Modified amoebic liver abscess: Case report / Absceso hepático amebiano modificado: reporte de un caso

Abstract We present the case of a 56-year-old black female patient from a rural area in the Morón municipality, Ciego de Ávila province, Cuba. She was admitted with symptoms of dysentery with several days of evolution and a later episode of high fever, compromised general status, and abdominal pain located in the right hypochondrium. Analytical studies reported leukocytosis with a predominance of polymorphonuclear cells, Entamoeba histolytica was found in the stool study. Abdominal ultrasound reported a mixed image of 110 x 84 mm in the upper right lobe of the liver, as confirmed by computed tomography. This image was interpreted as a possible liver abscess. The patient received antimicrobial treatment for four weeks without a good response, thus requiring surgical intervention. She evolved favorably and was discharged after 21 days.

Resumen Se presenta el caso de una paciente de raza negra de 56 años procedente de área rural de Morón, provincia Ciego de Ávila (Cuba), quien ingresa por cuadro clínico de disentería de varios días de evolución acompañado de fiebre, compromiso de su estado general y dolor abdominal en el hipocondrio derecho. Los estudios analíticos de laboratorio mostraron leucocitosis con predominio de neutrófilos y presencia de trofozoitos de Entamoeba histolytica en la materia fecal. La ecografía de abdomen reporto una imagen mixta de 110 x 84 mm en el lóbulo derecho del hígado y la tomografía confirmó la lesión que se interpretó como un posible absceso hepático. Se inició tratamiento antimicrobiano por un periodo de 4 semanas sin adecuada respuesta por lo que requirió tratamiento quirúrgico. Su evolución fue favorable con egreso a los 21 días.

Amebiasis intestinal y hepática / Intestinal and hepatic amebiasis

Amebiasis is the infection by Entamoeba histolytica, a protozoan capable of invading the colonic mucosa causing a diarrheic syndrome, although most of the time is mild, it can lead to a fulminating colitis. Sometimes it can spread to other organs; among extra-intestinal manifestations of this parasite, the most frequent is the amebic liver abscess. In the next pages, general aspects of this protozoan, its epidemiology, clinical findings, diagnosis and treatment are reviewed, emphasizing the possibilities of diagnosis and treatment available in Chile.

La amebiasis corresponde a la infección por Entamoeba histolytica, protozoo capaz de invadir la mucosa del colon provocando un cuadro diarréico que, si bien la mayoría de las veces es leve, puede llegar a una colitis fulminante. En algunas oportunidades puede diseminarse a otros órganos; dentro de las manifestaciones extra-intestinales de este parásito, la más frecuente es el absceso hepático amebiano. A continuación se revisan aspectos generales de este protozoo, su epidemiología, cuadro clínico, diagnóstico y tratamiento, destacando las posibilidades de diagnóstico y tratamiento disponibles en Chile.

Parasitosis y síndrome de intestino irritable / Parasitosis and irritable bowel syndrome

El síndrome de intestino irritable (SII) es un trastorno funcional digestivo de etiología multifactorial. En su fisiopatología se describen diversos factores, tanto biológicos, como psicológicos y ambientales, que afectan el estado de activación de células inmunes en la mucosa intestinal. Entre los factores ambientales se incluye la presencia de alguna parasitosis intestinal. El síndrome de intestino irritable post-infeccioso (SII-PI) es reconocido como un subgrupo de estos trastornos, cuya aparición de los síntomas es posterior a una infección intestinal provocada por agentes microbianos. A pesar de que en Chile hay pocos estudios respecto a la relación entre SII y parasitosis intestinal, se ha descrito la existencia de una asociación positiva entre SII e infecciones por Blastocistis hominis, uno de los parásitos prevalentes en Chile. En otros países, se ha descrito además una relación entre SII, amebiasis y giardiasis. Por la alta prevalencia de parasitosis en nuestro país, existe la necesidad de ampliar los estudios para clarificar la fuerza de la asociación entre parasitosis y SII.

