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PubMed1981

Susceptibility of Entamoeba histolytica to oxygen intermediates.

To explore the susceptibility of the extracellular protozoan, Entamoeba histolytica, to toxic oxygen intermediates, trophozoites were exposed to fluxes of O2, H2O2, and OH. generated enzymatically by the glucose oxidase and xanthine oxidase reactions. HM-1 trophozoites were resistant to O2, but were readily killed by H2O2 alone. OH. and 1O2 were not required for effective amebicidal activity. The addition of peroxidase and halide enhanced trophozoite killing by H2O2. Sonicates of amebae contained virtually no catalase and little glutathione peroxidase activity which may contribute to susceptibility to H2O2. Coupled with our previous studies with Toxoplasma gondii and Leishmania spp. these observations indicate that there is a broad spectrum of susceptibility of intra- and extracellular pathogenic protozoa to killing by oxygen intermediates.

Animals↗
PubMed2000

Amebic abscess of the liver presenting as acute cholecystitis.

A case report is presented of a 37-year-old active duty Navy petty officer with amebic abscess of the liver presenting as acute cholecystitis. He was admitted with severe right upper quadrant pain and a positive Murphy's sign, but sonogram and computed tomography (CT) scan demonstrated an abscess in the right lobe of the liver. "Anchovy paste" material was obtained on percutaneous drainage, and he was placed on metronidazole administration with a tentative diagnosis of amebic abscess. This was confirmed on enzyme-linked immunosorbent assay. Symptoms resolved within a few days, and the abscess progressively decreased in size. Amebic abscess of the liver is discussed, with emphasis on pathogenesis, diagnosis, and treatment. Although uncommon, the condition is still seen in various population groups including the United States military. Difficulty in diagnosis is not unusual, and as in the herein-reported case, amebic abscesses of the liver may be confused with acute cholecystitis and other intra-abdominal infections. Abdominal sonogram and CT examination will identify a process in the liver, but the presence of amebiasis must be confirmed by laboratory studies on serum or contents of the abscess. Amebicidal agents are effective in many cases, but there remain roles for aspiration of the abscess, percutaneous drainage, and even open surgical drainage. Failure to establish an early diagnosis may result in rupture of the abscess, with catastrophic results.

Journal Article↗
PubMed2002

Acanthamoeba sclerokeratitis: treatment with systemic immunosuppression.

OBJECTIVE: This study describes the clinical features, management, and outcome of 19 patients who had severe Acanthamoeba sclerokeratitis (ASK) unresponsive to conventional management, requiring systemic immunosuppression to control disease. DESIGN: Retrospective, non-comparative, interventional case series. PARTICIPANTS: Records of all patients with Acanthamoeba keratitis treated at Moorfields Eye Hospital between 1989 and 2000 were reviewed. From more than 200 patients, 19 who developed ASK treated with systemic immunosuppression were identified. MAIN OUTCOME MEASURES: Visual acuity, level of pain, and degree of inflammation were recorded after immunosuppressive treatment. RESULTS: ASK requiring immunosuppression occurred in 20 eyes of 19 patients (11 males and 8 females). The mean age (mean +/- standard deviation) at onset was 38.6 +/- 13.2 years. On presentation, best-corrected visual acuity was counting fingers or worse in 11 eyes (55%), 6/18 to 6/60 in 5 eyes (25%), and 6/12 or better in 4 eyes (20%). The mean time between onset of initial symptoms of Acanthamoeba keratitis and commencement of systemic immunosuppression was 4.8 +/- 3.5 months. The mean duration of immunosuppression required to control inflammation was 7.2 +/- 3.9 months. Severe scleritic pain remained uncontrolled in two patients and resulted in enucleation. Best-corrected visual acuity at final follow-up was counting fingers or worse in eight eyes (40%), 6/18 to 6/60 in six eyes (30%), and 20/40 or better in six eyes (30%). The mean follow-up period after resolution of inflammation was 24.3 +/- 20.9 months (range, 0.2-59.7 months). CONCLUSIONS: ASK is an uncommon complication of Acanthamoeba keratitis. The scleritis associated with this infection seems to be an immune-mediated response. After topical amebicidal treatment, systemic immunosuppression may be required to control the pain and tissue destruction associated with ASK.

