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PubMed1989

Nondysenteric intestinal amebiasis. Colonic morphology and search for Entamoeba histolytica adherence and invasion.

There is controversy regarding the presence of colonic mucosal abnormalities or mucosal invasion by Entamoeba histolytica in patients with "nondysenteric intestinal amebiasis." To determine the role of E. histolytica in causing symptoms and mucosal changes and to detect if mucosal invasion by E. histolytica is present in nondynsenteric intestinal amebiasis, we evaluated 24 E. histolytica-infected patients (stool microscopy positive for E. histolytica) and 12 noninfected controls who presented with chronic gastrointestinal symptoms, but without dysentery, to a clinic in Calcutta. The colonic mucosa was evaluated at colonoscopy, and mucosal biopsies obtained from the cecum, sigmoid colon, and rectum were evaluated by light microscopy, indirect immunofluorescence microscopy, and scanning electron microscopy. At colonoscopy mucosal ulcerations were absent in all the controls and all except one of the E. histolytica-infected patients. E. histolytica trophozoites or cysts were not seen in the lamina propria or on the luminal surface in any infected patient by light and immunofluorescence microscopy. On scanning electron microscopy, structures that resembled rounded E. histolytica trophozoites were seen on the luminal surface in two of 19 cecal specimens from the infected patients. Moderate or severe mucosal inflammation was frequent on light microscopy in both the E. histolytica-infected patients and the noninfected controls with the cecum involved in two thirds of both groups. Antibodies to E. histolytica were detected in serum of 25% of study patients and 58% of controls. Mucosal inflammation did not correlate with stool positivity for E. histolytica or seropositivity for ameba antibody.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Appendiceal infection by Entamoeba histolytica and Strongyloides stercoralis presenting like acute appendicitis.

A 58-year-old male from Puerto Rico who was taking orally administered cortisone analogs for chronic obstructive pulmonary disease presented with fever, absolute eosinophilia, right lower quadrant pain, and rebound tenderness associated with Strongyloides stercoralis infection of the appendix. A 37-year-old alcoholic male developed fever, right lower quadrant abdominal pain, and rebound tenderness because of infection of the appendix with Entamoeba histolytica. These are the seventh reported case of isolated amebic appendicitis and the ninth reported case of appendiceal involvement with Strongyloides. In all these cases the diagnosis was made only after surgery. Patients with unexplained right lower quadrant pain, particularly if immunosuppressed or with an appropriate travel history, should have stool examinations for ova and parasites. Early diagnosis and treatment may prevent life-threatening complications such as perforation and peritonitis.

Adult↗
PubMed1975

Pathogenicity of entamoeba histolytica.

The pathogenicity of Entamoeba histolytica is discussed from an immunologic point of view. The evidence that there is some "trigger" mechanism which converts a commensal dwelling organism into a tissue invasive pathogen is rejected as inadequate. The number of liver abscess cases in comparison with the number of intestinal amebic infections in a population is so low that this in itself suggests that tissue invasion is a rare event in the life history of the ameba. A review is made of the experimental evidence that some type of sensitization is necessary before ameba can invade tissue. In postulating an immunologic basis for the pathogenicity of ameba, a parallel between the behavior of malignant cells in the body and an amebic infection in the gut is made. An appealing hypothesis which deserves further research effort is that an altered immune response is the basis for the pathogenic mechanism in the host.

Antibody Formation↗
PubMedDate not supplied

Characterization of 20 Entamoeba histolytica strains isolated from patients with HIV infection.

Twenty Entamoeba histolytica strains from patients with HIV-1 infection were isolated and compared with E. histolytica strains from patients without HIV infection. The isoenzyme pattern of the hexokinase as well as the hybridization with known DNA-probes were used as markers for pathogenicity. According to these markers, all 20 strains could be regarded as being nonpathogenic. Direct measurements of the virulence were carried out: destruction of monolayer tissue culture cells, capacity of phagocytosis and the ability to induce liver abscesses in the hamster. The virulence of strains from HIV patients was comparable to that of E. histolytica strains which had been isolated from HIV-negative asymptomatic carriers. In agreement with this, none of the HIV-positive patients showed symptoms of an invasive amebiasis. By PRC, no HIV-1 proviral DNA could be evidenced in the E. histolytica strains which had been isolated from HIV patients.

Acquired Immunodeficiency Syndrome↗
PubMedDate not supplied

Inhibition of target cell adhesion by monoclonal antibody to the 66 kD surface antigen of Entamoeba histolytica.

The adherence of Entamoeba histolytica trophozoites to target cells was studied using a monoclonal antibody to a major surface antigen of 66 kD. Preincubation of trophozoites with monoclonal antibody decreased their ability to adhere to and engulf erythrocytes and to destroy Chinese hamster ovary cells. The monoclonal antibody was specific for the 66 kD antigen, which is possibly a major participant in the adhesion that precedes phagocytosis and cytopathic effects.