Irritable bowel syndrome (IBS) is a functional disorder of the gastrointestinal tract characterised by multi-factorial aetiology. In IBS physiopathology are involved diverse factors between them biological, psychosocial, and environmental components which affect the immune activation status of gut mucosa. Among these factors is recognized the intestinal parasitosis. Post-infection IBS (PI-IBS) is recognised as a subgroup of functional disorders whose symptoms onset appear after a symptomatic intestinal infection caused by microbial agents. There are few studies regarding of relationship between IBS and intestinal parasitosis in Chile. However, is has been well described a positive association between IBS and Blastocystis hominis infections, one of prevalent parasites in Chile. In other countries, is also described a relationship between IBS and amebiasis and giardiasis. Both, characterized by a common mode of transmission through water as well as contaminated food. Because the high prevalence of parasitosis in our country it is necessary to expand the association studies to clarify the strength of the parasites ethiology in IBS.

Amebiasis: aspectos clínicos, terapéuticos y de diagnóstico de la infección / An update on amebiasis

The description of Entamoeba dispar, and the recovery of Entamoeba moshkovskii from humans had a major impact in the epidemiology and clinical management of amebiasis. Infections range from asymptomatic colonization to hemorrhagic colitis and extra-intestinal diseases. Only a minority of amebiasis patients progress to the development of disease. Recent studies suggest that susceptibility to infection, and its outcome is influenced by the host, parasite genotype, and environment. The identification of Entamoeba histolytica is based on the detection of specific antigens by ELISA and DNA in stool and other clinical samples. Several diagnostic tests have been developed, including polymerase chain reaction, the technique of choice, for the detection and differentiation of E. histolytica, E. dispar, and E. moshkovskii. Combination of serologic tests with detection of the parasite DNA by PCR or antigen by ELISA offers the best approach to diagnosis. However, these techniques are impractical for clinical laboratories of developing countries. Clinicians must follow the guidelines of the World Health Organization to avoid unnecessary treatments. This review describes and discusses recent advances in amebiasis with emphasis in the clinical aspects and management of infection.

Entamoeba histolytica Induces Cell Death of HT29 Colonic Epithelial Cells via NOX1-Derived ROS

Entamoeba histolytica, which causes amoebic colitis and occasionally liver abscess in humans, is able to induce host cell death. However, signaling mechanisms of colon cell death induced by E. histolytica are not fully elucidated. In this study, we investigated the signaling role of NOX in cell death of HT29 colonic epithelial cells induced by E. histolytica. Incubation of HT29 cells with amoebic trophozoites resulted in DNA fragmentation that is a hallmark of apoptotic cell death. In addition, E. histolytica generate intracellular reactive oxygen species (ROS) in a contact-dependent manner. Inhibition of intracellular ROS level with treatment with DPI, an inhibitor of NADPH oxidases (NOXs), decreased Entamoeba-induced ROS generation and cell death in HT29 cells. However, pan-caspase inhibitor did not affect E. histolytica-induced HT29 cell death. In HT29 cells, catalytic subunit NOX1 and regulatory subunit Rac1 for NOX1 activation were highly expressed. We next investigated whether NADPH oxidase 1 (NOX1)-derived ROS is closely associated with HT29 cell death induced by E. histolytica. Suppression of Rac1 by siRNA significantly inhibited Entamoeba-induced cell death. Moreover, knockdown of NOX1 by siRNA, effectively inhibited E. histolytica-triggered DNA fragmentation in HT29 cells. These results suggest that NOX1-derived ROS is required for apoptotic cell death in HT29 colon epithelial cells induced by E. histolytica.

Mecanismos específicos de patogenicidadede protozoários de mucosa: Entamoeba histolytica, Giardia lamblia e Trichomonas vaginali / Specific pathogenic mechanisms of mucosal protozoans: Entamoeba histolytica, Giardia lamblia and Trichomonas vaginalis

Entamoeba histolytica e Giardia lamblia são protozoários que podem parasitar a mucosa intestinal, causando principalmente diarreia. Trichomonas vaginalis coloniza a mucosa vaginal causando tricomonose, a doença sexualmente transmissível não viral mais comum no mundo. Embora coletivamente estes parasitos infectem mais de um bilhão de pessoas a cada ano, seus mecanismos de patogenicidade ainda não estão totalmente esclarecidos. Assim, esta revisão reúne os principais mecanismos envolvidos na patogenicidade destes protozoários, bem como os fatores do microambiente que podem interferir no sucesso da colonização. A patogênese da E. histolytica envolve adesão, lise, fagocitose de células epiteliais e bactérias, invasão tecidual por ação de enzimas e evasão da resposta imune do hospedeiro. A lectina Gal/GalNAc, os amebaporos e as cisteína proteases são as principais moléculas envolvidas nesses processos. O estabelecimento da giardiose depende de diversos mecanismos patogênicos e de virulência desenvolvidos pela G. lamblia, tais como as moléculas envolvidas na adesão, encistamento e variação antigênica. Para o sucesso da colonização da mucosa vaginal, o T. vaginalis expressa moléculas como as adesinas de superfície, lipofosfoglicanos e galectina, envolvidas na adesão às células epiteliais vaginais e alteração da expressão gênica, tanto do parasito como do hospedeiro.