Acanthamoeba Keratitis↗
PubMed1990

[Effects of Saccharomyces boulardii yeast on trophozoites of Entamoeba histolytica in vitro and in cecal amebiasis in young rats].

Lyophilized and rehydrated Saccharomyces boulardii yeasts were administered to young rats previously inoculated into the cecum with Entamoeba histolytica. The numbers of diseased young rats and the severity of the infection, assessed from the appearance of the cecum and its content of mucus and amebae, were significantly reduced by the treatment. The lesions observed resembled those seen in the untreated controls, although healing was faster in the treated animals. Saccharomyces boulardii had no intrinsic amebicidal action in vitro.

Animals↗
PubMed2003

Identification and characterization of two isoenzymes of methionine gamma-lyase from Entamoeba histolytica: a key enzyme of sulfur-amino acid degradation in an anaerobic parasitic protist that lacks forward and reverse trans-sulfuration pathways.

To better understand the metabolism of sulfur-containing amino acids, which likely plays a key role in a variety of cell functions, in Entamoeba histolytica, we searched the genome data base for genes encoding putative orthologs of enzymes known to be involved in the metabolism. The search revealed that E. histolytica possesses only incomplete cysteine-methionine conversion pathways in both directions. Instead, this parasite possesses genes encoding two isoenzymes of methionine gamma-lyase (EC 4.4.1.11, EhMGL1/2), which has been implicated in the degradation of sulfur-containing amino acids. The two amebic MGL isoenzymes, showing 69% identity to each other, encode 389- and 392-amino acid polypeptides with predicted molecular masses of 42.3 and 42.7 kDa and pIs of 6.01 and 6.63, respectively. Amino acid comparison and phylogenetic analysis suggested that these amebic MGLs are likely to have been horizontally transferred from the Archaea, whereas an MGL from another anaerobic protist Trichomonas vaginalis has MGL isotypes that share a common ancestor with bacteria. Enzymological and immunoblot analyses of the partially purified native amebic MGL confirmed that both of the MGL isotypes are expressed in a comparable amount predominantly in the cytosol and form a homotetramer. Recombinant EhMGL1 and 2 proteins catalyzed degradation of L-methionine, DL-homocysteine, L-cysteine, and O-acetyl-L-serine to form alpha-keto acid, ammonia, and hydrogen sulfide or methanethiol, whereas activity toward cystathionine was negligible. These two isoenzymes showed notable differences in substrate specificity and pH optimum. In addition, we showed that EhMGL is an ideal target for the development of new chemotherapeutic agents against amebiasis by demonstrating an amebicidal effect of the methionine analog trifluoromethionine on trophozoites in culture (IC50 18 mum) and that this effect of trifluoromethionine was completely abolished by the addition of the MGL-specific inhibitor DL-propargylglycine.

Amino Acids, Sulfur↗
PubMedDate not supplied

Review of the human immune mechanisms directed against Entamoeba histolytica.

The human immune mechanisms effective against the destructive parasite Entamoeba histolytica have not been established. Humoral responses develop with invasive disease but have not positively correlated with protection against recurrent invasive infection. Though complement is amebicidal, invasive strains of amebas may be resistant. Polymorphonuclear neutrophils have not been active in vitro against virulent amebic strains and could contribute to the pathogenesis of disease. In vitro studies have shown human activated monocyte-derived macrophages and cytotoxic T lymphocytes to be competent cells in killing virulent amebas. Soluble amebic protein preparations have been shown to be mitogenic for normal, uninfected human lymphocytes, apparently because the N-acetyl-D-galactosamine-inhibitable amebic lectin is present. Despite a lower T4:T8 ratio and a heterogeneous lymphocyte proliferation in response to mitogens, patients with amebic hepatic abscess develop sensitization to amebic antigens and specific effector mechanisms. Although antibody, complement, and nonimmune mechanisms, as well as parasite factors, may be important in determining the occurrence of invasive amebiasis, human cell-mediated immune mechanisms may play a major role in combating invasive infection due to E. histolytica.

Animals↗
PubMed1989

Entamoeba histolytica: from adherence to enteropathy.

