be tested subsequently in mammalian systems, thus replacing the use of vertebrates at an early stage, and reducing the numbers of mammals required overall.

  1. Characterization of patient’s inflammatory pulpal diseases. Caracterización de pacientes con afecciones pulpares inflamatorias

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    Ana Isabel Palenque Guillemí

    2008-06-01

    Full Text Available Background: Pulpal diseases are the most common entities in the dentist emergency consultation. For a correct indication of the treatment, it is very important to establish an accurate diagnosis. Objective: to characterize carrier patients with inflammatory pulpal disease. Methods: a descriptive, prospective cases series study which included 222 patients who came to the dentistry emergency consultation in Area I health policlinic. Age, type of disease, cause, more implicated teeth in the disease, visits, and previous treatments were the variables included. Results: there was no difference at all regarding sex. The most affected group of age was from 35 to 59 years old. The inferior molars were the most affected teeth. The main cause was dental cavities. Obturation was the previous treatment more presented. 46% of the patients attended the dentist in more than one occasion with the same process. Acute irreversible pulpitis was the most predominant pulpal disease. Conclusions: The most common pulpal diseases in our milieu was the acute irreversible pulpitis among the ages 35 and 59 years old.
    Fundamento: Las afecciones pulpares son las enfermedades que más llevan a los pacientes a una consulta de urgencia estomatológica. En estos casos, para una correcta indicación de tratamiento, es de suma importancia el establecimiento de un diagnóstico preciso. Objetivo: Caracterizar pacientes portadores de afectaciones pulpares inflamatorias. Métodos: Estudio descriptivo, prospectivo de serie de casos que incluyó 222 pacientes que asistieron a consulta de urgencia estomatológica en la policlínica del Área 1. Se incluyeron las siguientes variables: edad, tipo de afectación, causa, dientes más afectados, visitas y tratamientos previos. Resultados: No hubo diferencias en cuanto al sexo, el

  2. La peroxidación lipídica en la etiopatogenia de la enfermedad periodontal inflamatoria

    Directory of Open Access Journals (Sweden)

    Bárbara E García Triana

    1998-04-01

    Full Text Available Las especies reactivas del oxígeno están implicadas en la etiopatogenia de la inflamación. Durante la activación de los leucocitos, se liberan grandes cantidades de estas especies, cuya función es la eliminación de los agentes patógenos. Si las defensas antioxidantes de los tejidos no funcionan eficientemente, son inducidas reacciones radicálicas que afectan a las biomoléculas. El ataque a los lípidos de la membrana celular provoca su peroxidación, con la consiguiente formación de nuevas especies radicálicas y metabolitos tóxicos. La presencia de un fuerte infiltrado inflamatorio en los tejidos periodontales, durante la enfermedad periodontal inflamatoria, ha sugerido la posible participación de las especies reactivas del oxígeno en la etiopatogenia de esta enfermedad. Se supone que el ataque de estas sustancias a los tejidos periodontales con deficiente defensa antioxidante, provoca la aparición de la peroxidación lipídica, que puede conducir a la lisis celular y la activación de proteasas.The reative oxygen species are involved in the pathogenesis of inflammation. Leukocyte activation released a great amount of these species the function of which is to eliminate pathogen agents. If antioxidant tissue defenses do not act properly, then radical reactions affecting biomolecules are induced. The attack on cell membrane lipids caused their peroxidation followed by the formation of new radical species and toxic metabolites. The existence of a strong inflammatory infiltrate in periodontal tissues in the course of the periodontal disease suggest the possible involvement of reactive oxygen species in the pathogenesis of this disease. It is supposed that the attack of these substances on the poorly antioxidant defended periodontal tissues makes the lipid peroxidation take place, which may lead to cellular lysis and proteases activation.

  3. Biologic therapies for chronic inflammatory bowel disease Tratamientos biológicos en la enfermedad inflamatoria crónica intestinal

    Directory of Open Access Journals (Sweden)

    M. P. Martínez-Montiel

    2006-04-01

    Full Text Available Crohn's disease (CD and ulcerative colitis (UC make up the so-called chronic inflammatory bowel disease (IBD. Advances in the understanding of IBD pathophysiologic mechanisms in the last few years have allowed the development of novel therapies such as biologic therapies, which at least theoretically represent a more specific management of this disease with fewer side effects. Currently, the only effective and widely accepted biologic therapy for the treatment of intraluminal, fistulizing CD, both for remission induction and maintenance, is infliximab. The role of other monoclonal antibodies such as adalimumab is not clearly established. It could be deemed an alternative for patients with allergic reactions to infliximab, and for those with lost response because of anti-infliximab antibody development. However, relevant issues such as dosage and administration regimen remain to be established. Anti-integrin α4 therapies, despite encouraging results in phase-3 studies, are still unavailable, as their marketing authorization was held back in view of a number of reports regarding progressive multifocal leukoencephalopathy cases. Immunostimulating therapy may be highly relevant in the near future, as it represents a novel strategy against disease with the inclusion of granulocyte-monocyte colony-stimulating factors. Regarding ulcerative colitis, results from the ACT-1 and ACT-2 studies showed that infliximab is also useful for the management of serious UC flare-ups not responding to standard treatment, which will lead to a revision of therapeutic algorithms, where this drug should be given preference before intravenous cyclosporine. In the next few years, the role of anti-CD3 drugs (vilisilizumab, T-cell inhibiting therapies, and epithelial repair and healing stimulating factors will be established.La enfermedad de Crohn (EC y la colitis ulcerosa (CU constituyen la denominada enfermedad inflamatoria crónica intestinal (EII. Los avances producidos

  4. Is safety infliximab during pregnancy in patients with inflammatory bowel disease? ¿El tratamiento con infliximab es seguro durante el embarazo en pacientes con enfermedad inflamatoria intestinal?

    Directory of Open Access Journals (Sweden)

    Federico Argüelles-Arias

    2012-02-01

    foetus. In fact, in our study and in some cases, its withdrawal may lead to a worsening of the disease. However, further control studies are required with larger samples to obtain more representative findings.Introducción: la enfermedad inflamatoria intestinal (EII es un trastorno crónico que debuta en la mayoría de los casos durante la edad reproductiva. Existen pocos datos sobre la seguridad durante el embarazo de los tratamientos disponibles, entre ellos los denominados biológicos, y estos están basados en resultados de casos esporádicos. Objetivos: determinar la seguridad del tratamiento con infliximab (IFX durante el embarazo en mujeres con EII. Un segundo objetivo es observar el efecto que sobre la actividad de la enfermedad tiene el abandono del tratamiento. Material y métodos: se trata de un estudio retrospectivo en el que se incluyeron mujeres con EII embarazadas y que estaban en tratamiento con IFX durante el embarazo. Se incluyeron en el estudio a 5 hospitales de España. La actividad de la enfermedad se midió según el CDAI en la enfermedad de Crohn (EC y la de la colitis ulcerosa (CU según el índice de Truelove-Witts en cada trimestre del embarazo. La edad gestacional, el peso y las enfermedades del feto se determinaron al nacimiento. Resultados: se incluyeron doce mujeres con una edad media de 29 años, 4 diagnosticadas de CU y 8 de EC, con una duración media de la enfermedad de 7 años. Todas salvo una, que se diagnosticó durante el embarazo estaban siendo tratadas con IFX en el momento de la concepción. Seis pacientes recibieron el tratamiento de forma ininterrumpida durante todo el embarazo, 2 suspendieron el tratamiento de forma voluntaria y a tres se les suspendió el tratamiento en el tercer trimestre. Recibieron una dosis media de IFX de 400 mg cada 8 semanas. De las 6 pacientes que recibieron tratamiento continuo, el 50% se mantuvo en remisión. De las pacientes que abandonaron el tratamiento, un 83,3% (todas menos una presentaron un

  5. Distribution of Acanthamoeba Genotypes Isolated from Recreational and Therapeutic Geothermal Water Sources in Southwestern Iran.

    Science.gov (United States)

    Niyyati, Maryam; Saberi, Reza; Latifi, Alireza; Lasjerdi, Zohreh

    2016-01-01

    A comprehensive survey was conducted along 10 km of geothermal rivers in southwestern Iran. A total of 40 water samples were tested for the presence of Acanthamoeba spp., and genotypes were determined by targeting the diagnostic fragment 3 region of the 18S rRNA gene. The pathogenic potential of all positive isolates was also identified using tolerance ability test. High occurrences of Acanthamoeba (50%) were detected in the sampling areas. Based on sequencing analysis, isolates belonging to T4 (93.7%) and T2 (6.25%) genotypes were reported. Thermo- and osmotolerance tests revealed that five strains are highly pathogenic. Since every collection site of this study was associated with high human activity, posting of warning signs, monitoring of recreational water sources, and awareness of high-risk people are of utmost importance. To the best of our knowledge, the present research is the first to report T2 genotype from geothermal water sources in Iran.

  6. [Acanthamoeba keratitis after use of soft contact lenses--case report].

    Science.gov (United States)

    Ziak, P; Ondriska, F; Mrva, M

    2003-09-01

    The case history of a 39-year patient suffering from a deep inflammation of cornea and not responding to conventional antibiotic treatment is presented. The patient was using soft contact lenses during the period of initial symptoms; moreover, he was bathing in thermal bathing pool. A cultivation examination of smears from the area of corneal defect revealed the presence of Acanthamoeba lugdunensis in combination with bacterial infection by Pseudomonas aeruginosa. The available data indicate that it is the first case of acanthamoeba karatitis (AK) after the application of contact lenses in Slovakia. A long-term local treatment with propamidin isethionate (Brolene gtt, ung.) resulted in healing up. The subsequent vision after 16 months since the initial symptoms proved to be 6/12 (0.5). The healing of the centrally localized defect changed the curvature of cornea with consequent hypermetropic shift. The defect completely corrected the patient's myopia (-8.5). The paper describes present possibilities of AK therapy.

  7. An atypical presentation of Acanthamoeba keratitis in a noncontact lens wearer.

    Science.gov (United States)

    Speer, Christine E; Hofmeister, Elizabeth M; Cohen, Elisabeth J

    2003-01-01

    This article presents the case of a 49-year-old man who did not have a history of wearing contact lenses and who developed a rapidly progressive course of Acanthamoeba keratitis. The patient developed stromal keratitis that did not respond to herpes simplex virus therapies. Within 1 week after presentation, the patient progressed from mild anterior stromal haze and edema to a ring infiltrate, epithelial loss, and significant corneal edema. Corneal scrapings demonstrated cysts consistent with Acanthanmoeba keratitis. The patient was admitted to the hospital and placed on intensive medical therapy. He responded to therapy, and at 5 months showed central scarring in a quiet eye. This article presents a case of Acanthamoeba keratitis in a non-contact lens wearer, who was diagnosed clinically and histopathologically within 1 week of onset of symptoms. His case was atypical given his lack of contact lens wear or antecedent trauma and rapid progression to a ring infiltrate, usually seen as late findings.

  8. Superdiffusion dominates intracellular particle motion in the supercrowded space of pathogenic Acanthamoeba castellanii

    CERN Document Server

    Reverey, J F; Bao, H; Leippe, M; Metzler, R; Selhuber-Unkel, C

    2015-01-01

    Acanthamoebae are free-living protists and human pathogens, whose cellular functions and pathogenicity strongly depend on the transport of intracellular vesicles and granules through the cytosol. Using high-speed live cell imaging in combination with single-particle tracking analysis, we show here that the motion of endogenous intracellular particles in the size range from a few hundred nanometers to several micrometers in Acanthamoeba castellanii is strongly superdiffusive and influenced by cell locomotion, cytoskeletal elements, and myosin II. We demonstrate that cell locomotion significantly contributes to intracellular particle motion, but is clearly not the only origin of superdiffusivity. By analyzing the contribution of microtubules, actin, and myosin II motors we show that myosin II is a major driving force of intracellular motion in A. castellanii. The cytoplasm of A. castellanii is supercrowded with intracellular vesicles and granules, such that significant intracellular motion can only be achieved ...

  9. Superdiffusion dominates intracellular particle motion in the supercrowded cytoplasm of pathogenic Acanthamoeba castellanii

    Science.gov (United States)

    Reverey, Julia F.; Jeon, Jae-Hyung; Bao, Han; Leippe, Matthias; Metzler, Ralf; Selhuber-Unkel, Christine

    2015-06-01

    Acanthamoebae are free-living protists and human pathogens, whose cellular functions and pathogenicity strongly depend on the transport of intracellular vesicles and granules through the cytosol. Using high-speed live cell imaging in combination with single-particle tracking analysis, we show here that the motion of endogenous intracellular particles in the size range from a few hundred nanometers to several micrometers in Acanthamoeba castellanii is strongly superdiffusive and influenced by cell locomotion, cytoskeletal elements, and myosin II. We demonstrate that cell locomotion significantly contributes to intracellular particle motion, but is clearly not the only origin of superdiffusivity. By analyzing the contribution of microtubules, actin, and myosin II motors we show that myosin II is a major driving force of intracellular motion in A. castellanii. The cytoplasm of A. castellanii is supercrowded with intracellular vesicles and granules, such that significant intracellular motion can only be achieved by actively driven motion, while purely thermally driven diffusion is negligible.

  10. Bringing forward the new generation of alkoxy-thiourea as potential treatment for Acanthamoeba keratitis

    Science.gov (United States)

    Khairul, Wan M.; Goh, Yit-Peng; Daud, Adibah Izzati; Nakisah, M. A.

    2017-02-01

    Alkoxy substituted thiourea derivatives with general formula of A-ArC(O)NHC(S)NHAr-D which A represents the methoxy group and D denotes -OCnH2n+1 have been successfully synthesised and characterized. In turn, all the synthesised molecules were assayed for anti-amoebic activities towards Acanthamoeba sp to examine the cytotoxicity effect at their IC50 and membrane permeability. As predicted, the findings showed that the synthesised molecules owing promising anti-amoebic activity towards Acanthamoeba sp. To support, the Acridine-orange/Propidium iodide (AOPI) staining result under fluorescence microscopy revealed the treated amoeba cells by these alkoxy thiourea derivatives exhibited loss in their membrane permeability.

  11. Permissivity of insect cells to Waddlia chondrophila, Estrella lausannensis and Parachlamydia acanthamoebae.

    Science.gov (United States)

    Kebbi-Beghdadi, Carole; Fatton, Morgane; Greub, Gilbert

    2015-01-01

    Recent large scale studies questioning the presence of intracellular bacteria of the Chlamydiales order in ticks and fleas revealed that arthropods, similarly to mammals, reptiles, birds or fishes, can be colonized by Chlamydia-related bacteria with a predominant representation of the Rhabdochlamydiaceae and Parachlamydiaceae families. We thus investigated the permissivity of two insect cell lines towards Waddlia chondrophila, Estrella lausannensis and Parachlamydia acanthamoebae, three bacteria representative of three distinct families within the Chlamydiales order, all documented in ticks and/or in other arthropods. We demonstrated that W. chondrophila and E. lausannensis are able to very efficiently multiply in these insect cell lines. E. lausannensis however induced a rapid cytopathic effect, which somehow restricted its replication. P. acanthamoebae was not able to grow in these cell lines even if inclusions containing a few replicating bacteria could occasionally be observed.

  12. Assessment of the efficacy of benzalkonium chloride and sodium hypochlorite against Acanthamoeba polyphaga and Tetrahymena spp.

    Science.gov (United States)

    Vaerewijck, M J M; Sabbe, K; Baré, J; Spengler, H-P; Favoreel, H W; Houf, K

    2012-03-01

    The efficacy of benzalkonium chloride and sodium hypochlorite against Acanthamoeba polyphaga and two Tetrahymena spp. was determined based on the European Standard EN 1276:2009 suspension test. Trophozoite viability was assessed by determination of the membrane integrity using flow cytometry as a fast screening technique. Bovine serum albumin was added to simulate clean (0.3 g/liter) and dirty (3 g/liter) conditions. Benzalkonium chloride caused cell lysis at concentrations above 50 mg/liter under clean and dirty conditions. A concentration of 50 mg of free chlorine per liter had a strong biocidal effect on acanthamoebae and tetrahymenae after 15 min under clean and dirty conditions. Our results suggest that benzalkonium chloride and sodium hypochlorite were effective against the three microorganisms at concentrations commonly applied in the food industry.

  13. Isolation of Acanthamoeba Genotype T4 from a Non-Contact Lens Wearer from the Philippines

    Science.gov (United States)

    Buerano, Corazon C.; Trinidad, Abigail D.; Fajardo, Lindsay Sydney N.; Cua, Irwin Y.; Baclig, Michael O.; Natividad, Filipinas F.

    2014-01-01

    We report the case of a 76-year old Filipino male who presented with pain, redness, and blurring of vision of the right eye. Corneal scraping was done and sent to the St. Luke’s Research and Biotechnology Group for detection and identification of the infectious agent. Morphological detection was performed by allowing the organism from the scraping to grow in 1.5% non-nutrient agar plate with heat-killed E. coli. Trophozoites with acanthopodia and double-walled cysts characteristic of Acanthamoeba were observed within the first and second week of observations, respectively. Molecular identification of the amoebae at the genus level based on the presence of Acanthamoeba-specific amplimer S1, ASA.S1 confirmed the morphological identification. Genotyping through sequence revealed that the organism belonged to T4, which is the genotype commonly present in the eye of keratitis patients. PMID:25589879

  14. Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis.