Entamoeba histolytica and Giardia lamblia are protozoans that may parasitize the intestinal mucosa, mainly causing diarrhea. Trichomonas vaginalis colonizes the vaginal mucosa causing trichomonosis, the most common non-viral sexually transmitted disease in the world. Although collectively these parasites infect over a billion people each year, their pathogenic mechanisms have not been completely understood so far. Hence, this review of the literature demonstrates the main mechanisms involved in the pathogenicity of these protozoans, as well as the microenvironmental factors that can interfere with successful colonization. The pathogenesis of E. histolytica involves adhesion, lysis, phagocytosis of epithelial cells and bacteria, tissue invasion by enzymatic action, and evasion of host immune response. Lectin Gal/GalNac, amoebapores, and cysteine proteases are the main molecules involved in these processes. The establishment of giardiosis depends on several pathogenic mechanisms and virulence developed by G. lamblia, such as molecules involved in adhesion, encystation and antigenic variation. For successful colonization of vaginal mucosa, T. vaginalis express molecules like adhesins on the surface and galectin and lipophosphoglycan, involved in the adherence to vaginal epithelial cells and altered gene expression of both the parasite and the host.

Calpains are Involved in Entamoeba histolytica-Induced Death of HT-29 Colonic Epithelial Cells

Entamoeba histolytica is an enteric tissue-invading protozoan parasite that can cause amebic colitis and liver abscess in humans. E. histolytica has the capability to kill colon epithelial cells in vitro; however, information regarding the role of calpain in colon cell death induced by ameba is limited. In this study, we investigated whether calpains are involved in the E. histolytica-induced cell death of HT-29 colonic epithelial cells. When HT-29 cells were co-incubated with E. histolytica, the propidium iodide stained dead cells markedly increased compared to that in HT-29 cells incubated with medium alone. This pro-death effect induced by ameba was effectively blocked by pretreatment of HT-29 cells with the calpain inhibitor, calpeptin. Moreover, knockdown of m- and micro-calpain by siRNA significantly reduced E. histolytica-induced HT-29 cell death. These results suggest that m- and micro-calpain may be involved in colon epithelial cell death induced by E. histolytica.

Mecanismos fisiopatogênicos e diagnóstico laboratorial da infecção causada pela Entamoeba histolytica / Physiopathogenic mechanisms and laboratorial diagnosis of Entamoeba histolytica infection

A amebíase é a segunda causa de morte entre as doenças parasitárias no mundo. Seu agente etiológico é o protozoário Entamoeba histolytica, que através da secreção de proteinases é capazes de destruir o tecido hospedeiro, matando as células-alvo por contato e fagocitando eritrócitos. Dessa forma, os trofozoítos invadem a mucosa intestinal, provocando a colite amebiana. Em alguns casos atravessam a mucosa e, através da circulação porta, chegam ao fígado, onde causam necrose constituída por poucos trofozoítos rodeados de hepatócitos mortos e debris celulares liquefeitos. Essa invasão está diretamente relacionada com a capacidade de síntese e a secreção de moléculas responsáveis pela virulência dos trofozoítos, como os amebaporos, as lectinas e as cisteína proteinases. O diagnóstico da infecção causada pelo patógeno é rotineiramente realizado através da microscopia óptica de amostras frescas ou espécimes fixados. Entretanto essa metodologia apresenta limitações, sendo incapaz de distinguir as espécies pertencentes ao complexo E. histolytica/E. dispar. A pesquisa de coproantígenos e a reação em cadeia da polimerase (PCR) têm sido utilizadas para diferenciação desses protozoários em amostras fecais. No entanto, estudos mais aprofundados são necessários para maior compreensão sobre a relação parasita/hospedeiro, a proteômica e a genômica do protozoário, o desenvolvimento de vacinas e a real prevalência dessa infecção no Brasil e no mundo.

