Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0017536 (giardiasis)
1,714 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To define the potential role of complement and antibody in host defense against Giardia lamblia, the effect of human serum on axenically cultured G. lamblia trophozoites was studied. Sera from patients without a history of giardiasis and with no detectable antibody by an indirect immunofluorescence antibody (IFA) assay (IFA titers less than 1:2) killed from 8 to 76% of trophozoites (n = 23 sera). Whereas less than 10% of parasites incubated in phosphate-buffered saline alone were killed, 16 sera killed from 10 to 25% and six sera killed from 25 to 75%. One serum with an anti-G lamblia antibody titer of 1:128 killed greater than 98% of the parasites. The complement dependency of killing was demonstrated by abrogation of the lethal effect when serum was chelated with EDTA (7 mM) or was heat-inactivated (56 degrees C, 30 min), and, in an immunofluorescence assay, by detection of C3 on parasites killed by normal serum. When the classical complement pathway was selectively blocked by using serum chelated with Mg+2 (2 mM)-EGTA (8 mM), or serum congenitally deficient for the second component of complement, there was no killing; thus, killing was dependent on the presence of an intact classical pathway. In each of three sera from donors with negative IFA titers, an absorbable factor specific for G. lamblia, possibly antibody not detected by IFA, was required for classical pathway activation. To determine if alteration of the surface of G. lamblia would render it an activator of the alternative pathway of complement, trophozoites were studied after cell death or after treatment with neuraminidase or trypsin. In MgEGTA-chelated serum, dead trophozoites activated the alternative pathway as determined by consumption of Factor B and deposition of C3 on their surface. In contrast, untreated or enzyme-treated living parasites did not activate the alternative pathway.
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PMID:Susceptibility of Giardia lamblia trophozoites to the lethal effect of human serum. 669 7

Animals are commonly considered to be potential sources for Giardia lamblia infections in humans, but the extent of zoonotic transmission of G. lamblia remains controversial because of inadequate understanding of its epidemiology. A better understanding of the epidemiology of G. lamblia may be facilitated by a more effective means for classifying G. lamblia isolates. To develop a sequence-based classification system, the gene encoding the metabolic enzyme triose phosphate isomerase (tim) was sequenced from a number of G. lamblia isolates of various host origins. Restriction enzymes were identified that can distinguish among isolates without the need for sequencing, simplifying the application of this approach to the epidemiologic investigation of giardiasis. Isolates from a previously reported epidemic of giardiasis were accurately classified by this technique, further verifying its utility for epidemiologic investigation.
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PMID:The molecular epidemiology of Giardia lamblia: a sequence-based approach. 865 3

Giardia lamblia, the protozoan parasite responsible for giardiasis, requires purine salvage from its host for RNA and DNA synthesis. G. lamblia expresses an unusual purine phosphoribosyltransferase with a high specificity for guanine (GPRTase). The enzyme's sequence significantly diverges from those of related enzymes in other organisms. The transition state analogue immucillinGP is a powerful inhibitor of HGXPRTase from malaria [Li, C. M., et al. (1999) Nat. Struct. Biol. 6, 582-587] and is also a 10 nM inhibitor of G. lamblia GPRTase. Cocrystallization of GPRTase with immucillinGP led unexpectedly to a GPRTase.immucillinG binary complex with an open catalytic site loop. Diffusion of ligands into preformed crystals gave a GPRTase.immucillinGP.Mg(2+).pyrophosphate complex in which the open loop is stabilized by crystal contacts. G. lamblia GPRTase exhibits substantial structural differences from known purine phosphoribosyltransferases at positions remote from the catalytic site, but conserves most contacts to the bound inhibitor. The filled catalytic site with an open catalytic loop provides insight into ligand binding. One active site Mg(2+) ion is chelated to pyrophosphate, but the other is chelated to two conserved catalytic site carboxylates, suggesting a role for these amino acids. This arrangement of Mg(2+) and pyrophosphate has not been reported in purine phosphoribosyltransferases. ImmucillinG in the binary complex is anchored by its 9-deazaguanine group, and the iminoribitol is disordered. No Mg(2+) or pyrophosphate is detected; thus, the 5'-phosphoryl group is needed to immobilize the iminoribitol prior to magnesium pyrophosphate binding. Filling the catalytic site involves (1) binding the purine ring, (2) anchoring the 5'-phosphate to fix the ribosyl group, (3) binding the first Mg(2+) to Asp125 and Glu126 carboxyl groups and binding Mg(2+).pyrophosphate, and (4) closing the catalytic site loop and formation of bound (Mg(2+))(2). pyrophosphate prior to catalysis. Guanine specificity is provided by two peptide carbonyl oxygens hydrogen-bonded to the exocyclic amino group and a weak interaction to O6. Transition state formation involves N7 protonation by Asp129 acting as the general acid.
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PMID:Crystal structures of Giardia lamblia guanine phosphoribosyltransferase at 1.75 A(,). 1084 57