    Science.gov (United States)

    Peretz, Avi; Geffen, Yuval; Socea, Soergiu D; Pastukh, Nina; Graffi, Shmuel

    2015-08-01

    Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media-non-nutrient agar (NNA), culture on liquid growth media-peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media.

  15. Recent advances in Acanthamoeba keratitis%棘阿米巴角膜炎研究进展

    Institute of Scientific and Technical Information of China (English)

    王小强

    2014-01-01

    Acanthamoeba keratitis is a refractory and progressive eye disease caused by Acanthamoeba.Acanthamoeba exists in natural environment,tap water and corneal contact lens care solution,so the incidence of Acanthamoeba keratitis is increasing because of the corneal contact lens.The diagnosis of Acanthamoeba keratitis depends on the history of disease,signs and symptoms as well as rapidly developed laboratory examinations.The management of Acanthamoeba keratitis includes medical therapy and surgery,and photorefractive keratectomy is a new operation for Acanthamoeba keratitis.Some new knowledge in our understanding of the disease is being produced in recent years.The etiology,novel diagnosis and therapy for this disease were reviewed.%棘阿米巴角膜炎系棘阿米巴原虫感染引起的一种顽固性、进行性角膜炎,为严重的眼部感染性疾病.棘阿米巴原虫大量存在于自来水、角膜接触镜护理液等中,随着角膜接触镜佩戴人数的增多,该病的发生有迅速增加的趋势.棘阿米巴角膜炎的诊断主要依靠病史、临床症状和实验室诊断,尤其是后者在近年来的研究中进展较快.棘阿米巴角膜炎的治疗主要是药物治疗和手术治疗,其中准分子激光角膜切削术已成为一种新的治疗方法.就棘阿米巴角膜炎的病原学、诊断和治疗方面的研究进展进行综述.

  16. Cellular Response of the Amoeba Acanthamoeba castellanii to Chlorine, Chlorine Dioxide, and Monochloramine Treatments ▿

    OpenAIRE

    Mogoa, Emerancienne; Bodet, Charles; Morel, Franck; Rodier, Marie-Hélène; Legube, Bernard; Héchard, Yann

    2011-01-01

    Acanthamoeba castellanii is a free-living amoebae commonly found in water systems. Free-living amoebae might be pathogenic but are also known to bear phagocytosis-resistant bacteria, protecting these bacteria from water treatments. The mode of action of these treatments is poorly understood, particularly on amoebae. It is important to examine the action of these treatments on amoebae in order to improve them. The cellular response to chlorine, chlorine dioxide, and monochloramine was tested o...

  17. Survival and transfer ability of phylogenetically diverse bacterial endosymbionts in environmental Acanthamoeba isolates.

    Science.gov (United States)

    Matsuo, Junji; Kawaguchi, Kouhei; Nakamura, Shinji; Hayashi, Yasuhiro; Yoshida, Mitsutaka; Takahashi, Kaori; Mizutani, Yoshihiko; Yao, Takashi; Yamaguchi, Hiroyuki

    2010-08-01

    Obligate intracellular bacteria are commonly found as endosymbionts of acanthamoebae; however, their survival in and ability to transfer to amoebae are currently uncharacterized. In this study, six bacterial endosymbionts, found in five environmental Acanthamoeba isolates (S13, R18, S23, S31, S40) from different locations of Sapporo city, Japan, were characterized. Phylogenetic analysis revealed that three bacterial endosymbionts (eS23, eS31, eS40a) belonged to α- and β-Proteobacteria phyla and the remaining endosymbionts (eS13, eR18, eS40b) belonged to the order Chlamydiales. The Acanthamoeba isolate (S40) contained two phylogenetically different bacterial endosymbionts (eS40a, eS40b). Fluorescent in situ hybridization analysis showed that all bacterial endosymbionts were diffusely localized within amoebae. Transmission electron microscopy also showed that the endosymbionts were rod-shaped (eS23, eS31, eS40a) or sphere- or crescent-shaped (eS13, eR18, eS40b). No successful culture of these bacteria was achieved using conventional culture methods, but the viability of endosymbionts was confirmed by live/dead staining and RT-PCR methods. However, endosymbionts (except eR18) derived from original host cells lost the ability to be transferred to another Acanthamoebae strains [ATCC strain (C3), environmental strains (S14, R23, S24)]. Thus, our data demonstrate that phylogenetically diverse bacterial endosymbionts found in amoebae maintain a stable interaction with amoebae, but the transferability is limited.

  18. A Rabbit Model of Acanthamoeba Keratitis That Better Reflects the Natural Human Infection.

    Science.gov (United States)

    Feng, Xianmin; Zheng, Wenyu; Wang, Yuehua; Zhao, Donghai; Jiang, Xiaoming; Lv, Shijie

    2015-08-01

    Acanthamoeba species are ubiquitous, free-living protozoa that can invade the cornea and result in Acanthamoeba keratitis (AK), a painful progressive sight-threatening corneal disease. Disease progression in current animal models is too rapid to mimic AK in humans accurately. This study provides a novel method for establishing AK in rabbits and compared it with the conventional method with regard to pathogenesis and immune response in humans. The New Zealand white rabbits were randomly divided into two experimental groups (Groups A and B). Rabbits in the Group A (n = 14) received intrastromal injections of 1 × 10(4) /100 µL Acanthamoeba healyi trophozoites (conventional AK model). The Group B animals (n = 14) received microinjections of 1 × 10(4) /10 µL A. healyi trophozoites between the corneal epithelium and Bowman's layer, anterior to the corneal stroma (novel AK model). In addition, two rabbits were left untreated as normal controls. AK in the treated rabbits was evaluated clinically, histopathologically, and immunologically for 35 days. AK was successfully established in both the conventional and novel model groups. Compared with the Group A, AK in the Group B displayed an efficient immune response with less severe pathology. Moreover, the self-limiting but chronic nature of the infection in the Group B was strikingly similar to that of AK in humans. The novel animal model for AK described here more closely simulates the pathogenesis and immune response of Acanthamoeba corneal infection in humans than the animal models currently in use.

  19. A novel antiamoebic agent against Acanthamoeba sp. - A causative agent for eye keratitis infection

    Science.gov (United States)

    Kusrini, Eny; Hashim, Fatimah; Azmi, Wan Nor Nadhirah Wan Noor; Amin, Nakisah Mat; Estuningtyas, Ari

    2016-01-01

    The terbium trinitrate.trihydrate.18-crown ether-6, Tb(NO3)3(OH2)3.(18C6) complex has been characterized by elemental analysis, photoluminescence and single X-ray diffraction. The IC50 values were determined based on MTT assay while light and fluorescence microscopy imaging were employed to evaluate the cellular morphological changes. Alkaline comet assay was performed to analyze the DNA damage. The photoluminescence spectrum of the Tb complex excited at 325 nm displayed seven luminescence peaks corresponding to the 5D4 → 7F0, 1, 2, 3, 4, 5, 6 transitions. The cytotoxicity and genotoxicity studies indicated that the Tb(NO3)3(OH2)3.(18C6) complex and its salt form as well as the 18C6 molecule have excellent anti-amoebic activity with very low IC50 values are 7, 2.6 and 1.2 μg/mL, respectively, with significant decrease (p Acanthamoeba viability when the concentration was increased from 0 to 30 μg/mL. The mode of cell death in Acanthamoeba cells following treatment with the Tb complex was apoptosis. This is in contrast to the Tb(NO3)3.6H2O salt- and 18C6 molecule-treated Acanthamoeba, which exhibited necrotic type cells. The percentage of DNA damage following treatment with all the compounds at the IC25 values showed high percentage of type 1 with the % nuclei damage are 14.15 ± 2.4; 46.00 ± 4.2; 36.36 ± 2.4; 45.16 ± 0.6%, respectively for untreated, treated with Tb complex, Tb salt and 18C6 molecule. The work features promising potential of Tb(NO3)3(OH2)3.(18C6) complex as anti-amoebic agent, representing a therapeutic option for Acanthamoeba keratitis infection.

  20. A novel antiamoebic agent against Acanthamoeba sp.--A causative agent for eye keratitis infection.

    Science.gov (United States)

    Kusrini, Eny; Hashim, Fatimah; Azmi, Wan Nor Nadhirah Wan Noor; Amin, Nakisah Mat; Estuningtyas, Ari

    2016-01-15

    The terbium trinitrate.trihydrate.18-crown ether-6, Tb(NO3)3(OH2)3.(18C6) complex has been characterized by elemental analysis, photoluminescence and single X-ray diffraction. The IC50 values were determined based on MTT assay while light and fluorescence microscopy imaging were employed to evaluate the cellular morphological changes. Alkaline comet assay was performed to analyze the DNA damage. The photoluminescence spectrum of the Tb complex excited at 325 nm displayed seven luminescence peaks corresponding to the (5)D4→(7)F(0, 1, 2, 3, 4, 5, 6) transitions. The cytotoxicity and genotoxicity studies indicated that the Tb(NO3)3(OH2)3.(18C6) complex and its salt form as well as the 18C6 molecule have excellent anti-amoebic activity with very low IC50 values are 7, 2.6 and 1.2 μg/mL, respectively, with significant decrease (pAcanthamoeba viability when the concentration was increased from 0 to 30 μg/mL. The mode of cell death in Acanthamoeba cells following treatment with the Tb complex was apoptosis. This is in contrast to the Tb(NO3)3.6H2O salt- and 18C6 molecule-treated Acanthamoeba, which exhibited necrotic type cells. The percentage of DNA damage following treatment with all the compounds at the IC25 values showed high percentage of type 1 with the % nuclei damage are 14.15±2.4; 46.00±4.2; 36.36±2.4; 45.16±0.6%, respectively for untreated, treated with Tb complex, Tb salt and 18C6 molecule. The work features promising potential of Tb(NO3)3(OH2)3.(18C6) complex as anti-amoebic agent, representing a therapeutic option for Acanthamoeba keratitis infection.

  1. Identification and properties of proteases from an Acanthamoeba isolate capable of producing granulomatous encephalitis

    Directory of Open Access Journals (Sweden)

    Jarroll Edward L

    2006-05-01

    Full Text Available Abstract Background Granulomatous amoebic encephalitis due to Acanthamoeba is often a fatal human disease. However, the pathogenesis and pathophysiology of Acanthamoeba encephalitis remain unclear. In this study, the role of extracellular Acanthamoeba proteases in central nervous system pathogenesis and pathophysiology was examined. Results Using an encephalitis isolate belonging to T1 genotype, we observed two major proteases with approximate molecular weights of 150 KD and 130 KD on SDS-PAGE gels using gelatin as substrate. The 130 KD protease was inhibited with phenylmethylsulfonyl fluoride (PMSF suggesting that it is a serine protease, while the 150 KD protease was inhibited with 1, 10-phenanthroline suggesting that it is a metalloprotease. Both proteases exhibited maximal activity at neutral pH and over a range of temperatures, indicating their physiological relevance. These proteases degrade extracellular matrix (ECM, which provide structural and functional support to the brain tissue, as shown by the degradation of collagen I and III (major components of collagenous ECM, elastin (elastic fibrils of ECM, plasminogen (involved in proteolytic degradation of ECM, as well as casein and haemoglobin. The proteases were purified partially using ion-exchange chromatography and their effects were tested in an in vitro model of the blood-brain barrier using human brain microvascular endothelial cells (HBMEC. Neither the serine nor the metalloprotease exhibited HBMEC cytotoxicity. However, the serine protease exhibited HBMEC monolayer disruptions (trypsin-like suggesting a role in blood-brain barrier perturbations. Conclusion Overall, these data suggest that Acanthamoeba proteases digest ECM, which may play crucial role(s in invasion of the brain tissue by amoebae.

  2. Susceptibility of Aedes aegypti (Diptera: Culicidae) to Acanthamoeba polyphaga (Sarcomastigophora: Acanthamoebidae).

    Science.gov (United States)

    Rott, Marilise; Caumo, Karin; Sauter, Ismael; Eckert, Janina; da Rosa, Luana; da Silva, Onilda

    2010-06-01

    To date there is no report on mosquitoes infected with free-living amoebae. For this reason, the aim of this study was to verify if Aedes aegypti could be susceptible to Acanthamoeba polyphaga under laboratory conditions, so trophozoites were offered as a unique food resource for larvae of first instar. The results show that those amoebae are able to infect and colonize the mosquito gut and could be re-isolated of all stages of the mosquito (larvae, pupae, and adults).

  3. Isolation of Acanthamoeba Spp. from Drinking Waters in Several Hospitals of Iran

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    HR Bagheri

    2010-06-01

    Full Text Available Background: Acanthamoeba is an opportunistic amphizoic protozoan found in different wa­ter sources including swimming pool as well as in sewage. The aim of this study was to in­vestigate the prevalence of Acanthamoeba in tap-water samples in Iran.Method: In this descriptive cross-sectional study, 94 samples of cold and warm tap-water were collected from different wards of hospitals in 13 cities of Iran in 2007-2008. Free resi­dual chlorine, pH, and temperature of samples were measured. After filtration through multi­pore nylon membrane, samples were cultured on non-nutrient agar. Then we investigated ex­istence of Acanthamoeba by reverse contrast phase microscope.Results: Acanthamoeba was found in 45 samples (48%. Thirty-four and 11 positive samples were collected from cold and warm tap water, respectively. The samples belonged to the cate­gory of 20-30 °C temperature with 0-2 ppm free residual chlorine and pH 6-7.4 showed the most coincidence to the positive cases. The greatest proportion of positive samples was ob­tained from Mashhad hospitals, while all samples collected from Arak and Semnan hospitals were negative.Conclusion: considering the results of this study and the pathogenic role of this protozoan on pa­tients with immunodeficiency, as well as capability of this microorganism in carrying other pathogens such as Legionella, further studies are needed. What is more important, potable water in hospitals should follow the procedure of treatment and sanitation, in order to prevent the relevant nosocomial infections.

  4. Acanthamoeba meningoencephalitis in immunocompetent: A case report and review of literature.

    Science.gov (United States)

    Khanna, Vinay; Shastri, Ba; Anusha, G; Mukhopadhayay, Chiranjay; Khanna, Ruchee

    2014-07-01

    A 30-year-old manual laborer from Karnataka, India presented with intermittent low grade fever and diffuse headache for 1 month. On examination, patient had enlarged supraclavicular and cervical lymph nodes. Patient had positive Kernig's sign and neck stiffness. Motor, sensory and cranial nerve examinations were within the normal limits. Abdominal, cardiovascular and chest examination did not yield any positive findings. Contrast enhanced computed tomography head was normal. Patient was suspected to have extrapulmonary tuberculosis. Patient was started on antitubercular drugs. Diagnostic lumbar puncture was performed. Wet mount and Giemsa smear preparation of cerebrospinal fluid (CSF) showed trophozoites suggestive of Acanthamoeba. CSF was cultured onto non-nutrient agar with an overlay of Escherichia coli. Wet mount made from the culture media yielded cysts and trophozoites of Acanthamoeba spp. Patient was diagnosed with Acanthamoeba meningitis and was started on specific therapy with Rifampicin 600 mg once a day, Cotrimoxazole 960 mg twice-a-day and Fluconazole 400 mg once daily for 2 weeks. Patient had a complete recovery and was discharged from the hospital.

  5. Caspase-like proteins: Acanthamoeba castellanii metacaspase and Dictyostelium discoideum paracaspase, what are their functions?

    Indian Academy of Sciences (India)

    Entsar Saheb; Wendy Trzyna; John Bush

    2014-12-01

    Caspases are cysteine proteases that are important regulators of programmed cell death in animals. Two novel relatives to members of the caspase families metacaspases and paracaspase have been discovered. Metacaspase type-1 was identified in Acanthamoeba castellanii, an opportunistic protozoan parasite that causes severe diseases in humans. Paracaspase was found in the non-pathogenic protozoan Dictyostelium discoideum. Since their discovery in Acanthamoeba and Dictyostelium, metacaspases and paracaspases have remained poorly characterized. At present we do not have sufficient data about the molecular function of these caspase-like proteins or their role, if any, in programmed cell death. How these caspase proteins function at the molecular level is an important area of study that will provide insight into their potential for treatment therapies against Acanthamoeba infection and other similar parasitic protozoan. Additionally, finding the molecular functions of these caspase-like proteins will provide information concerning their role in more complex organisms.The aim of this article was to review recent discoveries about metacaspases and paracaspases as regulators of apoptotic and non-apoptotic processes.

  6. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii trophozoites and cysts.