Amebiasis is the second cause of death among parasitary diseases in the world. Its etiologic agent is the protozoan Entamoeba histolytica, which destroys the host tissue by means of the secretion of proteinases, kills the target-cells by contact and phagocytizes erythrocytes. Accordingly, the trophozoites invade the intestinal mucosa, what causes amoebaean colitis. In some cases, they pass through the mucosa and reach the liver through the portal system, where they cause necrosis, which is composed of a few trophozoites surrounded by dead hepatocytes and liquefied cellular debris. This invasion is directly related to the synthesis capacity and secretion of molecules responsible for the virulence of trophozoites such as amoebapores, lectins and cysteine proteinases. The diagnosis of infection caused by this pathogen is routinely performed through optical microscopy of fresh samples or fixed specimens. However this methodology presents limitations insofar as it is unable to distinguish the specimens belonging to the complex E. histolytica /E. dispar. The research on coproantigens and the polymerase chain reaction (PCR) method have been used to differentiate these protozoa in fecal samples. However further studies are required for a better understanding of the host-parasite relationship, the proteomics and genomics of the protozoa, the development of vaccines and the real prevalence of this infection in Brazil and worldwide.

Amebíase e epidemiologia / Amebiasis and epidemiology

A infecção por Entamoeba histolytica, identificada há mais de 130 anos por Fedor A. Lesh, existe praticamente em todo o mundo e é hoje considerada uma DST - com relação à transmissão é a doença dos cinco efes: finger, feces, flies, fomites e fornication. A epidemiologia da amebíase na cidade do Rio de Janeiro (Brasil), estudada por um de nós (R.M.), parece confirmar o lugar de infecção no rol das doenças sexualmente transmissíveis. Epidemiologia é o estudo da ocorrência de uma doença - estudos epidemiológicos podem influenciar a vida de populações inteiras. O estudo de Framingham (EUA), a investigação de Sharr sobre a doença dos legionários e o trabalho de John Snow sobre a cólera são exemplos clássicos de estudos epidemiológicos que mudaram o comportamento e estilos de vida.

Entamoeba histolytica infection was identified more than 130 years ago, has worldwide occurrence and nowadays is considered a sexually transmitted disease (STD). Regarding transmission is considered as the five Fs disease: finger, feces, flies, fomites and fornication. Rio de Janeiro city amebiasis epidemiology was studied by one of us and seems to confirm its place on STD list. Epidemiology consists on the study of the disease occurrence - epidemiological studies can influence an entire population life. Framinghan (EUA), Legionnaire's disease Sharr investigation and John Snow cholera study are classic examples of epidemiological studies that changed behaviour and lifestyle.

Historia del protozoo Entamoeba histolytica: [revisión] / History of the Entamoeba histolytica protozoan: [review]

This article presents a history of Entamoeba histolytica spanning since the remote times when it was not even recognized as a cause of human disease to the recent molecular advances. Feder Losch (1875) in Saint Petersburg, found amoebae in fecal samples but only regarded them as responsible for maintaining the inflammatory process, not as a cause of dysentery. Fritz Schaudinn (1903) established the differentiation between Entamoeba histolytica and Endamoeba coli, Schaudinn decided to call it E. histolytica because of its ability to cause tissue lysis. Emile Brumpt (1925) based on experimental studies, pointed out the existence ofE. Histolytica as a species complex, comprising two morphologically indistinguishable species, E. dysenteríae which is the cause of symptomatic infection, and Entamoeba dispar found only in asymptomatic carriers. Louis Diamond et al (1961) during the 1960s developed an axenic culture medium for E. histolytica which allowed in vivo and in vitro studies. Sargeaunt and Williams (1978) distinguished for the first time E. histolytica strains by isoenzyme electrophoresis, thus confirming thatE. hystolytica was indeed a species complex comprising both pathogenic and non-pathogenic species. William Petri et al (1987 demonstrated that the 170 kDa protein with greater antigenicity was the Gal/GalNac-specific lectin. Diamond and Clark (1993) described again Brumpt's original 1925hypothesis, concluding that there was enough evidence to support the existence of two morphologically indistinguishable species, a pathogenic and a nonpathogenic one, corresponding to E. histolytica and Entamoeba dispar respectively. The World Health Organization accepted this hypothesis in 1997.

