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W A Petri

Publications and source records attributed to W A Petri.

At least 19 recordsLinked to original sources

Zoonotic implications of the swine-transmitted protozoal infections.

Pig production is an important part of the economy in many countries. Domestic and wild pigs (Sus scrofa) are susceptible to a wide range of infectious and parasitic diseases. Some of these diseases are specifically limited to pigs while some of the other diseases are shared with other species of wildlife and domestic livestock. As the numbers and geographic distribution of wild and domestic swines continue to increase, it is certain that the number of contacts between these swines and domestic livestock will also increase, as will the probability of human exposure to the parasites of swine directly or indirectly. Here, we will discuss the protozoal infections of pigs, which have the potential to infect humans and provide reasonable risk assessment for zoonotic transmission.

Animals↗

Entamoeba histolytica activates host cell caspases during contact-dependent cell killing.

Entamoeba histolytica is a human intestinal parasite that causes amoebic colitis as well as liver abscesses. Host tissues are damaged through a three-step process involving adherence, contact-dependent cytolysis, and phagocytosis. These three processes all contribute to the pathogenicity of this parasite. Adherence is provided by the Gal/GalNAc adherence lectin. Host cells are lysed in a contact-dependent fashion. There is evidence that suggests that this contact-dependent killing involves the induction of the host cell's apoptotic machinery. Phagocytosis can then occur, consistent with metazoan apoptotic clearance.

Animals↗

Amebiasis and mucosal IgA antibody against the Entamoeba histolytica adherence lectin in Bangladeshi children.

Amebiasis is the third leading parasitic cause of death worldwide, and it is not known whether immunity is acquired from a previous infection. An investigation was done to determine whether protection from intestinal infection correlated with mucosal or systemic antibody responses to the Entamoeba histolytica GalNAc adherence lectin. E. histolytica colonization was present in 0% (0/64) of children with and 13.4% (33/246) of children without stool IgA anti-GalNAc lectin antibodies (P= .001). Children with stool IgA lectin-specific antibodies at the beginning of the study had 64% fewer new E. histolytica infections by 5 months (3/42 IgA(+) vs. 47/227 IgA(-); P= .03). A stool antilectin IgA response was detected near the time of resolution of infection in 67% (12/18) of closely monitored new infections. It was concluded that a mucosal IgA antilectin antibody response is associated with immune protection against E. histolytica colonization. The demonstration of naturally acquired immunity offers hope for a vaccine to prevent amebiasis.

Animals↗

Identification and characterization of an Entamoeba histolytica upstream regulatory element 3 sequence-specific DNA-binding protein containing EF-hand motifs.

The hgl5 gene of Entamoeba histolytica is negatively regulated through the upstream regulatory element 3 (URE3) DNA motif TATTCTATT. This motif is also present and significant in the function of the E. histolytica fdx gene promoter. A yeast one-hybrid screen was used to identify an E. histolytica cDNA encoding a protein (URE3-BP) that recognized this DNA motif. Analysis of the predicted amino acid sequence demonstrated the presence of two EF-hand motifs but identified no canonical DNA binding motifs. URE3-BP, expressed in bacteria, demonstrated Ca(2+)-dependent and sequence-specific recognition of the URE3 DNA sequence as assessed by electrophoretic mobility shift assays. Antibodies raised against URE3-BP blocked the formation of the URE3 DNA-protein complex by native nuclear extracts. The URE3-BP protein was present in the E. histolytica nucleus and cytoplasm with an apparent molecular mass of 22.6 kDa. Our results represent the first use of a yeast genetic screen to identify, on the basis of function, a DNA-binding protein of an early branching eukaryote. Since the URE3 DNA can modulate gene expression in both a positive and negative manner, this protein may have more than one mechanism of interaction with transcriptional machinery. Characterization of URE3-BP should provide insight into transcription regulation and virulence control in this parasite.

Amino Acid Sequence↗

Identification of two Entamoeba histolytica sequence-specific URE4 enhancer-binding proteins with homology to the RNA-binding motif RRM.