    Science.gov (United States)

    Matin, Abdul; Jung, Suk-Yul

    2011-12-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. coli K12 was also applied. The association ratio of E. coli K1 with A. castellanii was 4.3 cfu per amoeba for 1 hr but E. coli K5 with A. castellanii was 1 cfu per amoeba for 1 hr. By invasion and survival assays, E. coli K5 was recovered less than E. coli K1 but still alive inside A. castellanii. E. coli K1 and K5 survived and multiplied intracellularly in A. castellanii. The survival assay was performed under a favourable condition for 22 hr and 43 hr with the encystment of A. castellanii. Under the favourable condition for the transformation of trophozoites into cysts, E. coli K5 multiplied significantly. Moreover, the pathogenic potential of E. coli K1 from A. castellanii cysts exhibited no changes as compared with E. coli K1 from A. castellanii trophozoites. E. coli K5 was multiplied in A. castellanii trophozoites and survived in A. castellanii cysts. Therefore, this study suggests that E. coli K5 can use A. castellanii as a reservoir host or a vector for the bacterial transmission.

  7. Expression levels of encystation mediating factors in fresh strain of Acanthamoeba castellanii cyst ESTs.

    Science.gov (United States)

    Moon, Eun-Kyung; Chung, Dong-Il; Hong, Yeonchul; Kong, Hyun-Hee

    2011-04-01

    The life cycle of Acanthamoeba consists of two stages, trophozoite and cyst. The cyst form is resistant to almost all antibiotics. By long term cultivation, Acanthamoeba severely attenuated the encysting ability. To determine the changing of gene expression by the long term cultivation, especially focusing an encystation mediating factors, this study compared the ESTs of the fresh strain and the old strain, and trophozoite. Comparison of the KOG (euKaryotic Orthologous Groups) analysis relative to trophozoite revealed higher percentages of cyst ESTs related to G (Carbohydrate transport and metabolism), H (Coenzyme transport and metabolism), I (Lipid transport and metabolism), D (Cell cycle control, cell division, chromosome partitioning), T (signal transduction mechanisms), and O (Posttranslational modification, protein turnover, chaperones). In addition to this result, KOG analysis of fresh strain relative to old strain showed higher percentage of cyst ESTs related to metabolism category and T (signal transduction mechanisms) article. ESTs of the fresh strain revealed more various gene profiles compared to the old strain including encystation mediating factors like autophagy related proteins (Z article) and signal transduction proteins (T article). Twenty seven kinds of protein kinase C (PKC) like genes were detected in cyst or trophozoite ESTs and twenty one of them were highly expressed during encystation. The information of the expressed genes during encystation in only the fresh strain will provide new clues to understanding the encystation mechanism of encysting protozoa including Acanthamoeba.

  8. In vitro efficacy of corifungin against Acanthamoeba castellanii trophozoites and cysts.

    Science.gov (United States)

    Debnath, Anjan; Tunac, Josefino B; Silva-Olivares, Angélica; Galindo-Gómez, Silvia; Shibayama, Mineko; McKerrow, James H

    2014-01-01

    Painful blinding keratitis and fatal granulomatous amebic encephalitis are caused by the free-living amebae Acanthamoeba spp. Several prescription eye medications are used to treat Acanthamoeba keratitis, but the infection can be difficult to control because of recurrence of infection. For the treatment of encephalitis, no single drug was found useful, and in spite of the use of a combination of multiple drugs, the mortality rate remains high. Therefore, efficient, novel drugs are urgently needed for the treatment of amebic keratitis and granulomatous amebic encephalitis. In this study, we identified corifungin, a water-soluble polyene macrolide, as amebicidal. In vitro, it was effective against both the trophozoites and the cysts. Transmission electron microscopy of Acanthamoeba castellanii incubated with corifungin showed the presence of swollen mitochondria, electron-dense granules, degeneration of cytoplasm architecture, and loss of nuclear chromatin structure. These changes were followed by lysis of amebae. Corifungin also induced the encystment process of A. castellanii. There were alterations in the cyst cell wall followed by lysis of the cysts. Corifungin is a promising therapeutic option for keratitis and granulomatous amebic encephalitis.

  9. In vitro effects of selected contact lens care solutions on Acanthamoeba castellanii strains in Poland.

    Science.gov (United States)

    Padzik, Marcin; Chomicz, Lidia; Szaflik, Jacek P; Chruścikowska, Agnieszka; Perkowski, Konrad; Szaflik, Jerzy

    2014-11-01

    Free-living, cosmopolitan amoebae of the Acanthamoeba genus may be the causative agents of Acanthamoeba keratitis (AK) - a progressive, vision-threatening infection of the human cornea described particularly among contact lens wearers. Use of contact lens care solutions, effective against these organisms, is important in preventing AK infections. 3 different strains of Acanthamoeba castellanii of the T4 genotype (Neff strain and two others, isolated from patients with AK) were exposed to 4 selected multipurpose contact lens care solutions available in Poland: Ciba Vision AoSept Plus, Bausch & Lomb ReNu MultiPlus, Alcon Opti-Free, Ciba Vision Solo Care Aqua. No amoebicidal effect was observed. The strongest amoebostatic effect was visible after 24h of exposition to Opti-Free and ReNu solution and associated with percentage increase of rounded, motionless forms. This is significantly longer than minimum disinfection time recommended by manufacturers of all tested multipurpose solutions. Surprisingly, no clear induction of the encystation process was observed.

  10. Biochemical and cellular mechanisms regulating Acanthamoeba castellanii adherence to host cells.

    Science.gov (United States)

    Soto-Arredondo, K J; Flores-Villavicencio, L L; Serrano-Luna, J J; Shibayama, M; Sabanero-López, M

    2014-04-01

    Free-living amoebae belonging to the genus Acanthamoeba are the causative agents of infections such as amoebic keratitis (AK), granulomatous amoebic encephalitis (GAE) and cutaneous lesions. The mechanisms involved in the establishment of infection are unknown. However, it is accepted that the initial phase of pathogenesis involves adherence to the host tissue. In this work, we analysed surface molecules with an affinity for epithelial and neuronal cells from the trophozoites of Acanthamoeba castellanii. We also investigated the cellular mechanisms that govern the process of trophozoite adhesion to the host cells. We first used confocal and epifluorescence microscopy to examine the distribution of the A. castellanii actin cytoskeleton during interaction with the host cells. The use of drugs, as cytochalasin B (CB) and latrunculin B (LB), revealed the participation of cytoskeletal filaments in the adhesion process. In addition, to identify the proteins and glycoproteins on the surface of A. castellanii, the trophozoites were labelled with biotin and biotinylated lectins. The results revealed bands of surface proteins, some of which were glycoproteins with mannose and N-acetylglucosamine residues. Interaction assays of biotinylated amoebae proteins with epithelial and neuronal cells showed that some surface proteins had affinity for both cell types. The results of this study provide insight into the biochemical and cellular mechanisms of the Acanthamoeba infection process.

  11. Anaerobic respiration: In vitro efficacy of Nitazoxanide against mitochondriate Acanthamoeba castellanii of the T4 genotype.

    Science.gov (United States)

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Farooq, Maria; Khan, Naveed Ahmed

    2015-10-01

    Acanthamoeba is an opportunistic protist pathogen that is responsible for serious human and animal infection. Being one of the most frequently isolated protists from the environment, it is likely that it readily encounters microaerophilic environments. For respiration under anaerobic or low oxygen conditions in several amitochondriate protists, decarboxylation of pyruvate is catalyzed by pyruvate ferredoxin oxidoreductase instead of pyruvate dehydrogenase. In support, Nitazoxanide, an inhibitor of pyruvate ferredoxin oxidoreductase, is effective and non-mutagenic clinically against a range of amitochondriate protists, Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. The overall aim of the present study was to determine in vitro efficacy of Nitazoxanide against Acanthamoeba castellanii. At micromolar concentrations, the findings revealed that Nitazoxanide neither affected A. castellanii growth or viability nor amoeba-mediated host cell monolayer damage in vitro or extracellular proteolytic activities. Similarly, microaerophilic conditions alone had no significant effects. In contrast, microaerophilic conditions together with Nitazoxanide showed amoebicidal effects and inhibited A. castellanii-mediated host cell monolayer damage as well as extracellular proteases. Using encystation assays, it was observed that Nitazoxanide inhibited trophozoite transformation into cysts both under aerophilic and microaerophilic conditions. Furthermore, pre-treatment of cysts with Nitazoxanide inhibited A. castellanii excystation. These findings are important in the identification of potential targets that could be useful against parasite-specific respiration as well as to understand the basic biology of the life cycle of Acanthamoeba.

  12. Photodynamic inactivation of Acanthamoeba polyphaga with curcuminoids: an in vitro study

    Science.gov (United States)

    Corrêa, Thaila Q.; Geralde, Mariana C.; Carvalho, Mariana T.; Bagnato, Vanderlei S.; Kurachi, Cristina; de Souza, Clovis W. O.

    2016-03-01

    Acanthamoeba polyphaga are free-living amoebae that can be considered potentially pathogenic organisms by cause serious human infections, including keratitis and granulomatous amoebic encephalitis that usually results in death. Photodynamic inactivation (PDI) has been used for the biological control of microorganisms and can be promise in the control of Acanthamoeba infections. This study evaluated the in vitro effectiveness of PDI in A. polyphaga using curcuminoids salt as photosensitizer (PS) besides observing morphological changes caused by this PS in this organism, in confocal microscopy. A. polyphaga trophozoites were grown at 37°C in PYG medium for 48 to 72 hours. After, the trophozoites were incubated with PS solution during one hour and the samples were irradiated using light-emitting diodes at 460 nm at light doses 30 and 50 J/cm2. The results revealed reduction of 27.7%, 61.4% and 82.5% at 30 J/cm2 and 75.2%, 85.0% and 95.9% at 50 J/cm2, respectively, at curcuminoid salt concentrations of 500, 1000 and 1500 μg/mL. Through fluorescence images, it was possible to visualize the curcuminoid salt's uptake by the trophozoites. The PS showed toxicity to amoebae, in the dark, but the irradiation in PDI contributed to amoebae death effect. These data suggest that PDI may be an application of therapeutic intervention against Acanthamoeba infections, since it was effective in the inactivation of these amoebae.

  13. Presumed late recurrence of Acanthamoeba keratitis exacerbated by exposure to topical corticosteroids

    Directory of Open Access Journals (Sweden)

    Dipika V Patel

    2013-01-01

    Full Text Available A 28-year-old female with a history of contact lens wear presented with a 1 week history of pain and photophobia in her left eye. In vivo confocal microscopy (IVCM and corneal scrape confirmed the diagnosis of Acanthamoeba keratitis (AK which was treated with intensive topical propamidine isethionate (0.1% and chlorhexidine (0.02% with tapering dosage over 11 months. Five years after complete resolution of AK and cessation of all contact lens wear, the subject presented to her optometrist with a history of ocular discomfort and mild photophobia. Without further investigation she was prescribed topical corticosteroids. Three weeks later she presented with pain and reduced vision in the left eye. Slit-lamp examination revealed focal, inferior corneal stromal edema. IVCM confirmed widespread Acanthamoeba cysts. Treatment with topical polyhexamethylene biguanide (PHMB 0.02% and propamidine isethionate 0.1% resulted in resolution of the AK. Despite an initially mild AK, this subject presumably retained viable Acanthamoeba cysts in her cornea 5 years after the initial episode. This report highlights the importance of caution when using corticosteroids in patients with a previous history of AK, even in the relatively distant past. Patients with AK should be warned regarding the risks of recurrence following presumed resolution.

  14. Acanthamoeba T4, T5 and T11 isolated from mineral water bottles in southern Brazil.

    Science.gov (United States)

    Maschio, Vinicius José; Chies, Fernanda; Carlesso, Ana Maris; Carvalho, Amanda; Rosa, Sayonara Peixoto; Van Der Sand, Sueli Teresinha; Rott, Marilise Brittes

    2015-01-01

    Acanthamoeba is a protist potential pathogen, capable of causing a blinding keratitis in contact lens wearers and disseminated infection, leading to granulomatous amebic encephalitis in immunocompromised individuals. This amoeba is a ubiquitous organism that has been isolated from various domestic water systems, such as cooling towers and hospital water networks. The objective of this work was to investigate the presence of Acanthamoeba in mineral water bottles marketed in Porto Alegre, southern Brazil. Positive samples were further classified at the genotype level after sequencing the ASA.S1 region of 18S rDNA gene. Six of the eight isolates belonged to T5 genotype, one to T4 genotype, and one was T11. Several genotypes have been reported worldwide as causative of pathologies in humans, including genotypes T4, T5 and T11. Overall, the widespread distribution of potentially pathogenic Acanthamoeba strains in the studied source demands more awareness within the public and health professionals, because this pathogen is emerging as a risk for human health worldwide.

  15. Inactivation of Acanthamoeba spp. and Other Ocular Pathogens by Application of Cold Atmospheric Gas Plasma

    Science.gov (United States)

    Shama, Gilbert; Andrew, Peter W.

    2016-01-01

    Currently there are estimated to be approximately 3.7 million contact lens wearers in the United Kingdom and 39.2 million in North America. Contact lens wear is a major risk factor for developing an infection of the cornea known as keratitis due to poor lens hygiene practices. While there is an international standard for testing disinfection methods against bacteria and fungi (ISO 14729), no such guidelines exist for the protozoan Acanthamoeba, which causes a potentially blinding keratitis most commonly seen in contact lens wearers, and as a result, many commercially available disinfecting solutions show incomplete disinfection after 6 and 24 h of exposure. Challenge test assays based on international standard ISO 14729 were used to determine the antimicrobial activity of cold atmospheric gas plasma (CAP) against Pseudomonas aeruginosa, Candida albicans, and trophozoites and cysts of Acanthamoeba polyphaga and Acanthamoeba castellanii. P. aeruginosa and C. albicans were completely inactivated in 0.5 min and 2 min, respectively, and trophozoites of A. polyphaga and A. castellanii were completely inactivated in 1 min and 2 min, respectively. Furthermore, for the highly resistant cyst stage of both species, complete inactivation was achieved after 4 min of exposure to CAP. This study demonstrates that the CAP technology is highly effective against bacterial, fungal, and protozoan pathogens. The further development of this technology has enormous potential, as this approach is able to deliver the complete inactivation of ocular pathogens in minutes, in contrast to commercial multipurpose disinfecting solutions that require a minimum of 6 h. PMID:26994079

  16. Inactivation of Acanthamoeba spp. and Other Ocular Pathogens by Application of Cold Atmospheric Gas Plasma.

    Science.gov (United States)

    Heaselgrave, Wayne; Shama, Gilbert; Andrew, Peter W; Kong, Michael G

    2016-05-15

    Currently there are estimated to be approximately 3.7 million contact lens wearers in the United Kingdom and 39.2 million in North America. Contact lens wear is a major risk factor for developing an infection of the cornea known as keratitis due to poor lens hygiene practices. While there is an international standard for testing disinfection methods against bacteria and fungi (ISO 14729), no such guidelines exist for the protozoan Acanthamoeba, which causes a potentially blinding keratitis most commonly seen in contact lens wearers, and as a result, many commercially available disinfecting solutions show incomplete disinfection after 6 and 24 h of exposure. Challenge test assays based on international standard ISO 14729 were used to determine the antimicrobial activity of cold atmospheric gas plasma (CAP) against Pseudomonas aeruginosa, Candida albicans, and trophozoites and cysts of Acanthamoeba polyphaga and Acanthamoeba castellanii P. aeruginosa and C. albicans were completely inactivated in 0.5 min and 2 min, respectively, and trophozoites of A. polyphaga and A. castellanii were completely inactivated in 1 min and 2 min, respectively. Furthermore, for the highly resistant cyst stage of both species, complete inactivation was achieved after 4 min of exposure to CAP. This study demonstrates that the CAP technology is highly effective against bacterial, fungal, and protozoan pathogens. The further development of this technology has enormous potential, as this approach is able to deliver the complete inactivation of ocular pathogens in minutes, in contrast to commercial multipurpose disinfecting solutions that require a minimum of 6 h.

  17. Detection of Acanthamoeba on the ocular surface in a Spanish population using the Schirmer strip test: pathogenic potential, molecular classification and evaluation of the sensitivity to chlorhexidine and voriconazole of the isolated Acanthamoeba strains.

    Science.gov (United States)

    Rocha-Cabrera, Pedro; Reyes-Batlle, María; Martín-Navarro, Carmen María; Dorta-Gorrín, Alexis; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Piñero, José E; Martín-Barrera, Fernando; Valladares, Basilio; Lorenzo-Morales, Jacob

    2015-08-01

    Pathogenic strains of Acanthamoeba are causative agents of a sight-threatening infection of the cornea known as Acanthamoeba keratitis, which is often associated with the misuse of contact lenses. However, there is still a question remaining to be answered, which is whether these micro-organisms are present on the ocular surface of healthy individuals. Therefore, the aim of this study was to determine the presence of Acanthamoeba on the ocular surface in healthy patients and also in those with other ocular surface infections. Sterile Schirmer test strips were used to collect samples from a group of patients who attended an ophthalmology consultation at the Hospital del Norte, Icod de los Vinos, Tenerife, Canary Islands. Most of the patients (46 individuals, 79.31  %) presented ocular surface pathologies such as blepharitis or conjunctivitis; the rest did not present any pathology. None of the patients included in the study wore contact lenses. The collected samples were cultured in 2  % non-nutrient agar plates and positive plates were then cultured in axenic conditions for further analyses. Molecular analysis classified all isolated strains as belonging to Acanthamoeba genotype tbl4, and osmotolerance and thermotolerance assays revealed that all strains were potentially pathogenic. Furthermore, all strains were assayed for sensitivity against voriconazole and chlorhexidine. Assays showed that both drugs were active against the tested strains. In conclusion, the Schirmer strip test is proposed as an effective tool for the detection of Acanthamoeba on the ocular surface.