Amebíase / Amoebiasis: an update

A amebíase é a segunda principal causa de morte por parasito em todo o mundo. O protozoário responsável, Entamoeba histolytica, apresenta elevada patogenicidade. É capaz de secretar proteases que dissolvem o tecido do hospedeiro, matar suas células por contato, fagocitar eritrócitos e invadir a mucosa intestinal causando a colite amebiana. Em alguns casos, este parasito é capaz de romper a barreira da mucosa intestinal e chegar ao fígado por meio da circulação porta, onde pode causar abscesso que cresce rapidamente e é quase sempre fatal. Evidências baseadas apenas na morfologia apontavam a existência de uma única espécie. No entanto, estudos mais modernos mostraram que, na realidade, há duas espécies geneticamente bem distintas, denominadas Entamoeba histolytica (patogênica) e Entamoeba dispar (não patogênica ou comensal).

Morbidity due to Schistosoma mansoni - Entamoeba histolytica coinfection in hamsters (Mesocricetus auratus)

Data on Schistosoma mansoni-Entamoeba histolytica coinfection are scarce in the literature. In the present study, hamsters that had been infected for 70 days with Schistosoma mansoni (LE strain) were inoculated via the portal vein with two strains of trophozoites of Entamoeba histolytica: ICB-EGG (highly virulent) and ICB-RPS (non-virulent). The most evident result of coinfection was increased morbidity and mortality, in comparison with either of the infections alone. Histologically, there were no evident signs of interaction between these two infections. The morphological findings of schistosomal granuloma and amoebic abscesses in the liver were similar to those seen in the respective single-infection controls. However, there was severe wasting of the animals with both infections and greater numbers of amoebic lesions in their livers. The results obtained indicated that schistosomiasis aggravates the course of amoebiasis in hamsters.

Dados sobre a co-infecção Schistosoma mansoni-Entamoeba histolytica são escassos na literatura. No presente estudo, hamsters com 70 dias de infecção por Schistosoma mansoni (cepa LE) foram inoculados com trofozoítos de Entamoeba histolytica, cepa ICB-EGG (virulenta) e cepa ICB-RPS (não virulenta), via veia porta. O mais evidente resultado da co-infecção foi o aumento da morbidade e mortalidade, quando comparado com os animais com somente uma das infecções. Histologicamente, não houve sinais evidentes da interação entre as duas infecções. O aspecto morfológico do granuloma esquistossomótico e do abcesso hepático amebiano são similares aos observados nos controles, com somente uma infecção. Entretanto, foi observado que os animais co-infectados apresentavam-se mais debilitados e com maior número de lesões amebianas no fígado. Os resultados obtidos indicam que a esquistossomose agrava o curso da infecção amebiana em hamsters.

Participación del óxido nítrico durante el desarrollo del absceso hepático amebiano / Nitric oxide participation during amoebic liver abscess development

El óxido nítrico participa en funciones fisiológicas y fisiopatológicas, así como en el mecanismo de defensa del sistema inmunológico de mamíferos contra parásitos, virus y bacterias. La Entamoeba histolytica es un parásito protozoario causante de la amebiasis, la cual se caracteriza por el daño intestinal y la formación del absceso hepático amebiano (AHA). El desarrollo del absceso hepático amebiano en el hámster es similar al que desarrolla el humano, mientras que el ratón es resistente a la formación de este absceso, debido a un incremento en la producción de óxido nítrico. A diferencia del ratón, el desarrollo del absceso hepático amebiano en el hámster es debido a un exceso en la producción de óxido nítrico o posiblemente a una mayor susceptibilidad del hámster al daño producido por el óxido nítrico. Por lo tanto, sería importante realizar más estudios para determinar si en el humano, un exceso en la producción de óxido nítrico favorece la formación del absceso hepático amebiano.