Entamoeba histolytica, a cause of invasive colitis or liver abscess, is responsible for substantial worldwide morbidity and mortality. An understanding of the biochemical basis for the parasite adherence and cytolytic activities and antiamebic host immune response mechanisms are prerequisites for vaccine development. The E. histolytica galactose (Gal) or N-acetyl-D-galactosamine (GalNAc) inhibitable adherence lectin mediates attachment of trophozoites to colonic mucins or mammalian target cells. Amebic cytolysis of target cells requires Gal/GalNAc-lectin-mediated adherence, parasite phospholipase A activity, and maintenance of an acid pH in amebic intracellular vesicles. Cytolytic activity is stimulated by phorbol esters (activators of protein kinase C) and results from an E. histolytica-mediated increase in free Ca++ within the target cell. The Gal/GalNAc adherence lectin is a highly conserved antigen that is universally recognized by human immune sera; patients cured of invasive amebiasis possess antigen-specific cell-mediated immunity effective in vitro against E. histolytica trophozoites. Promising vaccines include the purified adherence lectin, for eliciting an intestinal secretion of IgA antibody to lectin, and additional E. histolytica antigens, which elicit cell-mediated amebicidal responses.

Amebiasis↗
PubMed1987

A comparison of amebic and pyogenic abscess of the liver.

We evaluated the clinical features of 96 cases of amebic liver abscess and 48 of pyogenic hepatic abscess. Most patients with amebic abscess were young Hispanic males. Those with pyogenic abscess were older, without any ethnic predominance. Symptoms tended to be acute and localized to the right upper quadrant in amebic infection. In pyogenic disease, symptoms were often nonspecific and chronic in nature. A marked shift to the left of the leukocyte count occurred more frequently in pyogenic abscess, as did markedly abnormal values of the serum albumin, direct bilirubin, lactic dehydrogenase and aspartate aminotransferase. Sonography detected all cases of amebic abscess and missed the lesions in 2 of 39 patients with pyogenic abscess. Abscess cultures yielded pathogens in 90% of cases of pyogenic disease, while blood cultures were positive in 50%. Five of 20 patients with positive blood cultures had additional organisms isolated from the abscess that would have required adjustment of antibiotics for optimal coverage. We believe that all pyogenic abscesses should be aspirated to guide antibiotic therapy. In amebic abscess, the diagnosis was usually based on clinical and sonographic findings, aspiration being performed in only 14% of cases. Ninety-eight percent of patients were treated with amebicidal agents alone, and all responded to therapy. Therapeutic needle aspiration is rarely necessary. In pyogenic abscess, prolonged fever was common during medical therapy. Even in those eventually cured without surgery, the median time to defervescence was 8 days. Though 19 patients underwent surgical drainage, only 2 clearly did not benefit from medical treatment, having high fevers after more than 2 weeks on a regimen of appropriate antibiotics. Surgery is often performed prematurely because physicians expect fever to resolve quickly, but persistent fever of less than 2 weeks' duration should not constitute an indication for surgical drainage. Seven patients with pyogenic abscess died, 5 as a result of hepatic abscess. In 3 of these cases, the diagnosis was unsuspected till autopsy. Improved awareness of this disease may decrease morbidity and mortality from this treatable condition.

Adult↗
PubMed1994

Perforated amebic liver abscess: clinical analysis of 110 cases.

In this study of 503 consecutive cases of amebic liver abscess documented during a 21-year period, 110 (22%) were complicated by perforation. Perforation sites included pleuropulmonary structures (79 cases, 72%) subphrenic space (15 cases, 14%); and peritoneal cavity (11 cases, 10%). The less common sites were the thoracic and abdominal wall, pericardial sac, bile duct, kidney, mediastinum, colon, and right flank. Trophozoites were recovered in 29% of cases. For the rest of the cases, diagnosis was based on the clinical manifestations and the dramatic response to a therapeutic trial of amebicides. Open surgical drainage was done in 10 of 11 patients who had peritoneal perforation; 6 of them were cured and 5 died. Of 97 patients with perforation sites other than the peritoneal cavity, 64 were treated with metronidazole in addition to needle aspiration and/or percutaneous catheter drainage, and all achieved complete remission without relapse during subsequent follow-up. Of the 36 cases treated with metronidazole alone, 22 (61%) were cured. The mortality rate in the group with perforated abscess was 17% (19/110), compared with 5% (25/503) in the whole series. The results suggested that (1) amebic liver abscess can be difficult to diagnose, especially when the pathogen is not isolated; and (2) metronidazole combined with needle aspiration and/or percutaneous catheter drainage is the treatment of choice for amebic liver abscess with perforation.