  18. Chronic Inflammatory Periodontal Disease in Patients with Human Immunodeficiency Virus. Enfermedad periodontal inflamatoria crónica en pacientes infectados con el virus de inmunodeficiencia humana.

    Directory of Open Access Journals (Sweden)

    Iralys Benítez Guzmán

    2009-07-01

    Full Text Available Background: The Chronic Inflammatory Periodontal Disease is related with multiple risk factors. Those patients with human immunodeficiency virus have higher risk of presenting this disease and it is usually more serious in these cases. Objective: To describe the prevalence of Chronic Inflammatory Periodontal Disease in patients with HIV. Methods: Descriptive, observational, cross-sectional study including patients with HIV in Sancti Spiritus province. The occurrence of the disease was determined after the Periodontics Cuban Standards, and oral hygiene was assessed through the simplified oral hygiene index. Other variables were measured, such as smoking habits, T CD4+ lymphocyte counting and virus load. The independent association of each risk factor with the disease was determined through a logistic regression model. Results: The 56, 5 % of the 154 patients presented Chronic Inflammatory Periodontal Disease; 60 (39.0% gingivitis and 27 (17,5% periodontitis. Gingivitis was associated with poor oral hygiene (OR: 3,71 and periodontitis with smoking habit (OR: 5,20. The severe forms of periodontitis occurred mainly in patients with lymphocyte counting lower than 500 cells/mm3 . Conclusions: The prevalence of Chronic Inflammatory Periodontal Disease in patients with HIV in Sancti Spiritus province is linked to known risk factors such as smoking habits and oral hygiene.Fundamento: La enfermedad periodontal inflamatoria crónica es un trastorno relacionado con diversidad de factores de riesgo. Los pacientes infectados con el virus de inmunodeficiencia humana tienen mayor riesgo para padecerla y en ellos muchas veces se agrava.  Objetivo: Describir la prevalencia de la enfermedad periodontal inflamatoria crónica en pacientes infectados con el virus de inmunodeficiencia humana. Métodos: Estudio observacional

  19. Early expression of the type III secretion system of Parachlamydia acanthamoebae during a replicative cycle within its natural host cell Acanthamoeba castellanii.

    Science.gov (United States)

    Croxatto, Antony; Murset, Valérie; Chassot, Bérénice; Greub, Gilbert

    2013-12-01

    The type three secretion system (T3SS) operons of Chlamydiales bacteria are distributed in different clusters along their chromosomes and are conserved at both the level of sequence and genetic organization. A complete characterization of the temporal expression of multiple T3SS components at the transcriptional and protein levels has been performed in Parachlamydia acanthamoebae, replicating in its natural host cell Acanthamoeba castellanii. The T3SS components were classified in four different temporal clusters depending on their pattern of expression during the early, mid- and late phases of the infectious cycle. The putative T3SS transcription units predicted in Parachlamydia are similar to those described in Chlamydia trachomatis, suggesting that T3SS units of transcriptional expression are highly conserved among Chlamydiales bacteria. The maximal expression and activation of the T3SS of Parachlamydia occurred during the early to mid-phase of the infectious cycle corresponding to a critical phase during which the intracellular bacterium has (1) to evade and/or block the lytic pathway of the amoeba, (2) to differentiate from elementary bodies (EBs) to reticulate bodies (RBs), and (3) to modulate the maturation of its vacuole to create a replicative niche able to sustain efficient bacterial growth.

  20. Molecular Characterization and Sequencing of a Gene Encoding Mannose Binding Protein in an Iranian Isolate of Acanthamoeba castellanii as a Major Agent of Acanthamoeba Keratitis

    Directory of Open Access Journals (Sweden)

    SH Farnia

    2008-07-01

    Full Text Available Background: Acanthamoeba castellanii is the important cause of amoebic keratitis in Iran. The key molecule in pathogenesis of Acanthamoeba keratitis is Mannose Binding Protein (MBP led to adhesion of amoeba to corneal epithelium. Subsequent to adhesion other cytopathic effects occur. The goal of this study was to identify the molecular characterization of a gene encoding MBP in an Iranian isolate of A.castellanii in order to pave the way for further investigations such as new therapeutic advances or immunization. Methods: A.castellanii was cultured on non nutrient agar. Extraction of DNA was performed by phenol-chloroform method. After designing a pair of primer for the gene encoding MBP, PCR analysis was performed. Finally, the PCR product has been sequenced and the result submitted to the gene data banks. Results: An MBP gene of 1081 nucleotides was sequenced. This fragment contained three introns and encodes a protein with 194 amino acids. Homology search by Blast program showed a significant homology with the MBP gene in gene data banks (96%. Besides, the identity of amino acids with the other MBPs in gene data banks was about 86%. Conclusion: We isolated and sequenced a gene fragment encoding MBP in an Iranian isolate of A.castellanii. Molecular characterization of this important gene is the first step in pursuing researches such as developing better therapeutic agents, immunization of population at risk or even developing a diagnostic tool by PCR techniques.

  1. Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain.

    Science.gov (United States)

    Huang, Jian-Ming; Lin, Wei-Chen; Li, Sung-Chou; Shih, Min-Hsiu; Chan, Wen-Ching; Shin, Jyh-Wei; Huang, Fu-Chin

    2016-07-01

    Acanthamoeba keratitis (AK) is a serious ocular disease caused by pathogenic Acanthamoeba gaining entry through wounds in the corneal injury; generally, patients at risk for contracting AK wear contact lenses, usually over a long period of time. Moreover, pathogenic Acanthamoeba causes serious consequences: it makes the cornea turbid and difficult to operate on, including procedures such as enucleation of the eyeball. At present, diagnosis of this disease is not straightforward, and treatment is very demanding. We have established the comparative transcriptome and extracellular secreted proteomic database according to the non-pathogenic strain ATCC 30010 and the pathogenic strains NCKU_B and NCKU_D. We identified 44 secreted proteins successfully, 10 consensus secreted proteins and 34 strain-specific secreted proteins. These proteins may provide targets for therapy and immuno-diagnosis of Acanthamoeba infections. This study shows a suitable approach to identify secreted proteins in Acanthamoeba and provides new perspectives for the study of molecules potentially involved in the AK.

  2. The effect of the disulfideisomerase domain containing protein in the defense against polyhexamethylene biguanide of highly tolerant Acanthamoeba at the trophozoite stage

    Directory of Open Access Journals (Sweden)

    Fu-Chin Huang

    2016-12-01

    Full Text Available Acanthamoeba castellanii is a free-living protozoan pathogen capable of causing a blinding keratitis and fatal granulomatous encephalitis. Current treatment generally involves an hourly application of polyhexamethylene biguanide (PHMB over a period of several days but this is not entirely effective against all strains/isolates. The tolerance mechanisms of PHMB in Acanthamoeba cells remain unclear. In this study, we found that the mRNA expression level of disulfideisomerase domain containing protein (PDI increased rapidly in surviving cells of the highly PHMB-tolerant Acanthamoeba castellanii strain, NCKH_D, during PHMB treatment, but not in the ATCC standard strain. After PDI-specific silencing, NCKH_D was found to be more vulnerable to PHMB treatment. The results described above show that PDI is an important gene for PHMB tolerance ability in a highly PHMB-tolerant strain of Acanthamoeba and provide a new insight for more efficient medicine development for Acanthamoeba keratitis.

  3. Targeting cyst wall is an effective strategy in improving the efficacy of marketed contact lens disinfecting solutions against Acanthamoeba castellanii cysts.

    Science.gov (United States)

    Abjani, Farhat; Khan, Naveed Ahmed; Yousuf, Farzana Abubakar; Siddiqui, Ruqaiyyah

    2016-06-01

    Acanthamoeba cysts are highly resistant to contact lens disinfecting solutions. Acanthamoeba cyst wall is partially made of 1,4 β-glucan (i.e., cellulose) and other complex polysaccharides making it a hardy shell that protects the resident amoeba. Here, we hypothesize that targeting the cyst wall structure in addition to antiamoebic compound would improve the efficacy of marketed contact lens disinfecting solutions. Using chlorhexidine as an antiamoebic compound and cellulase enzyme to disrupt cyst wall structure, the findings revealed that combination of both agents abolished viability of Acanthamoeba castellanii cysts and trophozoites. When tested alone, none of the agents nor contact lens disinfecting solutions completely destroyed A. castellanii cysts and trophozoites. The absence of cyst wall-degrading enzymes in marketed contact lens disinfecting solutions render them ineffective against Acanthamoeba cysts. It is concluded that the addition of cyst wall degrading molecules in contact lens disinfecting solutions will enhance their efficacy in decreasing the incidence of Acanthamoeba effectively.

  4. Induction of morphological and electrophysiological changes in hamster cornea after in vitro interaction with trophozoites of Acanthamoeba spp.

    Science.gov (United States)

    Omaña-Molina, Maritza; Navarro-García, Fernando; González-Robles, Arturo; Serrano-Luna, José de Jesús; Campos-Rodríguez, Rafael; Martínez-Palomo, Adolfo; Tsutsumi, Víctor; Shibayama, Mineko

    2004-06-01

    Acanthamoeba castellani and Acanthamoeba polyphaga are free-living amebae that cause keratitis and granulomatous encephalitis in humans. We have analyzed the early morphological and electrophysiological changes occurring during the in vitro interaction of cultured amebae with intact or physically damaged corneas obtained from hamsters. Both species of Acanthamoeba produced similar cytopathic changes, as seen by light microscopy and scanning electron microscopy. After adhesion to the epithelial surface, trophozoites formed clumps and migrated toward the cell borders, causing the separation of adjacent cells at 1 h of coculture. At later stages (2 to 4 h), some amebae were found under desquamating epithelial cells whereas others were seen associated with damaged cells or forming amebostome-like structures to ingest detached epithelial cells. Control corneas incubated in culture medium conditioned with amebae showed a cytoplasmic vacuolization and blurring of the epithelial-stromal junction. The early stages of corneal epithelial damage caused by amebae were also analyzed by measuring the transepithelial resistance changes in corneas mounted in Ussing chambers. Both species of Acanthamoeba caused a rapid decrease in electrical resistance. The present observations demonstrate that under in vitro conditions, Acanthamoeba trophozoites rapidly cause significant damage to the corneal epithelium. Furthermore, in our experimental model, previous physical damage to the corneas was not a prerequisite for the development of amebic corneal ulcerations.

  5. Quick survey for detection, identification and characterization of Acanthamoeba genotypes from some selected soil and water samples across Pakistan

    Directory of Open Access Journals (Sweden)

    Tania Tanveer

    2015-05-01

    Full Text Available [i]Acanthamoeba[/i] is an opportunistic protozoan pathogen which is widely distributed in nature and plays a pivotal role in ecosystem. [i]Acanthamoeba[/i] species may cause blinding keratitis and fatal granulomatous encephalitis involving central nervous system. In this study, we investigated the presence of [i]Acanthamoeba[/i] in soil and water resources of Pakistan. Here, [i]Acanthamoeba[/i] were recovered on non-nutrient agar plate lawn with [i]E.[/i] [i]coli[/i] and identified by morphological characteristics of the cyst. Furthermore PCR was performed with genus-specific primers followed by direct sequencing of the PCR product for molecular identification. Overall our PCR and sequencing results confirmed pathogenic genotypes including T4 and T15 from both soil and water samples. This is our first report of [i]Acanthamoeba[/i] isolation from both soil and water resources of Pakistan which may serve as a potential treat to human health across the country.

  6. High occurrence of Acanthamoeba genotype T4 in soil sources from Bolívar State, Venezuela.

    Science.gov (United States)

    Wagner, Carolina; Reyes-Batlle, María; Hernán, Aurora; Rojas, Elsy; Pérez, Gladymar; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

    2016-09-01

    Pathogenic strains of Acanthamoeba are causative agents of keratitis and encephalitis that often may end fatal in humans and other animals. In the present study, twenty-seven soil samples were collected in the Bolivar State in Venezuela and checked for the presence of Acanthamoeba. Samples were cultivated onto 2% non-nutrient agar plates seeded with a layer of heat killed E. coli. Amplification by PCR and sequencing of the DF3 region of the 18S rDNA of Acanthamoeba was carried out in order to confirm morphological identification of the amoebae. Furthermore, Acanthamoeba spp. was isolated from 51.8% of soil samples. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotype T4 in all samples. To the best of our knowledge, this is the first report of genotype T4 in soil sources from Venezuela. Further studies should be carried out in this State and in the country in order to determine the current occurrence of Acanthamoeba in Venezuelan environments.

  7. Synthesis, characterization and amoebicidal potential of locally synthesized TiO2 nanoparticles against pathogenic Acanthamoeba trophozoites in vitro.

    Science.gov (United States)

    Imran, Muhammad; Muazzam, Ambreen Gul; Habib, Amir; Matin, Abdul

    2016-06-01

    Acanthamoeba is an opportunistic protozoan pathogen that plays a pivotal role in the ecosystem. It may cause blinding keratitis and fatal encephalitis involving the central nervous system. Here we synthesized pure and Zn doped TiO2 nanoparticles (~10-30nm) via sol-gel and sol-hydrothermal methods and demonstrated its impact on the biological characteristics of pathogenic Acanthamoeba castellanii. Our results revealed that pure and Zn doped TiO2 nanoparticles synthesized by sol-hydrothermal methods (ranging 5, 10, 25 and 50μg/ml) exhibited amoebicidal effects i.e., >60% of trophozoites executed under normal light at maximum dose (50μg/ml) within 1h incubation. In contrast pure/doped TiO2 obtained via sol gel method showed ~40% amoeba damage. Furthermore, amoebae growth assay demonstrated that Zn doped TiO2 also inhibited Acanthamoeba numbers up to 7days in dose dependent manner. It was interesting to note that all the tested TiO2 nanoparticles have shown maximum amoebicidal effects at pH7 which is quite relevant to amoebic growth favorable conditions. Our results confirmed that TiO2 has inhibitory effects on Acanthamoeba growth and viability. Overall, we reported the amoebicidal and amoebic growth inhibition potential of pure and Zn doped TiO2 nanoparticles against Acanthamoeba due to attached OH(-) groups, reduced size and decreased band gap of sol hydrothermally synthesized TiO2 nanoparticles.

  8. Characterization of a new pathogenic Acanthamoeba Species, A. byersi n. sp., isolated from a human with fatal amoebic encephalitis.

    Science.gov (United States)

    Qvarnstrom, Yvonne; Nerad, Thomas A; Visvesvara, Govinda S

    2013-01-01

    Acanthamoeba spp. are free-living amoebae that are ubiquitous in natural environments. They can cause cutaneous, nasopharyngeal, and disseminated infection, leading to granulomatous amebic encephalitis (GAE) in immunocompromised individuals. In addition, they can cause amoebic keratitis in contact lens wearers. Acanthamoeba GAE is almost always fatal because of difficulty and delay in diagnosis and lack of optimal antimicrobial therapy. Here, we report the description of an unusual strain isolated from skin and brain of a GAE patient. The amoebae displayed large trophozoites and star-shaped cysts, characteristics for acanthamoebas belonging to morphology Group 1. However, its unique morphology and growth characteristics differentiated this new strain from other Group 1 species. DNA sequence analysis, secondary structure prediction, and phylogenetic analysis of the 18S rRNA gene confirmed that this new strain belonged to Group 1, but that it was distinct from the other sequence types within that group. Thus, we hereby propose the establishment of a new species, Acanthamoeba byersi n. sp. as well as a new sequence type, T18, for this new strain. To our knowledge, this is the first report of a Group 1 Acanthamoeba that is indisputably pathogenic in humans.

  9. Producción de citoquinas proinflamatorias durante los estadios tempranos de la infección experimental por Paracoccidioides brasiliensis

    OpenAIRE

    Luz Elena Cano; Angela Restrepo; Blanca Ortiz; Jorge Sahaza; Angel González

    2001-01-01

    La paracoccidioidomicosis (PCM) es una micosis sistémica restringida a América Latina y producida por la inhalación de las conidias del hongo dimórfico Paracoccidioides brasilienis (Pb) (1). En la PCM pulmonar experimental el hongo interactúa inicialmente con los macrófagos pulmonares residentes activándolos, lo que resulta en el
    reclutamiento de células inflamatorias como polimorfonucleares
    neutrófilos (PMN) (2). A su vez, las citoquinas proinflamatorias juegan

  10. Esclerosis múltiple en una mujer fallecida por quemaduras en el incendio del domicilio Multiple sclerosis in a woman died by burns in a house fire

    OpenAIRE

    J. Lucena; M. Salguero; A. Rico; Blanco, M.; Marín, R.; E. Barrero; F. Heredia

    2004-01-01

    La esclerosis múltiple (EM) o esclerosis en placas es la más común de las enfermedades inflamatorias desmielinizantes del SNC y el trastorno neurológico más incapacitante en los adultos jóvenes. Se caracteriza por la aparición de lesiones focales diseminadas en la sustancia blanca denominadas placas en las que lo más llamativo es la pérdida de la mielina (desmielinización) con preservación relativa de los axones. Es infrecuente que esta enfermedad entre en el campo de investigación médico-for...