Nitric oxide participates in both physiological and pathophysiological functions, and it plays an important role in the mammalian immune system in killing or inhibiting the growth of many pathogens, including parasites, viruses and bacteria. Entamoeba histolytica is a protozoan parasite that causes amoebiasis, which is characterized by intestinal damage and amoebic liver abscess development. The development of amoebic liver abscess in hamsters is similar to that in humans, whereas mice are resistant to amoebic liver abscess development due to an increase in nitric oxide production. Unlike in mice, amoebic liver abscess development in hamsters is due to an excess in nitric oxide production or possibly to a greater susceptibility of the hamster to damage caused by nitric oxide. Therefore, it could be important to elucidate if, in humans, an excess in nitric oxide production favors amoebic liver abscess development.

Pathogenicity of Entamoeba dispar under xenic and monoxenic cultivation compared to a virulent E. histolytica / Patogenicidade de Entamoeba dispar em cultivo polixênico e monoxênico comparada a uma cepa virulenta de E. histolytica

Two xenic isolates and cloned cultures of Entamoeba dispar were submitted to monoxenization using Crithidia fasciculata as the associated organism. Growth in monoxenic cultivation and ability of xenic and monoxenic trophozoites to destroy VERO cells and produce lesions in hamster livers were compared to those of a virulent E. histolytica. Parental and cloned E. dispar under monoxenic cultivation showed a remarkable lower growth than the monoxenic E. histolytica and were avirulent in both in vivo and in vitro tests. When xenically cultured, trophozoites of E. dispar showed a moderate lytic activity against VERO cells (1.5 to 41.8 percent of destruction) but caused severe hepatic lesions in hamsters as those caused by the virulent E. histolytica (29 to 100 percent in prevalence and 0.86 to 4.00 in lesion degree). Although E. dispar has not been associated with invasive disease in men, the ability of xenic trophozoites to produce prominent tissue damage in experimental conditions has indicated that some strains have a considerable pathogenic potential when in presence of bacteria.

Dois isolados de Entamoeba dispar em cultivo polixênico e culturas clonadas deles obtidas foram submetidos à monoxenização utilizando Crithidia fasciculata como organismo associado. O crescimento em cultivo monoxênico dos isolados e clones, bem como sua capacidade de destruir células VERO (efeito citopático) e de produzir lesões hepáticas em hamster foram comparados a uma cepa virulenta de E. histolytica. Os trofozoítos de E. dispar em cultivo monoxênico apresentaram um crescimento nitidamente menor que o de E. histolytica e foram avirulentos tanto no teste in vivo quanto in vitro. Entretanto, isolados e clones de E. dispar em cultivo polixênico exibiram uma atividade lítica moderada sobre as células VERO (1,5 to 41,8 por cento de destruição) e causaram lesões hepáticas em hamster (29 a 100 por cento em prevalência e 0,86 a 4,00 no grau de lesão) tão extensas quanto aquelas causadas pela E. histolytica. Embora E. dispar não seja associada à doença invasiva no homem, a ocorrência de lesões teciduais significativas, causadas por trofozoítos em condições experimentais, indica que esta espécie pode apresentar potencial patogênico considerável quando em presença de bactérias intestinais.

Amoebiasis cutis in HIV positive patient.

Protozoan infections of the skin, particularly cutaneous amoebiasis, are rare in HIV-positive patients. We report a case of amoebiasis cutis in an HIV-positive truck driver with a history of frequent unprotected sexual exposures. He presented with multiple painful ulcers and sinuses with purulent discharge, necrotic slough and scarring in the perianal and gluteal region for the last 2 years. He was positive for HIV-1 and -2. Cutaneous biopsy revealed numerous Entamoeba histolytica in the trophozoite form, in addition to an inflammatory infiltrate and necrotic debris. He responded well to oral metronidazole and chloroquine. Amoebiasis cutis should be considered in the differential diagnosis of perianal ulcers, particularly in HIV-positive patients.

[Pathogenesis of intestinal amebiasis]

Amebiasis is the infection of the human gastrointestinal tract by Entamoeba histolytica, a protozoan parasite that is capable of invading the intestinal mucosa and spread to other organs mainly the liver. Identification of proteins associated with virulence, including a lectin that mediates adherence to epithelial cells, a pore-forming peptide that lyses host cells and secreted proteases that degrade host tissues, is one of major recent accomplishment in the field of amebiasis research. This review focuses mainly on the parasitic mechanisms that may be related to invasive intestinal amebiasis

Contact of Entamoeba histolytica with baby hamster kidney-21 (BHK-21) cell line on cysteine proteinase activity.