An assay that uses heminested PCR-restriction fragment length polymorphism analysis for the detection and genotyping of Giardia duodenalis on the basis of polymorphism in the triose phosphate isomerase (tpi) gene was developed. This assay was evaluated with DNA extracted from purified parasite material, bacterial cultures, whole human feces containing G. duodenalis and other parasites, and their corresponding immunofluorescence-stained fecal smears on glass microscope slides. The assay was specific and discriminated between G. duodenalis assemblages A and B. RFLP analysis further distinguished two groups (designated groups I and II) within assemblage A. Among 35 DNA samples extracted from whole feces from patients with confirmed sporadic giardiasis, the tpi gene was amplified from 33 (94%). Of these, nine (27%) samples contained assemblage A group II, 21 (64%) contained assemblage B, and 3 (9%) contained a mixture of assemblage A group II and assemblage B. The tpi gene of G. duodenalis assemblage B was amplified from 21 of 24 (88%) DNA samples extracted from whole feces from patients with confirmed cases of infection in a nursery outbreak. No amplification was detected from the remaining three DNA samples. Overall, analysis of DNA extracted from material recovered from stained microscope slides identified identical G. duodenalis genotypes in 35 (65%) of the 54 samples for which a genotype was established with DNA from whole feces. The heminested PCR method developed is sensitive, simple, and rapid to perform and is applicable for the analysis of other intestinal pathogens.
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PMID:Sensitive PCR-restriction fragment length polymorphism assay for detection and genotyping of Giardia duodenalis in human feces. 1182 55

The objective of this study was to evaluate the efficacy of a vaccine in the prevention of Giardia duodenalis infection in calves. Six 2-week old calves were vaccinated subcutaneously with a sonicated G. duodenalis trophozoite vaccine. Six 2-week old control calves received a subcutaneous injection of sterile phosphate-buffered-saline mixed with adjuvant. Injections were repeated after 28 days. Eleven days after the second injection, calves were challenged orally with 1x10(5) purified G. duodenalis cysts from a naturally infected calf. Throughout the study, fecal samples were collected at regular intervals and examined for the presence of G. duodenalis cysts. Blood samples were collected weekly until G. duodenalis challenge and bi-weekly following challenge. Calves were euthanized 14 days after challenge and G. duodenalis trophozoites within the small intestines were enumerated. Serum antibody titers were significantly higher in vaccinated compared to non-vaccinated calves. Vaccinated calves tended to excrete more G. duodenalis cysts in their feces than non-vaccinated calves. The number of trophozoites in the small intestine was not different between vaccinated and non-vaccinated calves. Changes consistent of moderate enteritis were found in the intestines of one vaccinated and one non-vaccinated calf. Despite a serological immune response following vaccination, this vaccine was not efficacious in preventing giardiasis or reducing cyst shedding in calves.
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PMID:Efficacy of vaccination in preventing giardiasis in calves. 1735 Jul 65

Giardia duodenalis is a flagellated protist that causes gastrointestinal disease throughout the world. In the Philippines, study on G. duodenalis is limited. It is also believed that prevalence rates of this organism in the country are underestimated. In this study, stool samples from residents living in a slum area in Manila were collected. These were examined under microscopy for identification of common helminthic and protistan parasites. Results showed that 22.05% of 2,354 stool samples collected contained Giardia cysts. A fraction of samples (n = 133) positive for Giardia cysts were set aside. Genomic DNA was extracted from these samples and a polymerase chain reaction-restriction fragment length polymorphism procedure based on the organism's triose phosphate isomerase gene was utilized. This particular procedure is capable of distinguishing assemblages or genotypes within G. duodenalis. The highest identified assemblage was Assemblage B (86.47%). The two genotypes of Assemblage A were also detected. This is the first report on the identification of genotypes of G. duodenalis in the Philippines. The results of this study can serve as basis for future control and prevention of giardiasis and parasitism in the country.
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PMID:Genotyping of Giardia duodenalis isolates among residents of slum area in Manila, Philippines. 1740 23