  11. por valores

    Directory of Open Access Journals (Sweden)

    Jazmín Díaz-Barrios

    2005-01-01

    Full Text Available La era actual se caracteriza por una búsqueda de la identidad del hombre, donde el pivote del cambio organizacional es el individuo; cambiar en este entorno depende, en gran medida, de los valores que imperen en la cultura de la organización. Este trabajo documental intenta identificar aquellos valores, condición sine-qua-non, para el logro de los objetivos de cambio y analizarlos a fin de determinar sus indicadores, con el objeto de presentar una herramienta a las organizaciones que les facilite adaptarse a los nuevos tiempos. Se encontró que los valores: Delegación, Comunicación, Colaboración, Participación y Aprendizaje, son esenciales en los cambios integrales de esta era. Se determinaron 20 indicadores de su presencia. Se concluye que con esta información cada ente puede elaborar instrumentos que le permitan saber si los valores requeridos están presentes y en caso contrario tomar decisiones que alineen a la organización alrededor de los valores humanistas planteados, incrementando así las posibilidades de éxito en el proceso de cambio y por ende, las posibilidades de supervivencia en esta nueva época.

  12. Inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and application of statins as a novel effective therapeutic approach against Acanthamoeba infections.

    Science.gov (United States)

    Martín-Navarro, Carmen María; Lorenzo-Morales, Jacob; Machin, Rubén P; López-Arencibia, Atteneri; García-Castellano, José Manuel; de Fuentes, Isabel; Loftus, Brendan; Maciver, Sutherland K; Valladares, Basilio; Piñero, José E

    2013-01-01

    Acanthamoeba is an opportunistic pathogen in humans, whose infections most commonly manifest as Acanthamoeba keratitis or, more rarely, granulomatous amoebic encephalitis. Although there are many therapeutic options for the treatment of Acanthamoeba, they are generally lengthy and/or have limited efficacy. Therefore, there is a requirement for the identification, validation, and development of novel therapeutic targets against these pathogens. Recently, RNA interference (RNAi) has been widely used for these validation purposes and has proven to be a powerful tool for Acanthamoeba therapeutics. Ergosterol is one of the major sterols in the membrane of Acanthamoeba. 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase is an enzyme that catalyzes the conversion of HMG-CoA to mevalonate, one of the precursors for the production of cholesterol in humans and ergosterol in plants, fungi, and protozoa. Statins are compounds which inhibit this enzyme and so are promising as chemotherapeutics. In order to validate whether this enzyme could be an interesting therapeutic target in Acanthamoeba, small interfering RNAs (siRNAs) against HMG-CoA were developed and used to evaluate the effects induced by the inhibition of Acanthamoeba HMG-CoA. It was found that HMG-CoA is a potential drug target in these pathogenic free-living amoebae, and various statins were evaluated in vitro against three clinical strains of Acanthamoeba by using a colorimetric assay, showing important activities against the tested strains. We conclude that the targeting of HMG-CoA and Acanthamoeba treatment using statins is a novel powerful treatment option against Acanthamoeba species in human disease.

  13. Nutrición artificial en la insuficiencia intestinal: síndrome de intestino corto: Enfermedad inflamatoria intestinal Artificial nutrition in intestinal failure: short bowel syndrome. Intestinal inflammatory disease

    Directory of Open Access Journals (Sweden)

    T. Grau Carmona

    2005-06-01

    Full Text Available La resección intestinal amplia produce suficientes alteraciones como para requerir soporte nutricional especializado. Las medidas básicas de tratamiento, especial-mente en la fase aguda tras la resección intestinal o en presencia de complicaciones graves sobre pacientes con intestino corto, incluyen la repleción de fluidos y electro-litos y la instauración de soporte nutricional con el fin de prevenir la malnutrición. La nutrición enteral es el principal factor estimulador de la adaptación del intestino remanente. No obstante, su aplicación presenta dificultades en las fases agudas, por lo que los pacientes deben ser tratados frecuentemente con nutrición parenteral. La presencia de desnutrición puede ser también de importancia en los pacientes con enfermedad inflamatoria intestinal. El soporte nutricional está indicado en estos casos como tratamiento primario de la enfermedad, como tratamiento de la desnutrición o como tratamiento perioperatorio en los pacientes que requieren cirugía. A pesar de la patología digestiva, existen datos para recomendar la nutrición enteral como método inicial para el aporte de nutrientes en los pacientes que lo precisen.Large intestinal resection produces a sufficient number of impairments as to require specialized nutritional support. Basic treatment measures, especially during the acute phase after intestinal resection or in the presence of severe complications in patients with short bowel syndrome, include fluid and electrolytes reposition and nutritional support implementation in order to prevent hyponutrition. Enteral nutrition is the main stimulating factor for adaptation of the remaining bowel. However, its application has some difficulties during the acute phase, and thus patients must be frequently treated with parenteral nutrition. The presence of hyponutrition may be also important in patients with intestinal inflammatory disease. Nutritional support is indicated in these cases as the

  14. 18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong.

    Science.gov (United States)

    Booton, G C; Kelly, D J; Chu, Y-W; Seal, D V; Houang, E; Lam, D S C; Byers, T J; Fuerst, P A

    2002-05-01

    We examined partial 18S ribosomal DNA (Rns) sequences of Acanthamoeba isolates cultured in a study of microbial keratitis in Hong Kong. Sequence differences were sufficient to distinguish closely related strains and were used to examine links between strains obtained from corneal scrape specimens, contact lenses, lens cases, lens case solutions, and home water-supply faucets of patients with Acanthamoeba. We also looked for evidence of mixed infections. Identification of Acanthamoeba Rns genotypes was based on sequences of approximately 113 bp within the genus-specific amplicon ASA.S1. This permitted genotype identification by using nonaxenic cultures. Of 13 specimens obtained from corneal scrapes, contact lenses, lens cases, or lens case solutions, 12 were Rns genotype T4 and the remaining one was Rns genotype T3. The sequences of corneal scrape specimens of two patients also were the same as those obtained from their contact lenses or lens case specimens. A possible triple-strain infection was indicated by three different T4 sequences in cultures from one patient's lenses. Although faucet water used by patients to clean their lenses is a possible source of infections, specimens isolated from the faucets at two Acanthamoeba keratitis patients' homes differed from their corneal scrape or lens specimens. The overall results demonstrate the potential of this Rns region for tracking Acanthamoeba keratitis strains in infections and for distinguishing single-strain and closely related multiple-strain infections even when other microorganisms might be present with the cultured specimens. They also confirm the predominance of Rns genotype T4 strains in Acanthamoeba keratitis infections.

  15. Random amplified polymorphic DNA profiles as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba

    Directory of Open Access Journals (Sweden)

    Alves J.M.P.

    2000-01-01

    Full Text Available The genus Acanthamoeba comprises free-living amebae identified as opportunistic pathogens of humans and other animal species. Morphological, biochemical and molecular approaches have shown wide genetic diversity within the genus. In an attempt to determine the genetic relatedness among isolates of Acanthamoeba we analyzed randomly amplified polymorphic DNA (RAPD profiles of 11 Brazilian isolates from cases of human keratitis and 8 American type culture collection (ATCC reference strains. We found that ATCC strains belonging to the same species present polymorphic RAPD profiles whereas strains of different species show very similar profiles. Although most Brazilian isolates could not be assigned with certainty to any of the reference species, they could be clustered according to pattern similarities. The results show that RAPD analysis is a useful tool for the rapid characterization of new isolates and the assessment of genetic relatedness of Acanthamoeba spp. A comparison between RAPD analyses and morphological characteristics of cyst stages is also discussed.

  16. The use of dimethyl sulfoxide in contact lens disinfectants is a potential preventative strategy against contracting Acanthamoeba keratitis.

    Science.gov (United States)

    Siddiqui, Ruqaiyyah; Aqeel, Yousuf; Khan, Naveed Ahmed

    2016-10-01

    Acanthamoeba castellanii is the causative agent of blinding keratitis. Though reported in non-contact lens wearers, it is most frequently associated with improper use of contact lens. For contact lens wearers, amoebae attachment to the lens is a critical first step, followed by amoebae binding to the corneal epithelial cells during extended lens wear. Acanthamoeba attachment to surfaces (biological or inert) and migration is an active process and occurs during the trophozoite stage. Thus retaining amoebae in the cyst stage (dormant form) offers an added preventative measure in impeding parasite traversal from the contact lens onto the cornea. Here, we showed that as low as 3% DMSO, abolished A. castellanii excystation. Based on the findings, it is proposed that DMSO should be included in the contact lens disinfectants as an added preventative strategy against contracting Acanthamoeba keratitis. Copyright © 2016 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

  17. Prevalence of pathogenic Acanthamoeba (Protozoa:Amoebidae) in the atmosphere of the city of San Luis Potosi, Mexico.

    Science.gov (United States)

    Rodriguez-Zaragoza, S; Magana-Becerra, A

    1997-01-01

    Several species of pathogenic Acanthamoeba cause infections to humans, but amoebic keratitis is more frequently found than any other due to the increasing number of contact lens wearers in the world. Cysts and trophozoites of these amebas are airborne and may pollute water from the air. We investigated the proportion of pathogenic Acanthamoeba from the atmosphere of the city of San Luis Potosi. Samples were taken by the impinger method, every month during one year. We isolated 23 strains of Acanthamoeba, 61% of them were non-pathogenic, 31% were non-pathogenic with invasive capacity and 8% were pathogenic to mice. Almost 40% of these strains represent danger of infections to humans. The isolations were more abundant during the dry season in the south (urban) and west (suburban) stations, which means that the sanitary conditions around stands may enhance the proportion of pathogenic strains in the surroundings.

  18. Prevalence of Acanthamoeba spp. and other free-living amoebae in household water, Ohio, USA--1990-1992.

    Science.gov (United States)

    Stockman, Lauren J; Wright, Carolyn J; Visvesvara, Govinda S; Fields, Barry S; Beach, Michael J

    2011-03-01

    Knowledge of the prevalence of free-living amoebae (FLA) in US household water can provide a focus for prevention of amoeba-associated illnesses. Household water samples from two Ohio counties, collected and examined for amoebae during 1990-1992, were used to describe the prevalence of Acanthamoeba and other FLA in a household setting. Amoebae were isolated and identified by morphologic features. A total of 2,454 samples from 467 households were examined. Amoebae were found in water samples of 371 (79%) households. Sites most likely to contain amoeba were shower heads (52%) and kitchen sprayers (50%). Species of Hartmannella, Acanthamoeba, or Vahlkampfia were most common. Detection was higher in biofilm swab samples than in water samples. Detection of FLA and Acanthamoeba, at 79% and 51%, respectively, exceed estimates that have been published in previous surveys of household sources. We believe FLA are commonplace inhabitants of household water in this sample as they are in the environment.

  19. Effect of free versus liposomal-complexed pentamidine isethionate on biological characteristics of Acanthamoeba castellanii in vitro.

    Science.gov (United States)

    Siddiqui, Ruqaiyyah; Syed, Abdul; Tomas, Salvador; Prieto-Garcia, Jose; Khan, Naveed Ahmed

    2009-03-01

    Acanthamoeba is an opportunistic protozoan pathogen that can cause blinding keratitis and a rare but fatal encephalitis involving the central nervous system with a very poor prognosis. This is due to limited availability of effective anti-acanthamoebic drugs. Here, we tested whether the use of liposomes can improve the potency of pentamidine isethionate, an anti-amoebic compound. The liposomes consisted of l-alpha-phosphatidylcholine and cholesterol or ergosterol in a molar ratio of 1 : 5. Pentamidine isethionate was incorporated to achieve a final drug to lipid ratio of 1 : 5. At a drug concentration of 10 microg ml(-1), the liposomal drug was >12 times more effective than the free drug at preventing Acanthamoeba binding to human cells and significantly more effective in reducing parasite-mediated human cell cytopathogenicity, compared with the drug alone. Both the free and liposomal drug blocked Acanthamoeba encystation.

  20. Pathogenic Acanthamoeba spp secrete a mannose-induced cytolytic protein that correlates with the ability to cause disease.

    Science.gov (United States)

    Hurt, Michael; Neelam, Sudha; Niederkorn, Jerry; Alizadeh, Hassan

    2003-11-01

    The pathogenesis of Acanthamoeba keratitis begins when Acanthamoeba trophozoites bind specifically to mannosylated glycoproteins upregulated on the surfaces of traumatized corneal epithelial cells. When Acanthamoeba castellanii trophozoites are grown in methyl-alpha-D-mannopyranoside, they are induced to secrete a novel 133-kDa protein that is cytolytic to corneal epithelial cells. Clinical isolates of Acanthamoeba spp., and not the soil isolates, were proficient at producing a mannose-induced protein (MIP-133) and generating disease in Chinese hamsters. The purified protein was efficient at killing corneal epithelial cells, the first mechanistic barrier, by inducing apoptosis in a caspase 3-dependent pathway. Subsequent steps in pathogenesis require the amoebae to penetrate and degrade collagen. Only the clinical isolates tested were efficient at migrating through a collagenous matrix in vitro, presumably by MIP-133 degradation of both human type I and human type IV collagen. A chicken anti-MIP-133 antiserum effectively bound to the protein and blocked collagenolytic activity, migration, and cytopathic effects (CPE) against corneal cells in vitro. Chinese hamsters orally immunized with MIP-133 displayed a >30% reduction in disease. Immunoglobulin A isolated from immunized animals bound MIP-133 and blocked CPE on corneal cells in vitro. Animals induced to generate severe chronic infections displayed significant reductions in disease symptoms upon oral immunization postinfection. These data suggest that MIP-133 production might be necessary to initiate corneal disease and that it may play an important role in the subsequent steps of the pathogenic cascade of Acanthamoeba keratitis. Furthermore, as antibodies produced both prior to and after infection reduced clinical symptoms of disease, the protein may represent an important immunotherapeutic target for Acanthamoeba keratitis.

  1. Anti-Acanthamoeba Activities of Chloroformic Fractions of Trigonella Foenum Graecum (Seed and Their Cytotoxity on Mice Macrophage Cell

    Directory of Open Access Journals (Sweden)

    Samira Dodangeh

    2015-10-01

    Full Text Available Background: Acanthamoeba keratitis (AK is potentially a sight-threatening infection and its treatment is challenging. This is mainly due to presence of resistant cyst form. Indeed, cysts are highly resistant to current available drugs. Chemical drugs are toxic to human keratocytes. It should also be mentioned that most available anti-Acanthamoeba drugs are poorly cysticidal, In Iran and worldwide, AK cases continue to rise and therefore, novel effective drugs are urgently needed for the treatment of AK.Materials and Methods: In the present study, the in vitro activity of serial dilutions (10, 15, 20 and 25 mg/mL of chloroformic fractions including primary chloroformic fraction (minimum amount of chloroform, middle chloroformic fraction and remaining chloroformic fraction (most amount of chloroform of Trigonella foenum graecum seed were evaluated against Acanthamoeba trophozoites and cysts. Cytotoxic assay of fractions at different concentrations (25, 50, 100, 200, 300, 400, 500 mg/ml of test material was identified on mice Macrophage cells using MTT method.Results: The obtained results revealed that the tested fractions presented anti-amoebic activities in a time and dose dependent cycle. Anti-Acanthamoeba activity of remaining chloroformic fraction was more than other fractions. Trophozoites/cysts were eliminated when incubated with 15 and 20 mg/ml concentrations of remaining chloroformic fraction after 24 hours. Viability of macrophage cells was noted 100 % with 25 and 50 mg/ml concentration of remaining chloroformic fraction. Our results indicate that the plant fractions are safe for mammalian cells.Conclusion: Further studies should be performed in order to detect the active chemical compounds which could be used for the development of novel therapeutic approaches against Acanthamoeba infections. To the best of our knowledge, this is the first study on the activity of chloroformic fractions of Trigonella foenum graecum (seed against

  2. The role of domestic tap water on Acanthamoeba keratitis in non-contact lens wearers and validation of laboratory methods.