BACKGROUND & OBJECTIVES: Entamoeba histolytica, the causative agent of amoebiasis and amoebic liver abscess, lyses host cells by direct contact using surface lectins and releases cysteine proteinase (CP). Virulence of E. histolytica is directly related to activity of its CP. The relationship of CP activity and cytotoxicity has not been established. The present study was carried out to explore the events following contact of E. histolytica with target cells. METHODS: Protease activity of E. histolytica was measured by azocaseine and haemoglobin assays, and cysteine proteinase activity was assessed by substrate gel electrophoresis. Target cell lysis was measured by chromium release assay. RESULTS: Protease activity of E. histolytica was increased 2.5-fold following contact with BHK-21 cell line. CP activity of trophozoites alone was visualized at position 56, 35 and 29 kDa in substrate gel electrophoresis. Contact of trophozoites with target cells augmented the cytotoxic activity of amoebic CP. The increase in CP activity seen by substrate gel electrophoresis and cytotoxicity assay was blocked by pretreatment with E 64, a specific CP inhibitor and GalNAc, a contact inhibitor. INTERPRETATION & CONCLUSION: The present data showed the involvement of amoebic CP in cytotoxicity and that the CP activity was enhanced on lectin-mediated contact of E. histolytica to the target cells. Further studies need to be done to understand the mechanism at the molecular level.

Intestinal parasitic infections in children from Northern Pakistan

Intestinal parasitic infections cause significant morbidity in children of developing countries, where the vicious cycle of infections and malnutrition impairs their physical growth and development. A community based cross-sectional study on randomly selected children under 15 years of age was performed in two towns of Northern Pakistan. Information on hygiene, sanitation, waste disposal, water supply and maternal education was collected and the nutritional status of these children was assessed. Stool samples of the subjects were collected, immediately preserved and later examined for parasites, with severity of infection being quantified. Of the 89 samples examined, 91% contained one or more parasites. The lowest prevalence of infection was in children less than one year old. Majority of those infected in both towns were girls. Fifty-one percent of the infected children from Yasin and 38% from Singal suffered from malnutrition. Maternal education, family income and drinking water treatment did not affect the prevalence of infection in either town. The most frequent parasites isolated were Ascaris lumbricoides [66.3%], Entamoeba histolytica [27%], Blastocystis hominis [27%], Giardia lamblia [24.7%], and Trichuris trichiura [15.7%]. The results of this study indicate that parasitic infections in northern Pakistan have a high prevalence but low intensity. Programs for early detection and interventional strategies, along with improved hygiene, sanitation and waste disposal facilities are urgently needed to improve the health of these children

Experimental amoebic liver abcess produced by oral adminsitration of entamoeba histolytica cysts

To determine the possibility of amoebic invasion and liver abscess formation, Swiss albino mice were infected orally with E. Histolytica cysts isolated from human stools. Parasitological and histopathological changes in mice colon and liver tissues were sequentially followed. Three weeks post-infection [pi], 5% of immunocompetent and all cortisonized immunosuppressed mice passed the parasite in their stools. Only 70% of the latter group of mice sacrificed at that time developed invasive intestinal amoebiasis. At the end of the experiment, 100% of the remaining immunosuppressed animals developed the same intestinal pathology. Amoebic liver abscess was detected in 62.5% of them. Oral inoculation of E. histolytica cysts constituted an easy highly reproducible procedure for inducing liver abscess in immunosuppressed mice

Arbitrarily primed PCR fingerprinting of RNA and DNA in Entamoeba histolytica

Differences were detected in the gene expression of strains of E. histolytica using RNA (RAP-PCR) and DNA fingerprinting (RAPD). Analysis of the electrophoretic profiles of the gels revealed some polymorphic markers that could be used in the individual characterization of the strains. The 260 bands generated by using five different primers for RAP-PCR, as well as RAPD, were employed in the construction of dendograms. The dendogram obtained based on the RAPD products permitted the distinction of symptomatic and asymptomatic isolates, as well the correlation between the polymorphism exhibited and the virulence of the strains. The dendogram obtained for the RAP-PCR products did not show a correlation with the virulence of the strains but revealed a high degree of intraspecific transcriptional variability that could be related to other biological features, whether or not these are involved in the pathogenesis of amebiasis