Animals↗
PubMed1988

Amebic liver abscess in a European patient: zymodeme classification of Entamoeba histolytica.

This is a case report of a 36-year-old patient who developed an amebic liver abscess after a stay in the Sudan. He was first misdiagnosed as having pneumonia of the right lower lobe. Following establishment of the correct diagnosis, the patient recovered fully after metronidazole treatment. The fecal culture in Robinson's medium yielded extensive growth of Entamoeba histolytica. Electrophoretic characterization proved it to be a zymodeme XIX, which is one of the zymodemes associated with pathogenicity in the host. This first report of a zymodeme classification of E. histolytica in Germany should initiate further epidemiological studies.

Adult↗
PubMedDate not supplied

Isoenzyme analysis of Entamoeba histolytica for evaluation of pathogenicity: detection of zymodeme XIX in South America.

Authors report the results of the isoenzyme analysis of strains of Entamoeba histolytica isolated from international travellers. This recently developed method allows the detection of pathogenic strains by evaluating the electrophoretic mobility of cytoplasmic enzymes and was proved to be more reliable and quickly feasible than previous ones. The experience reported refers to three strains isolated from travellers coming from Ecuador, Brazil and Indonesia, respectively; the zymodeme XIX (in accordance with the Sargeaunt's classification) was identified in all the three cases. This zymodeme has been first detected in 1981 and should currently be considered rare; moreover, it has never been previously reported from the Americas.

Amebiasis↗
PubMed1998

Synergy between Entamoeba histolytica and Escherichia coli in the induction of cytokine gene expression in human colon epithelial cells.

Entamoeba histolytica induces the expression and secretion of interleukin 8 (IL-8) by cultured colon epithelial cells. We assessed the array of cytokine genes expressed by human colon epithelial cells in response to co-culture with E. histolytica trophozoites and tested the hypothesis that enteric bacteria may alter the E. histolytica-induced expression of such genes. HT-29 colon epithelial cells were co-cultured with E. histolytica trophozoites in the presence or absence of Escherichia coli. Cytokine gene expression was assessed by quantitative reverse-transcription polymerase chain reaction (RT-PCR). IL-8 mRNA in colon epithelial cells was up-regulated following exposure to E. histolytica and this was paralleled by increased IL-8 secretion. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-1alpha/beta mRNAs were also up-regulated in these cells. When HT-29 cells were co-cultured with E. coli DH5alpha and E. histolytica there was a synergistic increase in the expression of IL-8, IL-1alpha, and GM-CSF. These results suggest that enteric bacteria may significantly affect early proinflammatory signals produced in host tissues in response to E. histolytica infection.

Animals↗
PubMed2000

Auxotrophy to lipoproteins of Entamoeba histolytica cultivated under axenic conditions.

Entamoeba histolytica grows in media without serum but with a mixture of aminoacids, vitamins, lipoproteins, free cholesterol, phospholipids and fatty acids called PACSR. The ability of lipoproteins and free lipids to support growth of three E. histolytica strains (HK9, HMI:IMSS and HM3:IMSS) was analysed. Tubes containing 5 ml culture medium, amino acids, vitamins and either 120-1,200 microg lipoproteins/ml or 0.017-0.10 mg free lipids/ml (predissolved in absolute ethanol) were inoculated with 1x10(4) trophozoites/ml and incubated at 37 degrees C for 72 h. Amoebae died within 12 h in the presence of any free lipid combination, while those having 240-480 mg lipoproteins/ml reached densities similar to or higher than those of controls (depending on strain). The addition of ethanol (0.1%) to the media produced stable lipid solutions and did not show significant adverse effects. Accordingly, E. histolytica is auxotrophic to lipoproteins and unable to use free cholesterol, phospholipids or fatty acids.

Animals↗
PubMed2000

Interaction of antibodies with Entamoeba histolytica trophozoites from experimental amebic liver abscess: an immunocytochemical study.

Using immunocytochemical techniques, we studied the interaction of antibodies with Entamoeba histolytica trophozoites present during the development of amebic liver abscess. Hamsters were intrahepatically inoculated with HM1-IMSS axenic amebas and sacrificed at different days post-inoculation. IgG of rabbit anti-E. histolytica and IgG of rabbit anti-IgG of hamster were used, both labeled with peroxidase. With the rabbit anti-E. histolytica, all trophozoites present in hepatic lesions from 1-7 days post-inoculation were highly labeled. The IgG of rabbit anti-IgG of hamster intensively stained only those trophozoites present in lesions from 1-2 days post-inoculation. From day 3, the intensity and number of labeled trophozoites decreased progressively. The results suggest that the interaction between the amebas and the IgG of hamster is non-specific during the first 2 days. The absence of labeling in the chronic stages could be due to changes in the membrane antigens of the parasite or to alterations in the bloodstream around necrosis. Also, the anti-E. histolytica antibodies produced in the serum during the development of the hepatic disease are apparently incapable of reaching and interacting with the trophozoites present on the liver abscess. This can explain in part why antibodies do not have an important role in the defense of the host.