    Science.gov (United States)

    Koltas, Ismail Soner; Eroglu, Fadime; Erdem, Elif; Yagmur, Meltem; Tanır, Ferdi

    2015-09-01

    Acanthamoeba is increasingly recognized as an important cause of keratitis in non-contact lens wearers while contact lens wear is the leading risk factor for Acanthamoeba keratitis (AK). It is unlikely that the Acanthamoeba colonization is a feature which is effective only in patient's homes with infectious keratitis since the organism has been isolated from domestic tap water. Two hundred and thirty-one (231) corneal scrapings were taken from infectious keratitis cases, and four contact lens solutions and domestic tap waters were taken from 22 out of 44 AK-diagnosed patient's homes. Microscopic examination, culture, PCR, real-time PCR and DNA sequencing analyses were used for AK-diagnosed samples. The real-time PCR was the most sensitive (100 %) one among the methods used in diagnosis of AK. The 44 (19.0 %) out of 231 corneal scrapings, 4/4 (100 %) contact lens solution and 11/22 (50 %) of domestic tap water samples were found to be positive by real-time PCR for Acanthamoeba. A. griffini (T3), A. castellanii (T4) and A. jacobsi (T15) genotypes were obtained from corneal scrapings, contact lens solutions and domestic tap water samples taken from the patient's homes diagnosed with AK. The isolation of Acanthamoeba containing 6/22 (27.3 %) A. griffini (T3), 14/22 (63.6 %) A. castellanii (T4) and 2/22 (9.1 %) A. jacobsi (T15) from the domestic tap water outlets of 22 of 44 (50 %) of patient's homes revealed that is a significant source of these organisms. A. griffini (T3) and A. jacobsi (T15) genotypes have not been determined from AK cases in Turkey previously. Thus, we conclude that Acanthamoeba keratitis is associated with exposition of patients who has ocular trauma or ocular surface disease to domestic tap water in endemic or potentially endemic countries.

  3. Paralelismo clínico e imagenológico por resonancia magnética de los procesos inflamatorios de la médula espinal

    Directory of Open Access Journals (Sweden)

    Rómel Sarmiento Ducóngers

    2017-05-01

    Full Text Available Con el objetivo de identificar las características clínicas y por resonancia magnética de las lesiones inflamatorias de la médula espinal, se realizó un estudio observacional descriptivo transversal, de 56 pacientes, atendidos en los hospitales provinciales de Santiago de Cuba “Dr. Juan Bruno Zayas Alfonso” y “Dr. Saturnino Lora”, durante el periodo comprendido entre enero de 2014 hasta junio de 2015. Las características clínicas de las lesiones inflamatorias de la médula espinal fueron la cuarta década de la vida y el sexo masculino (60,7%, con síntomas predominantes como el dolor, la paresia y la parestesia. Existió predominio de las mielitis compresiva e inflamatoria. Al explorar el grado de concordancia entre la clínica y el informe de la resonancia magnética se obtuvo un índice de Kappa de 0,35. El diagnóstico imagenológico más relevante fue la mielitis compresiva en correspondencia con el diagnóstico clínico, no obstante la investigación permitió determinar que la concordancia entre la clínica y el diagnóstico imagenológico por resonancia magnética en los estudios realizados a estos pacientes en el periodo de estudio fue débil.

  4. Major role for cysteine proteases during the early phase of Acanthamoeba castellanii encystment.

    Science.gov (United States)

    Leitsch, David; Köhsler, Martina; Marchetti-Deschmann, Martina; Deutsch, Andrea; Allmaier, Günter; Duchêne, Michael; Walochnik, Julia

    2010-04-01

    Acanthamoeba castellanii is a facultative pathogen that has a two-stage life cycle comprising the vegetatively growing trophozoite stage and the dormant cyst stage. Cysts are formed when the cell encounters unfavorable conditions, such as environmental stress or food deprivation. Due to their rigid double-layered wall, Acanthamoeba cysts are highly resistant to antiamoebic drugs. This is problematic as cysts can survive initially successful chemotherapeutic treatment and cause relapse of the disease. We studied the Acanthamoeba encystment process by using two-dimensional gel electrophoresis (2DE) and found that most changes in the protein content occur early in the process. Truncated actin isoforms were found to abound in the encysting cell, and the levels of translation elongation factor 2 (EF2) were sharply decreased, indicating that the rate of protein synthesis must be low at this stage. In the advanced stage of encystment, however, EF2 levels and the trophozoite proteome were partly restored. The protease inhibitors PMSF (phenylmethylsulfonyl fluoride) and E64d [(2S,3S)-trans-epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester] inhibited the onset of encystment, whereas the protein synthesis inhibitor cycloheximide was ineffective. Changes in the protein profile, similar to those of encysting cells, could be observed with trophozoite homogenates incubated at room temperature for several hours. Interestingly, these changes could be inhibited significantly by cysteine protease inhibitors but not by inhibitors against other proteases. Taken together, we conclude that the encystment process in A. castellanii is of a bipartite nature consisting of an initial phase of autolysis and protein degradation and an advanced stage of restoration accompanied by the expression of encystment-specific genes.

  5. Acanthamoeba spp. as a universal host for pathogenic microorganisms: One bridge from environment to host virulence.

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    Guimaraes, Allan J; Gomes, Kamilla Xavier; Cortines, Juliana Reis; Peralta, José Mauro; Peralta, Regina H Saramago

    2016-12-01

    Free-living amoebas (FLA) are ubiquitous environmental protists that have enormously contributed to the microbiological contamination of water sources. FLAs have displayed resistance to environmental adversities and germicides and have played important roles in the population control of microbial communities due to its predatory behavior and microbicidal activity. However, some organisms have developed resistance to the intracellular milieu of amoebas, as in the case of Acanthamoebas, which in turn, have been functioning as excellent reservoirs for amoeba-resistant microorganisms (ARMs), such as bacteria, viruses and fungi. Little is known about these relationships and interaction mechanisms, but it is speculated that the FLAs need a very broad repertoire or universal class of receptors to bind and recognize these diverse species of microorganisms. By harboring these organisms as a "Trojan Horse", the Achantamoeba has been working as an excellent vector for pathogens. Moreover, studies have demonstrated that the interaction of pathogens with Acanthamoeba results in environmental selective pressure responsible for induction and maintenance of virulence factors and increase in microbial pathogenicity. This phenomenon is correlated to the observation of higher gene number and DNA content of ARMs, when compared to their relatives which are adapted to other hosts, due to allopatric or sympatric gene transfer and acquisition, contradicting the overall genome reduction theory for intracellularly adapted pathogens. Thus, adaptation to FLAs indirectly provided a "learning" environment for pathogens to resist later to macrophages; besides the evolutionary distance, these phagocytes share similar predatory mechanisms, such as phagocytosis and phagolysossomal degradation. In this mini-review, we cover the most important aspects of Acanthamoeba biology and their interactions with endemically important human pathogens.

  6. Results of case-control studies support the association between contact lens use and Acanthamoeba keratitis

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    Pacella E

    2013-05-01

    Full Text Available Elena Pacella,1 Giuseppe La Torre,2 Maria De Giusti,2 Chiara Brillante,1 Anna Maria Lombardi,2 Gianpaolo Smaldone,1 Tommaso Lenzi,1 Fernanda Pacella11Department of Sense Organs, Faculty of Medicine and Dentistry, Sapienza University of Rome, Italy; 2Department of Public Health and Infectious Diseases, Faculty of Pharmacy and Medicine, Sapienza University of Rome, ItalyBackground: Acanthamoeba keratitis (AK is ever more frequently reported in industrialized countries. The loss of the corneal surface integrity consequent to secondary microtrauma produced by the use of contact lens (CL favors the penetration of the parasite into the corneal tissue.Objectives: A scientific review was performed to investigate the association of CL wear as an Acanthamoeba keratitis (AK risk factor.Methods: A computerized screening of 7834 Medline articles (4623 from PubMed; 3211 from Scopus used a strict selection criteria of case-control studies involving CL wear and/or trauma.Results: The search yielded five case-control studies published from 1995 to 2012. All studies included showed a statistically significant positive association between AK and CL use, with a combined odds ratio (OR of 10.21 (95%, confidence intervals [CI]; 3.57–27.64.Statistical analysis: All studies included showed a statistically significant positive association between AK and CL use, though with differing OR values.Conclusion: Though rare, AK should be held in higher consideration when ophthalmologists are faced with CL users exhibiting simplex-like lesions associated with circular stromal infiltrates and disproportionate ocular pain in respect to the objective clinical picture.Keywords: keratitis, contact lens, Acanthamoeba

  7. Acanthamoeba encystment: multifactorial effects of buffers, biocides, and demulcents present in contact lens care solutions

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    Kovacs CJ

    2015-10-01

    Full Text Available Christopher J Kovacs, Shawn C Lynch, Marjorie J Rah, Kimberly A Millard, Timothy W Morris Bausch & Lomb Incorporated, Rochester, NY, USA Purpose: To determine whether agents which are purportedly capable of inducing encystment of Acanthamoeba can recapitulate the signal when tested in differing formulations. Methods: In accordance with the International Standard ISO 19045, Acanthamoeba castellanii ATCC 50370 trophozoites were cultured in antibiotic-free axenic medium, treated with test solutions, and encystment rates plus viability were measured via bright field and fluorescent microscopy. Test solutions included phosphate-buffered saline (PBS, borate-buffered saline, biguanide- and hydrogen peroxide (H2O2-based biocides, propylene glycol (PG and povidone (POV ophthalmic demulcents, and one-step H2O2-based contact lens disinfection systems. Results: Only PBS solutions with 0.25 ppm polyaminopropyl biguanide (PAPB and increasing concentrations of PG and POV stimulated A. castellanii encystment in a dose-dependent manner, whereas PBS solutions containing 3% H2O2 and increasing concentrations of PG and POV did not stimulate encystment. Borate-buffered saline and PBS/citrate solutions containing PG also did not stimulate encystment. In addition, no encystment was observed after 24 hours, 7 days, or 14 days of exposures of trophozoites to one-step H2O2 contact lens disinfection products or related solutions. Conclusion: The lack of any encystment observed when trophozoites were treated with existing or new one-step H2O2 contact lens care products, as well as when trophozoites were exposed to various related test solutions, confirms that Acanthamoeba encystment is a complex process which depends upon simultaneous contributions of multiple factors including buffers, biocides, and demulcents. Keywords: propylene glycol, contact lens care system, hydrogen peroxide disinfecting solution

  8. 'Candidatus Odyssella thessalonicensis' gen. nov., sp. nov., an obligate intracellular parasite of Acanthamoeba species.

    Science.gov (United States)

    Birtles, R J; Rowbotham, T J; Michel, R; Pitcher, D G; Lascola, B; Alexiou-Daniel, S; Raoult, D

    2000-01-01

    An intracellular bacterium, strain L13, was observed infecting an environmental isolate of an Acanthamoeba species. The bacterium could not be recovered on axenic medium but was recovered and cultivated in vitro using cultures of Acanthamoeba polyphaga. The 16S rRNA gene sequence of L13 was found to be new, sharing less than 84% similarity with other sequences in the GenBank/EMBL database. L13 was found to be a member of the alpha-Proteobacteria, sharing an evolutionary line of descent with a group of uniquely obligate intracellular organisms comprised of Caedibacter and Holospora species and the NHP bacterium. Viable bacteria appeared to be highly motile within amoebae. Ultrastructural analysis of the bacterium demonstrated that it is rod-shaped and possesses a typical Gram-negative cell wall, but has no other outstanding features except small vesicle-like structures often associated with the outer surface of each bacterium. The host range of L13 was found to be limited to the genus Acanthamoeba. In A. polyphaga, L13 infection was slow to manifest when cultures were incubated below 30 degrees C, but at higher temperatures bacteria multiplied prolifically and induced host cell lysis. The protein profile of the bacterium purified from the amoebae was assessed by SDS-PAGE and its G+C content was estimated to be 41 mol%. Although these results support the proposal of L13 as a new species, its obligate intracellular nature prevented isolation of a definitive type strain. L13 is therefore proposed as 'Candidatus Odyssella thessalonicensis' gen. nov., sp. nov.

  9. Comparison of hydrogen peroxide contact lens disinfection systems and solutions against Acanthamoeba polyphaga.

    Science.gov (United States)

    Hughes, R; Kilvington, S

    2001-07-01

    Acanthamoeba is a free-living amoeba causing a potentially blinding infection of the cornea. Contact lens wearers are most at risk and account for some 95% of cases. Hydrogen peroxide is used for contact lens disinfection due to its broad antimicrobial activity. Lenses must be neutralized before use to avoid pronounced stinging and possible corneal damage. Neutralization is achieved by adding a catalyst during the disinfection process (one-step) or afterwards (two-step). Here, the activities of commercial peroxide systems and individual solutions against trophozoites and cysts of Acanthamoeba polyphaga were compared. All disinfection systems were active against trophozoites, giving a > or = 3-log (99.9%) kill within 1 h. Of the four one-step systems, only one showed some cysticidal activity, giving a 1.28 +/- 0.41-log reduction. Both two-step systems were cysticidal, giving a > or = 3-log kill at 4 h. All system peroxide solutions were cysticidal, giving a > or = 3-log kill by 4 to 6 h. Variation in the cysticidal rate was observed with two solutions that gave a 1.8- to 2.1-log kill at 4 h compared with 3.0 to 4.0 for the rest (P < 0.05). No cysticidal activity was found with the peroxigen sodium perborate or the contact lens protein remover subtilisin A. Two-step systems are cysticidal providing contact times of at least 4 h are employed. Variation in cyst killing occurs between peroxide solutions, possibly due to formulation differences. One-step systems are less effective against Acanthamoeba cysts due to rapid peroxide neutralization. The cysticidal activity of one-step systems could be improved if neutralization rates were retarded.

  10. Evaluation of inhibitory potential of some selective methanolic plants extracts on biological characteristics of Acanthamoeba castellanii using human corneal epithelial cells in vitro.

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    Shoaib, Hafiz Muhammad; Muazzam, Ambreen Gul; Mir, Asif; Jung, Suk-Yul; Matin, Abdul

    2013-03-01

    Acanthamoeba is an opportunistic protozoan pathogen and known to be one of the most ubiquitous organisms, play a vital role in ecosystem, and recognized to cause blinding keratitis and rare but fatal granulomatous encephalitis involving the central nervous system with a very poor prognosis. This is due to limited availability of effective anti-Acanthamoeba drugs. The objective of the present study was to determine the efficacy of methanolic plants crude extracts on the viability and biological properties of Acanthamoeba castellanii (T4 genotype) and its cytotoxic effects on human corneal epithelial cells (HCEC). Using HCEC, it was observed that Acanthamoeba exhibited binding (>90 %) and cytotoxicity (>80 %) to host cells. However, plant crude extracts remarkably inhibited more than 70 and 60 % of Acanthamoeba binding and cytotoxicity to HCEC, respectively. It was further established that crude extracts (ranging from 0.1 to 1.5 mg/ml) exhibited amoebicidal effects, i.e., >50 % of trophozoites were killed/reduced at maximum dose (1.5 mg/ml) within 1 h incubation. However, the residual subpopulation remained static over longer incubations. Furthermore, growth assay demonstrated crude extracts inhibited >50 % Acanthamoeba numbers up to 7 days. Our results confirmed that plant crude extracts has inhibitory effects on Acanthamoeba growth and viability. Overall, these findings revealed that tested plant extracts is inhibitory to Acanthamoeba properties associated with pathogenesis. To the best of our knowledge, our findings demonstrated for the first time that selected methanol plant crude extracts exhibits inhibitory effects on biological properties of Acanthamoeba without any toxic effects on HCEC cells in vitro.

  11. Detection of free living amoebae, Acanthamoeba and Naegleria, in swimming pools, Malaysia.

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    Init, I; Lau, Y L; Arin Fadzlun, A; Foead, A I; Neilson, R S; Nissapatorn, V

    2010-12-01

    This study reports the detection of Acanthamoeba and Naegleria species in 14 swimming pools around Petaling Jaya and Kuala Lumpur, Malaysia. Sampling was carried out at 4 sites (the platforms (P), wall (W), 1 meter from the wall (1) and middle (2)) of each swimming pool. These free living amoebae (FLA) were detected under light and inverted microscopes after being cultured on the surface of non-nutrient agar lawned with Escherichia coli. Acanthamoeba species were detected in higher number of culture plates from all sampling sites of all the swimming pools. While Naegleria, were detected in fewer culture plates at 3 sampling sites (absent at site P) of 8 swimming pools. This suggested that the thick double-walled cysts of Acanthamoeba were more resistant, thus remaining viable in the dry-hot areas of the platforms and in chlorinated water of the swimming pools whereas Naegleria cysts, that are fragile and susceptible to desiccation, preferred watery or moist areas for growth and proliferation. The prevalence of both FLA was highest at site W (76.2%), followed by site 1 (64.7%), lowest at site 2 (19.4%), and could be detected at all 3 sampling levels (top, middle and bottom) of these 3 sites. The surface of site W might act as a bio-film that accumulated all kinds of microbes providing sufficient requirement for the FLA to develop and undergo many rounds of life cycles as well as moving from top to bottom in order to graze food. Other factors such as human activities, the circulating system which was fixed at all swimming pools, blowing wind which might carry the cysts from surroundings and the swimming flagellate stage of Naegleria could also contribute to the distribution of the FLA at these sampling sites. Both FLA showed highest growth (80.4%) at room temperature (25-28 ºC) and lesser (70.0%) at 37 ºC which might be due to the overgrowth of other microbes (E. coli, fungi, algae, etc). While at 44 ºC, only Acanthamoeba species could survive thus showing that

  12. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii

    OpenAIRE

    Shanan, Salah; Bayoumi, Magdi; Saeed, Amir; Sandström, Gunnar; Abd, Hadi

    2016-01-01

    Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11,...