To study transcriptional regulation in the lower branching eukaryote Entamoeba histolytica, we have identified two sequence-specific DNA-binding proteins that recognize the upstream regulatory element URE4, an enhancer that regulates expression of the Gal/GalNAc lectin heavy subunit gene hgl5. A chromatographic purification of E. histolytica nuclear extracts by gel filtration, cation exchange, and sequence-specific DNA affinity chromatography led to a 700-fold increase in URE4 binding activity and the appearance of two dominant protein species with molecular masses of 28 and 18 kDa. These proteins, termed E. histolytica enhancer-binding proteins 1 and 2 (EhEBP1 and EhEBP2), were sequenced by tandem mass spectroscopy and their corresponding cDNA clones identified. Recombinant EhEBP1 and EhEBP2 were able to bind double-stranded oligonucleotides bearing the URE4 motif in a sequence-specific manner, and antibodies raised against EhEBP1 were able to interfere with the formation of URE4-protein complexes in crude nuclear extracts. Overexpression of EhEBP1 in E. histolytica trophozoites resulted in a 7-fold drop in promoter activity in transiently transfected reporter gene constructs when the URE4 motif was present, confirming its ability to specifically recognize the URE4 motif and suggesting that additional cofactors may be required for transcriptional activation by URE4. Further characterization and identification of binding partners for EhEBP1 and EhEBP2, the first proteins with demonstrated sequence-specific DNA binding activity to be identified in E. histolytica, should provide new insights into transcriptional regulation in this protozoan parasite.

Amino Acid Sequence↗

Entamoeba histolytica: genetic diversity of clinical isolates from Bangladesh as demonstrated by polymorphisms in the serine-rich gene.

The varied organ tropisms and clinical presentations of infection by Entamoeba histolytica have stimulated interest in the role of parasite genetic diversity in virulence. We investigated genetic diversity among 54 E. histolytica isolates from Bangladesh by analyzing polymorphism in the serine-rich gene by nested PCR on DNA extracted from stool and liver aspirate pus. We detected both size and restriction site polymorphisms among the isolates within this endemic area. A combination of the nested PCR results and the AluI digestion of the PCR products examined yielded 25 distinct DNA banding patterns among the 42 stool isolates and an additional 9 distinct patterns among the 12 liver abscess isolates. Approximately half of the isolates had unique polymorphisms. Interestingly, the majority of E. histolytica from the liver had polymorphisms which were not present in intestinal isolates from the same geographic area. These data are consistent with the existence of genetic differences between E. histolytica which cause intestinal and those which cause hepatic disease. We conclude that there is genetic diversity within E. histolytica isolates from an endemic population as reflected in serine-rich E. histolytica protein gene polymorphism. The correlation of genetic differences with the pathogenic potential of E. histolytica strains and the implications of genetic diversity for the immunoprophylaxis of amebiasis require further study.

Amino Acid Sequence↗

Emerging and reemerging intestinal protozoa.

The intestinal protozoa have gained importance to physicians practicing medicine in the United States, Canada, and Europe during recent years as a result of increasing world travel, the globalization of the world's economy, and the growing number of chronically immunosuppressed people. During the spring of 1996, Cyclospora cayetanensis caused diarrhea in approximately 1500 people exposed to Guatemalan raspberries. This epidemic recurred in 1997, emphasizing the risks of the global economy and food supply on which we depend. In addition to importation of intestinal protozoa from the tropics, AIDS and the increasing use of organ transplants have created a new population of people at risk for chronic infection by ubiquitous protozoa previously not known to cause serious human disease. These infections include cryptosporidiosis, isosporiasis, and microsporidiosis. Finally, Entamoeba histolytica, the etiologic agent of invasive amebiasis, has only recently been recognized to be a distinct species from a nonpathogenic but indistinguishable (by light microscopy) intestinal commensal, Entamoeba dispar. The rapidly changing epidemiology of these intestinal protozoa, as well as new approaches to diagnosis and treatment of these protozoa, are discussed.