Animals↗
PubMed2000

Direct amplification of Entamoeba histolytica DNA from amoebic liver abscess pus using polymerase chain reaction.

An important and serious complication of intestinal infection with Entamoeba histolytica is the involvement of the liver (hepatic amoebiasis). Hepatic amoebiasis is usually diagnosed by the clinical picture (pain in the right upper quadrant and fever), ultrasound examination and positive serology. However, none of these tests are definitive and the picture overlaps with pyogenic liver abscess caused by bacteria. It is for this reason that the feasibility of using polymerase chain reaction (PCR) for the detection of E. histolytica DNA in liver abscess pus was investigated. A comparative study was done to verify the sensitivity of ten pairs of primers specific for detecting E. histolytica in stools. Samples of liver abscess pus from 22 serology-positive patients were collected under ultrasound guidance; and these were used directly in PCR assays without any prior pre-treatment of the samples. Of the ten pairs of previously published primers tested, two pairs of primers (PI + P2 and P11 + P12) were found to give 100% sensitivity. Based on these results, we recommend that PCR assay can be successfully used to confirm the diagnosis of amoebic liver abscess with the primers identified.

Animals↗
PubMed1996

Entamoeba histolytica: gene linkage groups and relevant features of its karyotype.

We identified some gene linkage groups in Entamoeba histolytica using a 4-M urea improved transversal alternating field electrophoresis (TAFE) method. Complex rosette-structured DNA molecules were found trapped along the gel lanes, explaining the fuzziness of the patterns. Using several episomal probes, including 16 S, 5.8 S, and 25 S ribosomal (r)Dna genes, an autonomous replication sequence (ARS), and EhVR1, we identified a complete ribosomal episome linkage group (CELG) at the 1.2-Mb position. Three other incomplete groups were found: IELG-1, formed by EhVR1, 16 S, 5.8 S, and 25 S genes; IELG-2 formed by EhVR1, 16 S and 25 S; and IELG-3 formed only by 5.8 S. Ehadh3, Ehpfo, and Ehredox genes migrated at the 1.8-Mb position, forming the non-ribosomal linkage group, NRLG-1.8, while the Ehenl-1 gene migrated at 1.6 Mb forming the NRLG-1.6 group. Ehhk was located at 1.2, 0.8, and 0.17 Mb in three different groups: NRLG-1.2, IELG-3-0.8, and NRLG-0.17. Putative lineal chromosomes were also identified using an heterologous telomeric probe. By in situ hybridization experiments, the rDNA and Ehhk genes were located in both nucleus and cytoplasm, while the Ehpfo and Ehredox genes were found mainly in the nucleus. We propose a model hypothezising that the 16 S and 25 S genes are in a linear molecule, duplicated in two inverted repeats, which may be looped out of the linear DNA to form an episome probably lacking or not the 5.8 S sequence, which could be added later by recombination.

Animals↗
PubMed2000

Circular and linear DNA molecules in the Entamoeba histolytica complex molecular karyotype.

Entamoeba histolytica genome was analysed by pulsed field gel electrophoresis under conditions to separate linear chromosomes in the 170-1400 kb range. We identified linear DNA molecules of 227, 366, 631, 850, 1112 and 1361 kb (mean sizes obtained by three different methods) and we estimated their reorientation times and migration velocities at various experimental conditions. DNA shift mobility assays, using ethidium bromide, suggested that bands migrating at 227 and 631 kb contain linear and circular DNA, whereas a band at 436 kb has only circular DNA. We obtained a regression equation relating sizes of supercoiled DNA molecules with their migration velocities during a pulse at constant electric field and temperature. We also developed a computer program (EHPATTERNS) that predicts the migration per pulse and the resolution order of circular and linear E. histolytica DNA at different pulse times and constant driving and frictional forces. The simulation showed that linear DNA molecules frequently co-migrate with circular molecules, but circular molecules change when the pulse time varies. This molecular mixture generates broad bands and difficulties in the interpretation of the molecular karyotype of E. histolytica.

Animals↗
PubMed2005

Expression of inducible nitric oxide synthase mRNA and nitric oxide production during the development of liver abscess in hamster inoculated with Entamoeba histolytica.