  13. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii

    OpenAIRE

    Shanan, Salah; Bayoumi, Magdi; Saeed, Amir; Sandström, Gunnar; Abd, Hadi

    2016-01-01

    Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11,...

  14. Acanthamoeba Species Keratitis in a Soft Contact Lens Wearer Molecularly Linked to Well Water

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    Samira Mubareka

    2006-01-01

    Full Text Available Acanthamoeba species keratitis has been associated with soft contact lens wear. In the present report, an epidemiological link was established between the patient's isolate and well water from the home using molecular methods. To the authors' knowledge, this is the first case in Canada where such a link has been established. Primary care practitioners and specialists, including ophthalmologists and infectious diseases specialists, must maintain a high degree of clinical suspicion in soft contact lens wearers with keratitis unresponsive to conventional topical and systemic treatment.

  15. Acanthamoeba polyphaga mimivirus and other giant viruses: an open field to outstanding discoveries.

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    Abrahão, Jônatas S; Dornas, Fábio P; Silva, Lorena C F; Almeida, Gabriel M; Boratto, Paulo V M; Colson, Phillipe; La Scola, Bernard; Kroon, Erna G

    2014-06-30

    In 2003, Acanthamoeba polyphaga mimivirus (APMV) was first described and began to impact researchers around the world, due to its structural and genetic complexity. This virus founded the family Mimiviridae. In recent years, several new giant viruses have been isolated from different environments and specimens. Giant virus research is in its initial phase and information that may arise in the coming years may change current conceptions of life, diversity and evolution. Thus, this review aims to condense the studies conducted so far about the features and peculiarities of APMV, from its discovery to its clinical relevance.

  16. Enhanced Killing of Acanthamoeba Cysts with a Plant Peroxidase-Hydrogen Peroxide-Halide Antimicrobial System

    OpenAIRE

    Hughes, Reanne; Andrew, Peter W.; Kilvington, Simon

    2003-01-01

    The activity of H2O2 against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H2O2, and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05). With 2% H2O2, enhancement was observed at all ti...

  17. Molecular cloning and expression of phosphoglycerate dehydrogenase and phosphoserine aminotransferase in the serine biosynthetic pathway from Acanthamoeba castellanii.

    Science.gov (United States)

    Deng, Yihong; Wu, Duo; Tachibana, Hiroshi; Cheng, Xunjia

    2015-04-01

    Free-living amoebae of the genus Acanthamoeba are widespread protozoans that can cause serious infectious diseases. This study characterised phosphoglycerate dehydrogenase (PGDH) and phosphoserine aminotransferase (PSAT) in the phosphorylated serine biosynthetic pathway of Acanthamoeba castellanii. The PGDH gene encodes a protein of 442 amino acids with a calculated molecular weight of 47.7 kDa and an isoelectric point (pI) of 7.64. Meanwhile, the PSAT gene encodes a protein of 394 amino acids with a calculated molecular weight of 43.8 kDa and a pI of 5.80. Confocal microscopy suggests that PGDH is mainly diffused in the cytoplasm, whereas PSAT is located in the inner part of the cell membrane. The messenger RNA (mRNA) expression levels of PGDH and PSAT vary depending on growth state under consecutive culture conditions. No significant changes in the mRNA expression levels of both PGDH and PSAT occur after the incubation of L-serine with Acanthamoeba. This result indicates that exogenous serine exerts no influence on the expression of these genes and that the so-called feedback inhibition of both PGDH and PSAT in Acanthamoeba differs from that in bacteria or other organisms. We propose that the enzymes in the phosphorylated serine biosynthetic pathway function in amoeba growth and proliferation.

  18. Acanthamoeba belonging to T3, T4, and T11: genotypes isolated from air-conditioning units in Santiago, Chile.

    Science.gov (United States)

    Astorga, Berbeli; Lorenzo-Morales, Jacob; Martín-Navarro, Carmen M; Alarcón, Verónica; Moreno, Johanna; González, Ana C; Navarrete, Elizabeth; Piñero, José E; Valladares, Basilio

    2011-01-01

    Free-living amoebae (FLA) of the genus Acanthamoeba are widely distributed in the environment, in the air, soil, and water, and have also been isolated from air-conditioning units. The objective of this work was to investigate the presence of this genus of FLA in the air-conditioning equipment at the Institute of Public Health of Chile in Santiago, Chile. Water and air samples were collected from air-conditioning systems and were checked for the presence of Acanthamoeba spp. Positive samples were further classified at the genotype level after sequencing the highly variable diagnostic fragment 3 (DF3) region of the 18S rRNA gene. This is the first report of the T3, T4, and T11 genotypes of Acanthamoeba in air-conditioning units from Chile. Overall, the widespread distribution of potentially pathogenic Acanthamoeba strains in the studied source demands more awareness within the public and health professionals in Chile as this pathogen is emerging as a risk for human health worldwide. © 2011 The Author(s) Journal of Eukaryotic Microbiology © 2011 International Society of Protistologists.

  19. Monitoring of in vitro dynamics of Acanthamoeba strains isolated from infected eyes as a useful tool in keratitis management.

    Science.gov (United States)

    Chomicz, Lidia; Padzik, Marcin; Szaflik, Jacek P; Nahorski, Wacław L; Kryczka, Tomasz; Szaflik, Jerzy

    2014-11-01

    Free-living amoebae of Acanthamoeba genus are ubiquitous in various parts of the world. Some species of these amoebozoans present a serious risk to human health as the causative agents of vision-threatening diseases, Acanthamoeba keratitis. Correct diagnosis requires both a clinical examination of the cornea and amoebic form identification in affected eyes. Despite advances in pharmacotherapy, the infection is difficult to diagnose and to threat. Population dynamics of five different Acanthamoeba strains cultured in vitro under bacteria-free condition in BSC medium, was monitored in terms of diagnostic and therapeutic management. The range of protozoan number in the exponential growth phase, the morpho-physiological status of amoeba forms and their ability to multiply were evaluated. Results of the studies revealed that early and continued monitoring of the strains maintained in an axenic culture showed correlation between the dynamics of cultivated amoebae and the course of the disease, differences in response to pharmacotherapy and the surgical management efficacy. Concluding, the in vitro monitoring of dynamics of Acanthamoeba strains isolated from infected corneas may be important not only for proper diagnosis but also as a useful tool in keratitis management and therapeutic prognosis.

  20. Phylogenetic analysis and the evolution of the 18S rRNA gene typing system of Acanthamoeba.

    Science.gov (United States)

    Fuerst, Paul A; Booton, Gregory C; Crary, Monica

    2015-01-01

    Species of Acanthamoeba were first described using morphological characters including cyst structure and cytology of nuclear division. More than 20 nominal species were proposed using these methods. Morphology, especially cyst shape and size, has proven to be plastic and dependent upon culture conditions. The DNA sequence of the nuclear small-subunit (18S) rRNA, the Rns gene, has become the most widely accepted method for rapid diagnosis and classification of Acanthamoeba. The Byers-Fuerst lab first proposed an Rns typing system in 1996. Subsequent refinements, with an increasing DNA database and analysis of diagnostic fragments within the gene, have become widely accepted by the Acanthamoeba research community. The development of the typing system, including its current state of implementation is illustrated by three cases: (i) the division between sequence types T13 and T16; (ii) the diversity within sequence supertype T2/T6, and (iii) verification of a new sequence type, designated T20. Molecular studies make clear the disconnection between phylogenetic relatedness and species names, as applied for the genus Acanthamoeba. Future reconciliation of genetic types with species names must become a priority, but the possible shortcomings of the use of a single gene when reconstructing the evolutionary history of the acanthamoebidae must also be resolved.

  1. Biological characterization of a clinical and an environmental isolate of Acanthamoeba polyphaga: analysis of relevant parameters to decode pathogenicity.

    Science.gov (United States)

    da Rocha-Azevedo, Bruno; Costa e Silva-Filho, Fernando

    2007-11-01

    Acanthamoeba spp. consists of free-living amoebae, widespread in nature, which occasionally can cause human infections including granulomatous amoebic encephalitis and amoebic keratitis. Acanthamoeba pathogenesis is not entirely known and correlations between pathogenic potential and taxonomy are complex issues. In order to decipher the definition of a pathogenic amoeba, the objective of this work was to decipher the definition of pathogenic amoeba by characterizing two isolates of Acanthamoeba polyphaga obtained from different origins (a keratitis patient and freshwater), looking for differences among them. The clinical isolate grew faster in Peptone-yeast extract-glucose (PYG) medium, transformed more rapidly from a trophozoite to cyst and exhibited increased cytopathic effect on cultured cells. Morphological differences were also noted, since freshwater amoebae presented more acanthopodia than the clinical isolate. Moreover, actin labeling demonstrated that microfilament organization varies between isolates, with the presence of locomotory structures as lobopodia and lamellipodia in the keratitis isolate, which were less adherent on plastic. Zymography demonstrated that the keratitis isolates presented higher proteolytic activity and also were more able to invade collagen matrices. Altogether, we conclude that a group of stable physiological characteristics exist in Acanthamoeba that can be related to pathogenicity.

  2. Acanthamoeba feature a unique backpacking strategy to trap and feed on Listeria monocytogenes and other motile bacteria.

    Science.gov (United States)

    Doyscher, Dominik; Fieseler, Lars; Dons, Lone; Loessner, Martin J; Schuppler, Markus

    2013-02-01

    Despite its prominent role as an intracellular human pathogen, Listeria monocytogenes normally features a saprophytic lifestyle, and shares many environmental habitats with predatory protozoa. Earlier studies claimed that Acanthamoeba may act as environmental reservoirs for L. monocytogenes, whereas others failed to confirm this hypothesis. Our findings support the latter and provide clear evidence that L. monocytogenes is unable to persist in Acanthamoeba castellanii and A. polyphaga. Instead, external Listeria cells are rapidly immobilized on the surface of Acanthamoeba trophozoites, forming large aggregates of densely packed bacteria that we termed backpacks. While the assembly of backpacks is dependent on bacterial motility, flagellation alone is not sufficient. Electron micrographs showed that the aggregates are held together by filaments of likely amoebal origin. Time-lapse microscopy revealed that shortly after the bacteria are collected, the amoeba can change direction of movement, phagocytose the backpack and continue to repeat the process. The phenomenon was also observed with avirulent L. monocytogenes mutants, non-pathogenic Listeria, and other motile bacteria, indicating that formation of backpacks is not specific for L. monocytogenes, and independent of bacterial pathogenicity or virulence. Hence, backpacking appears to represent a unique and highly effective strategy of Acanthamoeba to trap and feed on motile bacteria.

  3. Response of first attack of inflammatory bowel disease requiring hospital admission to steroid therapy Respuesta al tratamiento esteroideo del primer brote de enfermedad inflamatoria intestinal que requiere ingreso hospitalario

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    M. Abu-Suboh Abadía

    2004-08-01

    Full Text Available Introduction: corticoid administration is the usual treatment of Crohn' disease (CD and ulcerative colitis (UC attacks. How-ever, information available on response rates and their predictive factors is scarce. Objective: to establish response to steroidal treatment in an homogeneous group of patients with CD or UC during their first admission to hospital. Methods: restrospective analysis of 86 patients who received systemic steroidal treatment for a severe flare-up during their first hospital admission between 1995 and 2000. Patients were treated per protocol with fluid therapy, absolute diet, IV 6-methyl-prednisolone 1 mg/kg/day, and enoxaparin at prophylactic doses. Clinical response at 30 days was considered good in case of complete remission, and poor in case of partial or absent remission. Univariate and multivariate analyses according to non-parametric statistics were performed for sociodemographic and biologic variables. Results: 45 patients with CD and 41 with UC were included. Good response rates were 64.4% for CD and 60.9% for UC. The univariate analysis showed that patients with good response have shorter evolution times and fewer previous flare-ups (p Introducción: la administración de corticoides constituye el tratamiento habitual de los brotes de enfermedad de Crohn (EC y colitis ulcerosa (CU. Sin embargo, existe poca información sobre las tasas de respuesta y los factores que puedan predecirla. Objetivo: determinar la respuesta al tratamiento esteroideo en un grupo homogéneo de pacientes de EC y CU que ingresan por primera vez en el hospital. Métodos: análisis retrospectivo de 86 pacientes que en su primer ingreso hospitalario recibieron tratamiento esteroideo sistémico por brote grave de EC o CU entre 1995 y 2000. Fueron tratados de forma protocolizada con sueroterapia, dieta absoluta, 6-metil-prednisolona 1 mg/kg/día IV y enoxaparina a dosis profilácticas. Se ha considerado la respuesta clínica a los 30 días como

  4. Evaluation of loop-mediated isothermal amplification assay for rapid diagnosis of Acanthamoeba keratitis

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    Abhishek Mewara

    2017-01-01

    Full Text Available Background: The clinical features of Acanthamoeba keratitis (AK are non-specific and closely resemble bacterial, viral and fungal keratitis. Materials and Methods: We compared loop-mediated isothermal amplification (LAMP with microscopy, non-nutrient agar (NNA culture and polymerase chain reaction (PCR in clinical suspects of AK. Results: Of 52 clinical samples (42 AK suspects and 10 proven bacterial, viral or fungal keratitis, 3 were positive by direct microscopy (sensitivity 60%, confidence interval [CI]: 17%–92.7%, and 5 by NNA culture, 18S rDNA PCR and LAMP (sensitivity 100%, CI: 46.3%–100%. The limit of detection of Acanthamoeba DNA was 1 pg/μl by both LAMP and PCR. Conclusion: PCR and LAMP assays targeting 18S rDNA gene were found particularly suitable for a rapid and accurate diagnosis of AK. LAMP assay takes 2–3 h lesser than PCR, and thus offers a rapid, highly sensitive and specific, simple and affordable diagnostic modality for patients suspected of AK, especially in resource limited settings

  5. Purification from Acanthamoeba castellanii of proteins that induce gelation and syneresis of F-actin.

    Science.gov (United States)

    Maruta, H; Korn, E D

    1977-01-10

    From Acanthamoeba castellanii, we have purified four proteins each of which alone causes a solution of F-actin to gel. The four active proteins have subunit molecular weights of about 23,000, 28,000, 32,000 and 38,000, respectively; the last three may be dimers in their native proteins. Together, these four proteins account for about 97% of the gelation activity of the whole extract; not more than about 3% of the total activity of the unfractionated extract can be due to a 250,000-dalton polypeptide. Another protein fraction, purified by agarose chromatography, induces shrinking (syneresis) of gels formed from F-actin and any of the gelation factors. That fraction contains a high Ca2+-, low (K+,EDTA)-ATPase and a major polypeptide of 170,000 daltons both of which bind to actin in the shrunken gel pellet. The active fraction does not contain the previously described Acanthamoeba myosin (Pollard, T. D., and Korn, E. D. (1973) J. Biol. Chem. 248, 4682-4690).

  6. Restriction fragment length polymorphisms of the DNA of selected Naegleria and Acanthamoeba amebae.

    Science.gov (United States)

    McLaughlin, G L; Brandt, F H; Visvesvara, G S

    1988-09-01

    Fourteen strains of Naegleria fowleri, two strains of N. gruberi, and one strain each of N. australiensis, N. jadini, N. lovaniensis, Acanthamoeba sp., A. castellanii, A. polyphaga, and A. comandoni isolated from patients, soil, or water were characterized by restriction fragment length polymorphisms. Total cellular DNA (1 microgram) was digested with either HindIII, BglII, or EcoRI; separated on agarose gels; and stained with ethidium bromide. From 2 to 15 unusually prominent repetitive restriction fragment bands, totaling 15 to 50 kilobases in length and constituting probably more than 30% of the total DNA, were detected for all ameba strains. Each species displayed a characteristic pattern of repetitive restriction fragments. Digests of the four Acanthamoeba spp. displayed fewer, less intensely staining repetitive fragments than those of the Naegleria spp. All N. fowleri strains, whether isolated from the cerebrospinal fluid of patients from different parts of the world or from hot springs, had repetitive restriction fragment bands of similar total lengths (ca. 45 kilobases), and most repetitive bands displayed identical mobilities. However, polymorphic bands were useful in identifying particular isolates. Restriction fragment length polymorphism analysis generally was consistent with taxonomy based on studies of infectivity, morphology, isoenzyme patterns, and antibody reactivity and suggests that this technique may help classify amebae isolated from clinical specimens or from the environment.