Journal Article↗

Intermediate subunit of the Gal/GalNAc lectin of Entamoeba histolytica is a member of a gene family containing multiple CXXC sequence motifs.

Killing by Entamoeba histolytica requires parasite adherence to host galactose- and N-acetyl-D-galactosamine (Gal/GalNAc)-containing cell surface receptors. A 260-kDa heterodimeric E. histolytica Gal/GalNAc lectin composed of heavy (Hgl) and light (Lgl) subunits has been previously described. Here we present the cloning and characterization of Igl, a 150-kDa intermediate subunit of the Gal/GalNAc lectin. Igl, Hgl, and Lgl colocalized on the surface membrane of trophozoites. Two unlinked copies of genes encoding Igl shared 81% amino acid sequence identity (GenBank accession no. AF337950 and AF337951). They encoded cysteine-rich proteins with amino- and carboxy-terminal hydrophobic signal sequences characteristic of glycosylphosphatidylinositol (GPI)-anchored membrane proteins. The igl genes lacked carbohydrate recognition domains but were members of a large family of amebic genes containing CXXC and CXC motifs. These data indicate that Igl is part of the parasite's multimolecular Gal/GalNAc adhesin required for host interaction.

Amino Acid Motifs↗

Entamoeba histolytica: deletion of the GPI anchor signal sequence on the Gal/GalNAc lectin light subunit prevents its assembly into the lectin heterodimer.

Adherence and cytotoxicity of Entamoeba histolytica require the function of a heterodimeric galactose and N-acetylgalactosamine (Gal/GalNAc)-specific lectin. The lectin heavy subunit (Hgl) contains a carbohydrate recognition domain and mediates inside-out cell signaling via its cytoplasmic tail. The function of the lectin light subunit (Lgl) is unknown. The lectin has a unique mechanism of membrane association: Hgl is transmembrane but Lgl is glycosylphosphatidylinositol (GPI) anchored. The role of the GPI anchor signal sequence in heterodimer assembly was tested. Epitope-tagged Lgl with or without the GPI anchor addition signal was expressed in E. histolytica trophozoites. Tagged Lgl did not assemble with Hgl into a lectin heterodimer in the absence of the GPI addition signal. Consistent with previous results that only the Hgl subunit mediates adherence, the monomeric Lgl without the GPI anchor signal lacked Gal/GalNAc-binding activity.

Animals↗

Amebic liver abscess.

Amebiasis is a widespread parasitic disease caused by Entamoeba histolytica. This protozoan organism is the third leading parasitic cause of death in the developing world and is an important health risk to travelers in endemic areas. Amebiasis most commonly results in asymptomatic colonization of the gastrointestinal tract, but some patients may develop intestinal invasive disease or extraintestinal disease-amebic liver abscess being the most common extraintestinal manifestation. This article reviews epidemiologic features, pathophysiology, clinical features, diagnostic tests, imaging studies, treatment of amebic liver abscess, and prevention measures.

Animals↗

Caspase 3-dependent killing of host cells by the parasite Entamoeba histolytica.

The parasite Entamoeba histolytica is named for its ability to lyse host tissues. To determine the factors responsible, we have initiated an examination of the contribution of parasite virulence factors and host caspases to cellular destruction by the parasite. Amoebic colitis in C3H/HeJ mice was associated with extensive host apoptosis at sites of E. histolytica invasion. In vitro studies of E. histolytica-Jurkat T-cell interactions demonstrated that apoptosis required contact via the amoebic Gal/GalNAc lectin, but was unaffected by 75% inhibition of the amoebic cysteine proteinases. Parasite-induced DNA fragmentation was unaffected in caspase 8-deficient Jurkat cells treated with the caspase 9 inhibitor Ac-LEHD-fmk. In contrast, caspase 3-like activity was observed within minutes of E. histolytica contact and the caspase 3 inhibitor Ac-DEVD-CHO blocked Jurkat T cell death, as measured by both DNA fragmentation and 51Cr release. These data demonstrate rapid parasite-induced activation of caspase 3-like caspases, independent of the upstream caspases 8 and 9, which is required for host cell death.