Adolescent↗
PubMedDate not supplied

Efficacy of novel antimicrobials against clinical isolates of opportunistic amebas.

We examined the effects of the macrolide antimicrobial agent azithromycin and phenothiazine compounds against clinical isolates of Acanthamoeba spp. and Balamuthia mandrillaris, opportunistic pathogens of human beings and other animals. Acanthamoeba growth was inhibited in vitro at 1, 5, and 10 micrograms/ml of azithromycin, but not the macrolides, erythromycin, and clarithromycin. In experiments attempting to simulate in vivo conditions, azithromycin protected monolayers of rat glioma cells from destruction by Acanthamoeba at a concentration of 0.1 microgram/ml, and delayed destruction at concentrations of 0.001 and 0.01 microgram/ml. We concluded that the minimal inhibitory concentration of azithromycin was 0.1 microgram/ml. Our results, however, suggested that the drug was amebastatic but not amebicidal, since ameba growth eventually resumed after drug removal. The phenothiazines (chlorpromazine, chlorprothixene, and triflupromazine) inhibited Acanthamoeba growth by 70-90% at 5 and 10 micrograms/ml, but some of these compounds were toxic for rat glioma cells at 10 micrograms/ml. Azithromycin was not very effective against B. mandrillaris in an in vitro setting, but was amebastatic in tissue culture monolayers at concentrations of 0.1 microgram/ml and higher. Balamuthia amebas showed in vitro sensitivity to phenothiazines. Ameba growth was inhibited 30-45% at 5 micrograms/ml in vitro, but completely at 5 micrograms/ml in the rat glioma model. In spite of their potential as antiamebic drugs in Balamuthia infections, toxicity of phenothiazines limits their use in clinical settings.

Acanthamoeba↗
PubMedDate not supplied

The interaction of Acanthamoeba spp. with activated macrophages and with macrophage cell lines.

Acanthamoeba spp. are free-living amebae associated with amebic keratitis and chronic granulomatous amebic encephalitis. The present studies were undertaken to compare the pathogenicity of three species of Acanthamoeba in B6C3F1 mice after intranasal challenge with Acanthamoeba-induced cytopathogenicity for different macrophage populations. The ability of murine macrophage cell lines and activated murine peritoneal macrophages to lyse Acanthamoeba has been assessed by coincubating macrophages with 3H-uridine labeled amebae. Conversely, destruction of macrophages by Acanthamoeba was determined by measuring the release of chromium-51 from radiolabeled macrophages. Acanthamoeba culbertsoni, which is highly pathogenic for mice, destroys macrophage cultures in vitro. Activated primary peritoneal macrophages were more resistant to Acanthamoeba-mediated destruction than macrophage cell lines activated in vitro. Activated macrophages were capable of limited destruction of Acanthamoeba polyphaga and Acanthamoeba castellanii. Acanthamoeba-specific antibodies increased the amebicidal activity of activated macrophages. Macrophage-mediated destruction was by contact-dependent cytolysis and by ingestion of amebae. Conditioned medium obtained from macrophage cultures after treatment with lipopolysaccharide and interferon gamma was neither cytolytic nor cytostatic for Acanthamoeba spp. Purified recombinant cytokines including tumor necrosis factor alpha, interleukin 1 alpha, and interleukin 1 beta, alone or in combination, were not cytolytic for Acanthamoeba trophozoites.

Acanthamoeba↗
PubMed1994

Susceptibility testing of Dientamoeba fragilis ATCC 30948 with iodoquinol, paromomycin, tetracycline, and metronidazole.

Susceptibility testing was performed on Dientamoeba fragilis ATCC 30948 in a dixenic culture with Klebsiella pneumoniae and Bacteroides vulgatus. D. fragilis was cocultured with the bacteria in TYGM-9 medium (ATCC medium 1171). The activities of antiparasitic drugs were assessed by counting viable D. fragilis trophozoites with a hemacytometer by trypan blue exclusion. The minimal amebicidal concentrations of the following four drugs were determined: iodoquinol at 128 micrograms/ml, paromomycin at 16 micrograms/ml, tetracycline (questionably) at 32 micrograms/ml, and metronidazole at 32 micrograms/ml.