The present study analyzed iNOS and eNOS mRNA expression and NO production during development of hepatic abscess caused by Entamoeba histolytica trophozoites. One 374-bp sequence, which displayed 88% identity to mammalian iNOS protein, was isolated from LPS-stimulated peritoneal hamster macrophages. A separate 365-bp cDNA sequence showed 99% identity with eNOS protein. iNOS mRNA was detected in hamsters during formation of amoebic liver abscesses, but not in control hamsters. eNOS mRNA expression was not modified. Serum nitrite concentration in hamsters infected with E. histolytica was 33 +/- 6 microM, in control hamsters was 20 +/- 3 microM. The study shows that iNOS mRNA expression and NO production are induced by E. histolytica trophozoites during amoebic liver abscess formation. However, in spite of iNOS mRNA expression and NO production, E. histolytica trophozoites induced liver abscess formation in hamster.

Amino Acid Sequence↗
PubMed2005

MonoADP-ribosylation of the NAD+-dependent alcohol dehydrogenase from Entamoeba histolytica.

The human parasite Entamoeba histolytica is an amitochondrial protozoan whose metabolism depends on glucose fermentation. Among the metabolic enzymes absolutely required for amoeba growth is the NAD+-dependent alcohol dehydrogenase (EhADH2). The polymeric form of EhADH2 was sedimented at 160,000 g, and in this fraction we observed [32P]-labeling of a 96-kDa protein under mono-ADP-ribosylation conditions with [32P]NAD+. The [32P]-labeled protein had the same molecular weight as the EhADH2 monomer. Because of the importance of monoADP-ribosylation in the regulation of many physiological processes, the aim of this study was to determine whether EhADH2 is ADP-ribosylated, and what would be the consequence of this modification on its alcohol and aldehyde dehydrogenase enzymatic activities. This study describes the ADP-ribosylation of EhADH2. This modification did not have an effect on the enzymatic activities, but it may regulate other functions of EhADH2.

Alcohol Dehydrogenase↗
PubMed2001

Recognition of carbohydrate epitopes specific for electron-dense granule antigens from Entamoeba histolytica by monoclonal antibodies in the cecal content of infected hamsters.

The pathogenic trophozoites of Entamoeba histolytica produce and secrete electron-dense granules (EDG) containing collagenase, considered a virulence factor. Two monoclonal antibodies (MAbs) (L7.1 and L1.1) anti-EDG antigens were raised. MAb L7.1 has been reported to recognize proteic EDG antigens and MAb L1.1 reacted with a carbohydrate epitope. These epitopes were present in axenic and xenic amoebas. To detect EDG antigens by a enzyme-linked immunosorbent assay (ELISA) in a experimental model of early intestinal amoebiasis, both MAbs were employed. E. histolytica HM1 axenic and monoxenic trophozoites were inoculated into the cecum according to the washed-closed cecal loop technique. The cecal content was recovered at 8, 24, and 48 h post-inoculation. Antigens from EDG in whole trophozoites and cell-free supernatants were detected. Our results indicate that it is possible to detect EDG antigens in the cecal content of hamsters in the early phase of the invasive amoebiasis.

Animals↗
PubMed2002

Possible role of calcium ions, calcium channels and calmodulin in excystation and metacystic development of Entamoeba invadens.

The effect of calcium ions (Ca(2+)) and calmodulin (CaM) on the excystation and metacystic development of Entamoeba invadens was examined by transfer of cysts to a growth medium containing calcium antagonists and CaM inhibitors. Excystation, which was assessed by counting the number of metacystic amoebae after induction of excystation, was inhibited by the calcium chelators ethyleneglycol bis (beta-aminoethyl ether)- N,N'-tetraacetate (EGTA) and ethylene-diaminetetraacetate (EDTA), with EDTA being more potent than EGTA. The inhibitory effect of higher concentrations of these chelators on excystation was associated with reduced viability of cysts. Metacystic development, when determined by the number of nuclei in an amoeba, was delayed by EGTA, because the percentage of four-nucleate amoebae was higher than in controls at day 3 of incubation. EDTA made metacystic development unusual by producing a large number of metacystic amoebae with more than ten nuclei. The inhibition of excystation by these chelators was partially abrogated by their removal. A putative antagonist of intracellular calcium flux, 8-( N,N-diethylamino) octyl-3,4,5-trimethoxybenzoate (TMB-8) also inhibited the excystation and metacystic development, but had little effect on cyst viability. The slow Na(+)-Ca(2+) channel blocker bepridil but not verapamil inhibited the excystation and metacystic development, associating with reduced cyst viability at higher concentrations. The inhibitory effect of bepridil on excystation was abrogated by removal of the drug. The CaM inhibitor trifIuoperazine (TFP) but not W-7 [ N-(6-aminohexyl)-chloro-1-naphtalene sulphonamide] inhibited the excystation and metacystic development. The inhibitory effect of TFP on excystation was also abrogated by removal of the drug. These results indicate that extracellular calcium ions, amoebic intracellular calcium flux, calcium channels, and a CaM-dependent process contribute to the excystation and metacystic development of E. invadens.

Animals↗