  7. Behavior of Yersinia enterocolitica in the presence of the bacterivorous Acanthamoeba castellanii.

    Science.gov (United States)

    Lambrecht, E; Baré, J; Van Damme, I; Bert, W; Sabbe, K; Houf, K

    2013-10-01

    Free-living protozoa play an important role in the ecology and epidemiology of human-pathogenic bacteria. In the present study, the interaction between Yersinia enterocolitica, an important food-borne pathogen, and the free-living amoeba Acanthamoeba castellanii was studied. Several cocultivation assays were set up to assess the resistance of Y. enterocolitica to A. castellanii predation and the impact of environmental factors and bacterial strain-specific characteristics. Results showed that all Y. enterocolitica strains persist in association with A. castellanii for at least 14 days, and associations with A. castellanii enhanced survival of Yersinia under nutrient-rich conditions at 25°C and under nutrient-poor conditions at 37°C. Amoebae cultivated in the supernatant of one Yersinia strain showed temperature- and time-dependent permeabilization. Intraprotozoan survival of Y. enterocolitica depended on nutrient availability and temperature, with up to 2.8 log CFU/ml bacteria displaying intracellular survival at 7°C for at least 4 days in nutrient-rich medium. Transmission electron microscopy was performed to locate the Yersinia cells inside the amoebae. As Yersinia and Acanthamoeba share similar ecological niches, this interaction identifies a role of free-living protozoa in the ecology and epidemiology of Y. enterocolitica.

  8. Silencing of xylose isomerase and cellulose synthase by siRNA inhibits encystation in Acanthamoeba castellanii.

    Science.gov (United States)

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2013-03-01

    A key challenge in the successful treatment of Acanthamoeba infections is its ability to transform into a dormant cyst form that is resistant to physiological conditions and pharmacological therapies, resulting in recurrent infections. The carbohydrate linkage analysis of cyst walls of Acanthamoeba castellanii showed variously linked sugar residues, including xylofuranose/xylopyranose, glucopyranose, mannopyranose, and galactopyranose. Here, it is shown that exogenous xylose significantly reduced A. castellanii differentiation in encystation assays (P < 0.05 using paired t test, one-tailed distribution). Using small interfering RNA (siRNA) probes against xylose isomerase and cellulose synthase, as well as specific inhibitors, the findings revealed that xylose isomerase and cellulose synthase activities are crucial in the differentiation of A. castellanii. Inhibition of both enzymes using siRNA against xylose isomerase and cellulose synthase but not scrambled siRNA attenuated A. castellanii metamorphosis, as demonstrated by the arrest of encystation of A. castellanii. Neither inhibitor nor siRNA probes had any effect on the viability and extracellular proteolytic activities of A. castellanii.

  9. Amino Acid Uptake and Metabolism of Legionella pneumophila Hosted by Acanthamoeba castellanii.

    Science.gov (United States)

    Schunder, Eva; Gillmaier, Nadine; Kutzner, Erika; Eisenreich, Wolfgang; Herrmann, Vroni; Lautner, Monika; Heuner, Klaus

    2014-07-25

    Legionella pneumophila survives and replicates within a Legionella-containing vacuole (LCV) of amoebae and macrophages. Less is known about the carbon metabolism of the bacteria within the LCV. We have now analyzed the transfer and usage of amino acids from the natural host organism Acanthamoeba castellanii to Legionella pneumophila under in vivo (LCV) conditions. For this purpose, A. castellanii was 13C-labeled by incubation in buffer containing [U-(13)C(6)]glucose. Subsequently, these 13C-prelabeled amoebae were infected with L. pneumophila wild type or some mutants defective in putative key enzymes or regulators of carbon metabolism. 13C-Isotopologue compositions of amino acids from bacterial and amoebal proteins were then determined by mass spectrometry. In a comparative approach, the profiles documented the efficient uptake of Acanthamoeba amino acids into the LCV and further into L. pneumophila where they served as precursors for bacterial protein biosynthesis. More specifically, A. castellanii synthesized from exogenous [U-13C6]glucose unique isotopologue mixtures of several amino acids including Phe and Tyr, which were also observed in the same amino acids from LCV-grown L. pneumophila. Minor but significant differences were only detected in the isotopologue profiles of Ala, Asp, and Glu from the amoebal or bacterial protein fractions, respectively, indicating partial de novo synthesis of these amino acids by L. pneumophila. The similar isotopologue patterns in amino acids from L. pneumophila wild type and the mutants under study reflected the robustness of amino acid usage in the LCV of A. castellannii.

  10. Functional expression and characterization of an iron-containing superoxide dismutase of Acanthamoeba castellanii.

    Science.gov (United States)

    Kim, Jung-Yeon; Na, Byoung-Kuk; Song, Kyoung-Ju; Park, Mi-Hyun; Park, Yun-Kyu; Kim, Tong-Soo

    2012-10-01

    Acanthamoeba spp. are free-living amoebae, but opportunistic infections of some strains of the organisms cause severe diseases such as acanthamoebic keratitis, pneumonitis, and granulomatous amoebic encephalitis in human. In this study, we identified a gene encoding iron superoxide dismutase of Acanthamoeba castellanii (AcFe-SOD) and characterized biochemical and functional properties of the recombinant enzyme. Multiple sequence alignment of the deduced amino acid sequence of AcFe-SOD with those of previously reported iron-containing SODs (Fe-SODs) from other protozoan parasites showed that AcFe-SOD shared common metal-binding residues and motifs that are conserved in Fe-SODs. The genomic length of the AcFe-SOD gene was 926 bp consisting of five exons interrupted by four introns. The recombinant AcFe-SOD showed similar biochemical characteristics with its native enzyme and shared typical biochemical properties with other characterized Fe-SODs, including molecular structure, broad pH optimum, and sensitivity to hydrogen peroxide. Immunolocalization analysis revealed that the enzyme localized in the cytosol of the trophozoites. Activity and expression level of the enzyme were significantly increased under oxidative stressed conditions. These results collectively suggest that AcFe-SOD may play essential roles in the survival of the parasite not only by protecting itself from endogenous oxidative stress but also by detoxifying oxidative killing of the parasite by host immune effector cells.

  11. Temperature limitation may explain the containment of the trophozoites in the cornea during Acanthamoeba castellanii keratitis.

    Science.gov (United States)

    Nielsen, Mattias Kiel; Nielsen, Kim; Hjortdal, Jesper; Sørensen, Uffe B Skov

    2014-12-01

    Acanthamoeba keratitis is a serious sight-threatening disease. The relatively low temperature of the cornea may explain why amoebic infections usually are localized in this tissue and rarely spread to other parts of the eye. In this study, the growth rate of the amoeba Acanthamoeba castellanii was examined at different temperatures. The aim was to establish the optimal growth temperature for A. castellanii and to examine the growth within the vicinity of the core body temperature. The growth rates of four clinical and two environmental strains of A. castellanii were estimated at different temperatures, and temperature limitations for the trophozoite stage was established. Movements influenced by temperature gradients were monitored for two clinical strains of A. castellanii. The highest growth rate for each of the six amoebic strains tested was found to be close to 32 °C. The growth of the trophozoites of all examined strains was greatly reduced or completely halted at temperatures above 36 °C and encysted at the elevated temperature. Thus, the optimal growth temperature for the four strains of A. castellanii is close to the surface temperature of the human cornea, while the higher body core-temperature induced encysting of the amoebae. This may explain why most amoebic eye infections are confined to the cornea.

  12. Potent antimicrobial peptides against Legionella pneumophila and its environmental host, Acanthamoeba castellanii.

    Science.gov (United States)

    Schlusselhuber, Margot; Humblot, Vincent; Casale, Sandra; Méthivier, Christophe; Verdon, Julien; Leippe, Matthias; Berjeaud, Jean-Marc

    2015-06-01

    Legionella pneumophila, the major causative agent of Legionnaires' disease, is most often found in the environment in close association with free-living amoebae, leading to persistence, spread, biocide resistance, and elevated virulence of the bacterium. In the present study, we evaluated the anti-Legionella and anti-Acanthamoeba activities of three alpha-helical antimicrobial peptides (AMPs), namely, NK-2, Ci-MAM-A24, and Ci-PAP-A22, already known for the extraordinary efficacy against other microbes. Our data represent the first demonstration of the activity of a particular AMP against both the human facultative intracellular pathogen L. pneumophila and its pathogenic host, Acanthamoeba castellanii. Interestingly, the most effective peptide, Ci-MAM-A24, was also found to reduce the Legionella cell number within amoebae. Accordingly, this peptide was immobilized on gold surfaces to assess its antimicrobial activity. Surfaces were characterized, and activity studies revealed that the potent bactericidal activity of the peptide was conserved after its immobilization. In the frame of elaborating anti-Legionella surfaces, Ci-MAM-A24 represents, by its direct and indirect activity against Legionella, a potent peptide template for biological control of the bacterium in plumbings.

  13. Mechanism of cyst specific protein 21 mRNA induction during Acanthamoeba differentiation.

    Science.gov (United States)

    Chen, Li; Orfeo, Tom; Gilmartin, Greg; Bateman, Erik

    2004-04-01

    The Acanthamoeba cyst specific protein 21 (CSP21) gene is tightly repressed in growing cells and highly induced early during differentiation into a dormant cyst. This increase is mediated by the rate of transcription of the CSP21 gene as determined by nuclear run-on assays. The promoter region of the CSP21 gene was analyzed by transcript start site mapping and in vitro transcription of wild-type or mutant templates, using extracts from growing cells. A sequence located 3' to a modified TATA box completely inhibits transcription and removal of this region permits robust transcription utilizing a start site approximately 35 base pairs downstream of the TATA box. Sequences 5' to the TATA box had no effect on transcription, suggesting that anti-repression is the only mechanism required for CSP21 induction. Fractionation of nuclear extracts yielded a fraction capable of transcription from the CSP21 promoter, and a fraction containing a promoter-specific repressing activity. Anti-repression may thus be a major mechanism regulating differentiation or maintenance of the proliferative cycle in Acanthamoeba.

  14. Acanthamoeba keratitis update-incidence, molecular epidemiology and new drugs for treatment.

    Science.gov (United States)

    Seal, D V

    2003-11-01

    A reliable figure for the expected incidence of Acanthamoeba keratitis of one per 30000 contact lens wearers per year has now been obtained from a combination of three cohort and three Questionnaire Reporting Surveys; 88% of cases wore hydrogel lenses and 12% wore rigid lenses. This figure now provides a basis for the expected number of cases against which to judge either epidemic outbreaks or effects of prevention with disinfecting solutions, better hygiene, or the use of disposable lenses. Molecular biology of Acanthamoeba has advanced considerably in the last 10 years with new automated sequencing technology. This has allowed the construction of a genotype identification scheme with 13 different genotypes against which to compare clinical isolates for epidemiological investigations or pathogenicity markers. So far, only four genotypes have been associated with keratitis of which the majority have been T4 but T3, T6, and T11 have each caused individual cases. Each genotype is heterogenous and can be further subdivided by comparison of sequences of diagnostic fragments of 18S rDNA, riboprinting by PCR-RFLP of 18S rDNA, or by mitochondrial DNA RFLP. Drug therapy has been revolutionised with the introduction of the biguanides-chlorhexidine or polyhexamethylene biguanide-with most but not all infections quickly resolving. Failure can still occur occasionally and further research is needed on more effective combination chemotherapy. A number of guanidines have been identified in this paper that could be usefully pursued as part of combination chemotherapy along with the alkylphosphocholines.

  15. Investigating the role of Acanthamoeba polyphaga in protecting Human Adenovirus from water disinfection treatment.

    Science.gov (United States)

    Verani, Marco; Di Giuseppe, Graziano; Tammaro, Carmine; Carducci, Annalaura

    2016-06-01

    Human adenoviruses are responsible for a wide range of clinical infections and are present in aquatic environments, including river, seawater, drinking-water and sewage. Free-living amoebae (Acanthamoeba) in the same environments may internalize them and other microorganisms can act as a reservoir for the internalized viruses. In this study, we studied the interaction between Acanthamoeba polyphaga and Human Adenovirus type 5 (HAdV 5) to determine whether the amoeba played a role in protecting the internalized viruses from chemical disinfection. The efficacy of sodium hypochlorite disinfection against A. polyphaga and HAdV 5 either singly or in combination was assessed at three different concentrations. Individually, the amoeba were more resistant to chemical disinfection than HAdV 5 and remained alive after exposure to 5mg/l of sodium hypochlorite. In contrast, HAdV 5 lost infectivity following exposure to 2.5mg/l of sodium hypochlorite. When the amoeba and HAdV 5 were co-cultured, infectious virus was found in the cytoplasm of the amoeba at 5mg/l disinfectant concentration. These findings suggest that the A. polyphaga is providing protection for the HAdV 5.

  16. Morphological Features and In Vitro Cytopathic Effect of Acanthamoeba griffini Trophozoites Isolated from a Clinical Case

    Directory of Open Access Journals (Sweden)

    Arturo González-Robles

    2014-01-01

    Full Text Available Light and transmission electron microscopy observations are reported on the structure and in vitro cytopathic effect of Acanthamoeba griffini trophozoites isolated from a clinical case. Live trophozoites were moderately active with a remarkable pleomorphism which changed from ovoid to quite elongated shapes. When moving, amoebae formed cytoplasmic projections such as wide lamellae and acanthopodia of diverse size and thickness which contain a significant amount of actin. Ultrastructurally, the cytoplasm showed the main organelles found in other free-living amoebae. Coincubation of trophozoites with MDCK cell monolayers resulted in a local damage to target cells after 24 h of interaction, suggesting that the cytopathic effect is contact-dependent. By transmission electron microscopy, amoebae appeared to engulf small portions of the MDCK cells; however, the cells that were not in contact with trophozoites had an unaltered morphology. When epithelial monolayers were incubated with conditioned medium for 24 h, small areas of cell injury were also observed. The phylogenetical analysis as well as the sequencing of the acquired amplified product for the DF3 region of the amoebae isolate confirmed that it belongs to genotype T3, which includes other pathogenic amoebae; besides the activity of two drugs currently used against Acanthamoeba was tested on A. griffini.

  17. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

    Directory of Open Access Journals (Sweden)

    Jenny Olofsson

    Full Text Available The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced.

  18. Proteinase activities in total extracts and in medium conditioned by Acanthamoeba polyphaga trophozoites.

    Science.gov (United States)

    Alfieri, S C; Correia, C E; Motegi, S A; Pral, E M

    2000-04-01

    Acanthamoeba species can cause granulomatous encephalitis and keratitis in man. The mechanisms that underlie tissue damage and invasion by the amoebae are poorly understood, but involvement of as yet uncharacterized proteinases has been suggested. Here, we employed gelatin-containing gels and azocasein assays to examine proteinase activities in cell lysates and in medium conditioned by Acanthamoeba polyphaga trophozoites. Azocasein hydrolysis by cell lysates was optimally detected at pH 4.0-5.0 and was predominantly associated with the activity of cysteine proteinases. Compatible with enzyme activation during secretion, culture supernatants additionally contained a prominent azocasein hydrolyzing activity attributable to serine proteinases; these enzymes were better detected at pH 6.0 and above, and resolved at 47, 60, 75, 100, and >110 kDa in overlay gelatin gels. Although a similar banding profile was observed in gels of trophozoite lysates, intracellular serine proteinases were shown to be activated during electrophoresis and to split the substrate during migration in sodium dodecyl sulfate gels. Blockage of serine proteinases with phenylmethylsulfonylfluoride prior to electrophoresis permitted the detection of 43-, 59-, 70-, and 100-130-kDa acidic cysteine proteinases in cell lysates, and of 3 (43, 70, and 130 kDa) apparently equivalent enzymes in culture supernatants. Under the conditions employed, no band associated with a metalloproteinase activity could be depicted in substrate gels, although the discrete inhibition of supernatants' azocaseinolytic activity by 1,10-phenanthroline suggested secretion of some metalloproteinase.

  19. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

    Science.gov (United States)

    Olofsson, Jenny; Axelsson-Olsson, Diana; Brudin, Lars; Olsen, Björn; Ellström, Patrik

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced.

  20. Influence of Legionella pneumophila and other water bacteria on the survival and growth of Acanthamoeba polyphaga.

    Science.gov (United States)

    Anacarso, I; Guerrieri, E; Bondi, M; de Niederhäusern, S; Iseppi, R; Sabia, C; Contri, M; Borella, P; Messi, P

    2010-10-01

    We investigated in solid medium, in water microcosm co-cultures and by light and transmission electron microscopy the influence of Legionella pneumophila Lp-1, Pseudomonas aeruginosa ATCC 27853, Burkholderia cepacia ATCC 25416 and Pseudomonas fluorescens SSD35 on the growth and survival of Acanthamoeba polyphaga. The infection with L. pneumophila was microscopically characterized by the presence of few bacteria inside protozoa at 4th h, and by the beginning of disruptive effects in late phase of trial. In water microcosm studies, performed at different temperature, the more significant interactions were observed at 30°C. In these conditions, L. pneumophila caused a marked reduction in trophozoite and cyst counts from the 4th day until the end of incubation (11 days). B. cepacia showed, by microscopic observation, few and generally single rods within protozoan phagosomes and caused a light reduction of trophozoite viability and cyst formation in co-cultures. A more invasive type of endocytosis, characterized by an early invasion with the presence of a high bacteria number inside amoebae, was observed for Pseudomonas strains. P. fluorescens produced a violent lysis of the host, whereas P. aeruginosa did not cause lysis or suffering. These results underline that water bacteria other than legionella are capable of intracellular survival in Acanthamoeba, influencing the protozoa viable cycle.