Animals↗
PubMed1995

Susceptibility of Acanthamoeba castellanii to contact lens disinfecting solutions.

A corneal isolate of Acanthamoeba castellanii was exposed to commercial contact lens disinfecting solutions containing hydrogen peroxide, benzalkonium chloride, polyaminopropyl biguanide, polyquaternium 1, and chlorhexidine-thimerosal. The minimum trophozoite amebicidal concentration and exposure times required to kill trophozoites and cysts were determined. Solutions containing hydrogen peroxide or chlorhexidine-thimerosal were active against both trophozoites and cysts. The benzalkonium chloride-based solution was effective only against trophozoites. Solutions containing polyaminopropyl biguanide or polyquaternium 1 were completely ineffective. The need for adequate exposure times must be stressed.

Acanthamoeba↗
PubMed1974

In vitro and in vivo activity of 5-fluorocytosine on Acanthamoeba.

The results of our studies indicated that the avirulent Neff strain of Acanthamoeba was more susceptible to the activity of the anti-metabolite 5-fluorocytosine (5-FC) than was the virulent A-1 strain or a mouse brain reisolate of this strain, designated A-3. Results of competition experiments in which cultures were exposed simultaneously to 5-FC and either uracil, thymidine, or both uracil and thymidine demonstrated that the drug was directed against both deoxyribonucleic acid and ribonucleic acid in the avirulent strain, whereas ribonucleic acid was mainly affected in the virulent amebas. Concentrations >10 mug of 5-FC per ml were amebicidal to the avirulent strain; lower concentrations of the drug, which only affected growth slightly, significantly impaired the capacity of the cells to spontaneously encyst in stationary-phase cultures. On the other hand, the virulent strains were capable of growing in the presence of 5-FC (40 mug/ml) after an initial period of susceptibility. After a few transfers in growth medium lacking the drug, 5-FC-treated virulent amebas exhibited growth parameters typical of untreated cells. However, after successive subcultures in drug-free medium, 5-FC-treated cells lost their resistance and were again susceptible to the drug. This result suggested that the capacity of the cells to develop resistance resulted from a drug-induced mechanism. Spontaneous encystment, which was normally minimal in stationary-phase A-1 or A-3 cultures, was enhanced in A-3 but not A-1 cultures treated with 5-FC (>30 mug/ml). Results obtained from experiments to determine the effectiveness of 5-FC in protecting mice experimentally infected with either A-1 or A-3 amebas indicated that the clinical usefulness of 5-FC may be limited by the capacity of the amebas to develop resistance.

Acanthamoeba↗
PubMed1993

Inhibition of Acanthamoeba species by Pseudomonas aeruginosa: rationale for their selective exclusion in corneal ulcers and contact lens care systems.

Cocultivation of Acanthamoeba castellanii and Acanthamoeba polyphaga with live Pseudomonas aeruginosa and with broth filtrates of P. aeruginosa proved equally lethal to the Acanthamoeba spp. The P. aeruginosa-induced amebicidal activity is apparently toxin mediated and has two operative modes: it can function through binding of P. aeruginosa to the ameba membrane and in the presence of one or more P. aeruginosa exoproducts.

Acanthamoeba↗
PubMed1985

Interaction of human leukocytes and Entamoeba histolytica. Killing of virulent amebae by the activated macrophage.