Animals↗

Protozoan parasites that infect the gastrointestinal tract.

Protozoa that infect the gastrointestinal tract include the deadly parasite Entamoeba histolytica;Giardia lamblia, the most common cause of waterborne disease outbreaks; and the large group of spore-forming parasites that share a green algae symbiont and a predilection for causing chronic diarrhea in immunocompromised persons. Some recent advances in the epidemiology, pathogenesis, diagnosis, and treatment of the diseases caused by these protozoa are discussed in this review.

Journal Article↗

Diagnosis of amebic liver abscess and intestinal infection with the TechLab Entamoeba histolytica II antigen detection and antibody tests.

A noninvasive diagnostic test for amebic liver abscess is needed, because amebic and bacterial abscesses appear identical on ultrasound or computer tomography and because it is rarely possible to identify Entamoeba histolytica in stool specimens from patients with amebic liver abscess. Here we report a method of detection in serum of circulating E. histolytica Gal/GalNAc lectin to diagnose amebic liver abscess, which was used in patients from Dhaka, Bangladesh. The TechLab E. histolytica II test (which differentiates the true pathogen E. histolytica from Entamoeba dispar) detected Gal/GalNAc lectin in the sera of 22 of 23 (96%) amebic liver abscess patients tested prior to treatment with the antiamebic drug metronidazole and 0 of 70 (0%) controls. After 1 week of treatment with metronidazole, 9 of 11 (82%) patients became serum lectin antigen negative. The sensitivity of the E. histolytica II antigen detection test for intestinal infection was also evaluated. Antigen detection identified E. histolytica infection in 50 samples from 1, 164 asymptomatic preschool children aged 2 to 5 years, including 16 of 16 (100%) culture-positive specimens. PCR analysis of stool specimens was used to confirm that most antigen-positive but culture-negative specimens were true-positive: PCR identified parasite DNA in 27 of 34 (79%) of the antigen-positive, culture-negative stool specimens. Antigen detection was a more sensitive test for infection than antilectin antibodies, which were detected in only 76 of 98 (78%) amebic liver abscess patients and in 26 of 50 (52%) patients with intestinal infection. We conclude that the TechLab E. histolytica II kit is a sensitive means to diagnose hepatic and intestinal amebiasis prior to the institution of metronidazole treatment.

Adult↗

Socioeconomic status, clinical features, laboratory and parasitological findings of hepatic amebiasis patients--a hospital based prospective study in Bangladesh.

Socioeconomic status, clinical, laboratory and parasitological features of 31 hospitalized amebic liver abscess (ALA) and 8 amebic hepatitis (AH) patients were studied. Thirty-seven (94.9%) of the total 39 cases were from low socioeconomic class and 2 (5.1%) were from middle class (p<0.001). Sixteen (51.6%) ALA and 5 (62.5%) AH patients were admitted with duration of disease for 3 weeks or more. Twenty-one (67.7%) ALA and 3 (37.5%) AH cases gave no previous history of diarrhea or dysentery. Epigastric pain was the predominant symptoms in 71% patients compared to high fever (19.4%), nausea and vomiting (9.7%). Neutrophilic leukocytosis was found in 9 (29.0%) ALA and 2 (25%) AH cases. Raised alkaline phosphatase was the predominant abnormal liver function test found elevated in 22 (71.0%) ALA and 5 (62.5%) AH cases. Three (7.7%) of the 5 (12.8%) microscopy positive stool samples yielded growth of Entamoeba histolytica in culture. The right lobe was involved in 28 (90.3%) ALA cases; 29 (93.5%) patients had single abscess. Bacterial super infection was observed in 1 (12.5%) abscess, reactive changes in right lung was observed in 6 (19.4%) ALA and 1 (12.5%) AH cases. Ascaris lumbricoides was the predominant associated intestinal parasite.

Adolescent↗