Investigators tried to correlate clinical presentation of giardiasis to the different genotypes, but controversial data were described through the last decade. The clinical manifestations of 89 Giardia patients were classified into:- GI: 52 symptomatic patients and GII: 37 asymptomatic patients. Genetic characterization of G. lamblia of the patients' fecal samples was performed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) technique b using triose phosphate isomerase gene (tpi). Forty-two patients had genotype A1 (47.19%), 16 had genotype A2 (17.98%), 10 had genotype B (11.23%) and 19 had mixed genotype infection (21.35%). However, PCR-RFLP failed to determine Giardia genotype of only two cases (2.25%). The 20 control fecal samples obtained from healthy subjects showed negative results for G. lamblia by PCR-RFLP method. Of 52 cases in GI, the prevalence genotype A1 was 44.23%, genotype A2 was 19.23%, genotype B was 13.46%, mixed genotype infection was 21.15% and undetermined genotype was 1.92% as compared to 51.35%, 16.21%, 8.11%, 21.62% and 2.70% in GII, respecttively. There was no significant difference between both groups as regard the different Giardia genotypes (p>0.05). Statistical analysis of each symptom in different genotypes revealed insignificant (p>0.05). The results denied any correlation between G. lamblia genotype and the clinical presentation of giardiasis.
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PMID:Genetic characterization of Giardia lamblia isolates from Egyptian patients with relation to clinical giardiasis. 1885 27

Giardial diarrhea in a birth cohort of 452 children in an urban slum in South India was characterized. Of the 155 episodes that occurred in 99 children, 73% were acute diarrhea. Children with better educated mothers and a toilet at home had lower odds of acquiring giardial diarrhea, whereas low socioeconomic status and drinking municipal water were associated with greater risk. Children with co-infections tended to have a slightly longer duration of diarrhea (P = 0.061) and showed significantly more wasting after an episode than children with diarrhea resulting from Giardia alone (P = 0.032). Among the 99 cases, 50 diarrheal and 51 asymptomatic Giardia positive samples were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) at the triose phosphate isomerase gene. Assemblage B was predominant both in giardial diarrhea (80%) and asymptomatic giardiasis (94%). Children with Assemblage A subgroup-II alone or dual infections with both assemblage A and B had diarrhea more frequently (P = 0.07).
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PMID:Giardia duodenalis assemblages associated with diarrhea in children in South India identified by PCR-RFLP. 1914 32

Two major genotypic assemblages of Giardia intestinalis infect humans; the nested real-time polymerase chain reaction (PCR) was used for targeting the triose phosphate isomerase (tpi) gene to detect and genotype G. intestinalis in human feces in Egypt. Among 97 fecal samples, 30 (31%) were diagnosed as giardiasis by saline wet mount microscopy after staining with Lugol's iodine. The tpi gene was amplified from 41 (42.3%) fecal samples, of which 11 were microscopy-negative specimens. Of the total samples, 24 (58.5%) contained assemblage A group I, and 7 (17.1%) were assemblage A group II from the group of patients complaining of intermittent diarrhea. Eight (19.5%) samples contained assemblage B from patients with persistent diarrhea. Two (5%) samples had a mixture of assemblage A group II and assemblage B. The technique was able to detect as few as 20 trophozoites per PCR on fecal DNA-isolated, microscopy-negative, and quantitative (q)PCR-positive specimens; there was a higher average cycle threshold value than microscopy-positive and qPCR-positive specimens, suggesting that they represented true, low-burden infections. In conclusion, we could genotype G. intestinalis from fresh stool samples in Egypt; in infections commonly presented with intermittent diarrhea, the most prevalent genotype was assemblage A group I. The most vulnerable age group included 10- to 20-yr-old individuals.
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PMID:Real-time PCR/RFLP assay to detect Giardia intestinalis genotypes in human isolates with diarrhea in Egypt. 1925 68

Human giardiasis, the gastrointestinal infection caused by two genetically different groups (or assemblages) of Giardia duodenalis, is very common worldwide, and its prevalence is higher in developing countries. However, few surveys in these regions have been performed to include a genetic characterization of the parasite, which is necessary to unravel the complex epidemiology of the infection. In this work, we screened 120 faecal samples collected from Sahrawi children in 2003-2005, and found 41 (34.2%) of them to be positive, using immunofluorescent microscopy, for the presence of G. duodenalis cysts. Molecular characterization of the isolates was performed by RFLP and/or sequence analysis of the triose phosphate isomerase (tpi) and the glutamate dehydrogenase (gdh) genes. The results disclosed an unexpectedly high genetic polymorphism among isolates of both assemblages A and B, and a large percentage of the sequences (50% for the tpi gene, and 90% for the gdh gene) from assemblage B isolates characterized by the presence of overlapping nucleotide peaks at specific positions in the chromatograms, which can be attributed to mixed infections or to allelic sequence heterozygosity of single cysts. Notably, this phenomenon was not observed in sequences from assemblage A isolates. These results suggest that the genetic structure is different in isolates of assemblages A and B.
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PMID:High genetic polymorphism among Giardia duodenalis isolates from Sahrawi children. 1947 74


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