Capable effector mechanisms in the human immune response against the cytolytic, protozoan parasite Entamoeba histolytica have not been described. To identify a competent human effector cell, we studied the in vitro interactions of normal human polymorphonuclear neutrophils, peripheral blood mononuclear cells (PBMC), monocytes (MC), and MC-derived macrophages with virulent axenic amebae (strain HMI-IMSS). Amebae killed neutrophils, PBMC, MC, and MC-derived macrophages (P less than 0.001), without loss of parasite viability. The addition of heat-inactivated immune serum did not enable leukocytes to kill amebae, nor did it protect these host cells from amebae. MC-derived macrophages, activated with lymphokine elicited by the mitogens conconavalin A, phytohemagglutinin, or an amebic soluble protein preparation (strain HK9), killed 55% of amebae by 3 h in a trypan blue exclusion assay (P less than 0.001); during this time, 40% of the activated macrophages died. Lysis of amebae was confirmed using 111Indium oxine radiolabeled parasites and was antibody independent. Macrophage death appeared to be due to the deleterious effect of lysed amebae rather than the contact-dependent effector mechanisms of E. histolytica. Adherence between activated macrophages and amebae was greater than that between other leukocytes and amebae (P less than 0.001). Microscopic observations, kinetic analysis of the killing of amebae by activated macrophages, and suspension of amebae with adherent activated macrophages in a 10% dextran solution indicated that contact by activated macrophages was necessary to initiate the killing of amebae. Catalase but not superoxide dismutase inhibited the amebicidal capacity of activated macrophages (P less than 0.001). However, activated macrophages from an individual with chronic granulomatous disease were able to kill amebae, but not as effectively as normal cells (P less than 0.01). In summary, activated MC-derived macrophages killed virulent E. histolytica trophozoites through a contact-dependent, antibody-independent mechanism involving oxidative-dependent and -independent processes.

Adult↗
PubMed1984

Metabolism of 14C-iodochlorhydroxyquin in the dog and the rat.

The disposition and metabolism of iodochlorhydroxyquin (clioquinol), an amebicidal drug with neurotoxic properties, were studied in dogs and rats with 14C-labelled drug. Pharmacokinetic studies in the dog demonstrated that the compound was well absorbed; the bioavailability was 36% of the dose of 1 mg/kg. The serum half-life was 1.3-1.8 h. In both the dog and the rat, biliary excretion was a major route of elimination. The dog excreted 27% of an intravenously administered dose (1 mg/kg) in the bile within 2 h; the rat excreted 39% of the dose (5 mg/kg i.v.) in less than 3 h. Elimination via the renal route was also substantial in both species. Urinary and biliary metabolites were separated by TLC (thin layer chromatography) and identified as sulfate and glucuronide conjugates in both species. No evidence for any other metabolites was found. A significant difference was observed between the dog and the rat in the extent of conjugation; the percentage radioactivity in the urine accounted for by the unchanged compound was six to twenty times greater for the dog than for the rat. The species differences in the disposition and metabolism of the compound might explain its greater toxicity in the dog than in the rat.

Administration, Oral↗
PubMed2003

Hepatic amebiasis.

Amebiasis can be considered the most aggressive disease of the human intestine, responsible in its invasive form for clinical syndromes, ranging from the classic dysentery of acute colitis to extra-intestinal disease, with emphasis on hepatic amebiasis, unsuitably named amebic liver abscess. Found worldwide, with a high incidence in India, tropical regions of Africa, Mexico and other areas of Central America, it has been frequently reported in Amazonia. The trophozoite reaches the liver through the portal system, provoking enzymatic focal necrosis of hepatocytes and multiple micro-abscesses that coalesce to develop a single lesion whose central cavity contains a homogeneous thick liquid, with typically reddish brown and yellow color similar to "anchovy paste". Right upper quadrant pain, fever and hepatomegaly are the predominant symptoms of hepatic amebiasis. Jaundice is reported in cases with multiple lesions or a very large abscess, and it affects the prognosis adversely. Besides chest radiography, ultrasonography and computerized tomography have brought remarkable contributions to the diagnosis of hepatic abscesses. The conclusive diagnosis is made however by the finding of Entamoeba histolytica trophozoites in the pus and by the detection of serum antibodies to the amoeba. During the evolution of hepatic amebiasis, in spite of the availability of highly effective drugs, some important complications may occur with regularity and are a result of local perforation with extension into the pleural and pericardium cavities, causing pulmonary abscesses and purulent pericarditis, respectively The ruptures into the abdominal cavity may lead to subphrenic abscesses and peritonitis. The treatment of hepatic amebiasis is made by medical therapy, with metronidazole as the initial drug, followed by a luminal amebicide. In patients with large abscesses, showing signs of imminent rupture, and especially those who do not respond to medical treatment, a percutaneous drainage must be performed with either ultrasound or computerized tomography guidance. Surgical drainage by laparotomy is reserved to patients with secondary infections.

Animals↗